Cathepsin V-like polypeptides

Abstract

The present invention provides novel nucleic acids, novel polypeptide sequences encoded by these nucleic acids and uses thereof.

Description

1. BACKGROUND OF THE INVENTION

1.1 Technical Field

The present invention provides novel polynucleotides and proteins encoded by such polynucleotides, along with uses for these polynucleotides and proteins, for example in therapeutic, diagnostic and research methods.

1.2 Background

Technology aimed at the discovery of protein factors (including e.g., cytokines, such as lymphokines, interferons, CSFs, chemokines, and interleukins) has matured rapidly over the past decade. The now routine hybridization cloning and expression cloning techniques clone novel polynucleotides “directly” in the sense that they rely on information directly related to the discovered protein (i.e., partial DNA/amino acid sequence of the protein in the case of hybridization cloning; activity of the protein in the case of expression cloning). More recent “indirect” cloning techniques such as signal sequence cloning, which isolates DNA sequences based on the presence of a now well-recognized secretory leader sequence motif, as well as various PCR-based or low stringency hybridization-based cloning techniques, have advanced the state of the art by making available large numbers of DNA/amino acid sequences for proteins that are known to have biological activity, for example, by virtue of their secreted nature in the case of leader sequence cloning, by virtue of their cell or tissue source in the case of PCR-based techniques, or by virtue of structural similarity to other genes of known biological activity.

Identified polynucleotide and polypeptide sequences have numerous applications in, for example, diagnostics, forensics, gene mapping; identification of mutations responsible for genetic disorders or other traits, to assess biodiversity, and to produce many other types of data and products dependent on DNA and amino acid sequences.

2. SUMMARY OF THE INVENTION

The compositions of the present invention include novel isolated polypeptides, novel isolated polynucleotides encoding such polypeptides, including recombinant DNA molecules, cloned genes or degenerate variants thereof, especially naturally occurring variants such as allelic variants, antisense polynucleotide molecules, and antibodies that specifically recognize one or more epitopes present on such polypeptides, as well as hybridomas producing such antibodies.

The compositions of the present invention additionally include vectors, including expression vectors, containing the polynucleotides of the invention, cells genetically engineered to contain such polynucleotides and cells genetically engineered to express such polynucleotides.

The present invention relates to a collection or library of at least one novel nucleic acid sequence assembled from expressed sequence tags (ESTs) isolated mainly by sequencing by hybridization (SBM), and in some cases, sequences obtained from one or more public databases. The invention relates also to the proteins encoded by such polynucleotides, along with therapeutic, diagnostic and research utilities for these polynucleotides and proteins. These nucleic acid sequences are designated as SEQ ID NO: 1-948 and are provided in the Sequence Listing. In the nucleic acids provided in the Sequence Listing, A is adenine; C is cytosine; G is guanine; T is thymine; and N is any of the four bases. In the amino acids provided in the Sequence Listing, * corresponds to the stop codon.

The nucleic acid sequences of the present invention also include, nucleic acid sequences that hybridize to the complement of SEQ ID NO: 1-948 under stringent hybridization conditions; nucleic acid sequences which are allelic variants or species homologues of any of the nucleic acid sequences recited above, or nucleic acid sequences that encode a peptide comprising a specific domain or truncation of the peptides encoded by SEQ ID NO: 1-948. A polynucleotide comprising a nucleotide sequence having at least 90% identity to an identifying sequence of SEQ ID NO: 1-948 or a degenerate variant or fragment thereof. The identifying sequence can be 100 base pairs in length.

The nucleic acid sequences of the present invention also include the sequence information from the nucleic acid sequences of SEQ ID NO: 1-948. The sequence information can be a segment of any one of SEQ ID NO: 1-948 that uniquely identifies or represents the sequence information of SEQ ID NO: 1-948.

A collection as used in this application can be a collection of only one polynucleotide. The collection of sequence information or identifying information of each sequence can be provided on a nucleic acid array. In one embodiment, segments of sequence information is provided on a nucleic acid array to detect the polynucleotide that contains the segment. The array can be designed to detect full-match or mismatch to the polynucleotide that contains the segment. The collection can also be provided in a computer-readable format.

This invention also includes the reverse or direct complement of any of the nucleic acid sequences recited above; cloning or expression vectors containing the nucleic acid sequences; and host cells or organisms transformed with these expression vectors. Nucleic acid sequences (or their reverse or direct complements) according to the invention have numerous applications in a variety of techniques known to those skilled in the art of molecular biology, such as use as hybridization probes, use as primers for PCR, use in an array, use in computer-readable media, use in sequencing full-length genes, use for chromosome and gene mapping, use in the recombinant production of protein, and use in the generation of anti-sense DNA or RNA, their chemical analogs and the like.

In a preferred embodiment, the nucleic acid sequences of SEQ ID NO: 1-948 or novel segments or parts of the nucleic acids of the invention are used as primers in expression assays that are well known in the art. In a particularly preferred embodiment, the nucleic acid sequences of SEQ ID NO: 1-948 or novel segments or parts of the nucleic acids provided herein are used in diagnostics for identifying expressed genes or, as well known in the art and exemplified by Vollrath et al., Science 258:52-59 (1992), as expressed sequence tags for physical mapping of the human genome.

The isolated polynucleotides of the invention include, but are not limited to, a polynucleotide comprising any one of the nucleotide sequences set forth in SEQ ID NO: 1-948; a polynucleotide comprising any of the full length protein coding sequences of SEQ ID NO: 1-948; and a polynucleotide comprising any of the nucleotide sequences of the mature protein coding sequences of SEQ ID NO: 1-948. The polynucleotides of the present invention also include, but are not limited to, a polynucleotide that hybridizes under stringent hybridization conditions to (a) the complement of any one of the nucleotide sequences set forth in SEQ ID NO: 1-948; (b) a nucleotide sequence encoding any one of the amino acid sequences set forth in the Sequence Listing; (c) a polynucleotide which is an allelic variant of any polynucleotides recited above; (d) a polynucleotide which encodes a species homolog (e.g. orthologs) of any of the proteins recited above; or (e) a polynucleotide that encodes a polypeptide comprising a specific domain or truncation of any of the polypeptides comprising an amino acid sequence set forth in the Sequence Listing.

The isolated polypeptides of the invention include, but are not limited to, a polypeptide comprising any of the amino acid sequences set forth in the Sequence Listing; or the corresponding full length or mature protein. Polypeptides of the invention also include polypeptides with biological activity that are encoded by (a) any of the polynucleotides having a nucleotide sequence set forth in SEQ ID NO: 1-948; or (b) polynucleotides that hybridize to the complement of the polynucleotides of (a) under stringent hybridization conditions. Biologically or immunologically active variants of any of the polypeptide sequences in the Sequence Listing, and “substantial equivalents” thereof (e.g., with at least about 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98% or 99% amino acid sequence identity) that preferably retain biological activity are also contemplated. The polypeptides of the invention may be wholly or partially chemically synthesized but are preferably produced by recombinant means using the genetically engineered cells (e.g. host cells) of the invention.

The invention also provides compositions comprising a polypeptide of the invention. Polypeptide compositions of the invention may further comprise an acceptable carrier, such as a hydrophilic, e.g., pharmaceutically acceptable, carrier.

The invention also provides host cells transformed or transfected with a polynucleotide of the invention.

The invention also relates to methods for producing a polypeptide of the invention comprising growing a culture of the host cells of the invention in a suitable culture medium under conditions permitting expression of the desired polypeptide, and purifying the polypeptide from the culture or from the host cells. Preferred embodiments include those in which the protein produced by such process is a mature form of the protein.

Polynucleotides according to the invention have numerous applications in a variety of techniques known to those skilled in the art of molecular biology. These techniques include use as hybridization probes, use as oligomers, or primers, for PCR, use for chromosome and gene mapping, use in the recombinant production of protein, and use in generation of anti-sense DNA or RNA, their chemical analogs and the like. For example, when the expression of an mRNA is largely restricted to a particular cell or tissue type, polynucleotides of the invention can be used as hybridization probes to detect the presence of the particular cell or tissue mRNA in a sample using, e.g., in situ hybridization.

In other exemplary embodiments, the polynucleotides are used in diagnostics as expressed sequence tags for identifying expressed genes or, as well known in the art and exemplified by Vollrath et al., Science 258:52-59 (1992), as expressed sequence tags for physical mapping of the human genome.

The polypeptides according to the invention can be used in a variety of conventional procedures and methods that are currently applied to other proteins. For example, a polypeptide of the invention can be used to generate an antibody that specifically binds the polypeptide. Such antibodies, particularly monoclonal antibodies, are useful for detecting or quantitating the polypeptide in tissue. The polypeptides of the invention can also be used as molecular weight markers, and as a food supplement.

Methods are also provided for preventing, treating, or ameliorating a medical condition which comprises the step of administering to a mammalian subject a therapeutically effective amount of a composition comprising a polypeptide of the present invention and a pharmaceutically acceptable carrier.

In particular, the polypeptides and polynucleotides of the invention can be utilized, for example, in methods for the prevention and/or treatment of disorders involving aberrant protein expression or biological activity.

The present invention further relates to methods for detecting the presence of the polynucleotides or polypeptides of the invention in a sample. Such methods can, for example, be utilized as part of prognostic and diagnostic evaluation of disorders as recited herein and for the identification of subjects exhibiting a predisposition to such conditions. The invention provides a method for detecting the polynucleotides of the invention in a sample, comprising contacting the sample with a compound that binds to and forms a complex with the polynucleotide of interest for a period sufficient to form the complex and under conditions sufficient to form a complex and detecting the complex such that if a complex is detected, the polynucleotide of interest is detected. The invention also provides a method for detecting the polypeptides of the invention in a sample comprising contacting the sample with a compound that binds to and forms a complex with the polypeptide under conditions and for a period sufficient to form the complex and detecting the formation of the complex such that if a complex is formed, the polypeptide is detected.

The invention also provides kits comprising polynucleotide probes and/or monoclonal antibodies, and optionally quantitative standards, for carrying out methods of the invention. Furthermore, the invention provides methods for evaluating the efficacy of drugs, and monitoring the progress of patients, involved in clinical trials for the treatment of disorders as recited above.

The invention also provides methods for the identification of compounds that modulate (i.e., increase or decrease) the expression or activity of the polynucleotides and/or polypeptides of the invention. Such methods can be utilized, for example, for the identification of compounds that can ameliorate symptoms of disorders as recited herein. Such methods can include, but are not limited to, assays for identifying compounds and other substances that interact with (e.g., bind to) the polypeptides of the invention. The invention provides a method for identifying a compound that binds to the polypeptides of the invention comprising contacting the compound with a polypeptide of the invention in a cell for a time sufficient to form a polypeptide/compound complex, wherein the complex drives expression of a reporter gene sequence in the cell; and detecting the complex by detecting the reporter gene sequence expression such that if expression of the reporter gene is detected the compound the binds to a polypeptide of the invention is identified.

The methods of the invention also provides methods for treatment which involve the administration of the polynucleotides or polypeptides of the invention to individuals exhibiting symptoms or tendencies. In addition, the invention encompasses methods for treating diseases or disorders as recited herein comprising administering compounds and other substances that modulate the overall activity of the target gene products. Compounds and other substances can effect such modulation either on the level of target gene/protein expression or target protein activity.

The polypeptides of the present invention and the polynucleotides encoding them are also useful for the same functions known to one of skill in the art as the polypeptides and polynucleotides to which they have homology (set forth in Table 2); for which they have a signature region (as set forth in Table 3); or for which they have homology to a gene family (as set forth in Table 4). If no homology is set forth for a sequence, then the polypeptides and polynucleotides of the present invention are useful for a variety of applications, as described herein, including use in arrays for detection.

3. DETAILED DESCRIPTION OF THE INVENTION

3.1 Definitions

It must be noted that as used herein and in the appended claims, the singular forms “a”, “an” and “the” include plural references unless the context clearly dictates otherwise.

The term “active” refers to those forms of the polypeptide which retain the biologic and/or immunologic activities of any naturally occurring polypeptide. According to the invention, the terms “biologically active” or “biological activity” refer to a protein or peptide having structural, regulatory or biochemical functions of a naturally occurring molecule. Likewise “immunologically active” or “immunological activity” refers to the capability of the natural, recombinant or synthetic polypeptide to induce a specific immune response in appropriate animals or cells and to bind with specific antibodies.

The term “activated cells” as used in this application are those cells which are engaged in extracellular or intracellular membrane trafficking, including the export of secretory or enzymatic molecules as part of a normal or disease process.

The terms “complementary” or “complementarity” refer to the natural binding of polynucleotides by base pairing. For example, the sequence 5′-AGT-3′ binds to the complementary sequence 3′-TCA-5′. Complementarity between two single-stranded molecules may be “partial” such that only some of the nucleic acids bind or it may be “complete” such that total complementarity exists between the single stranded molecules. The degree of complementarity between the nucleic acid strands has significant effects on the efficiency and strength of the hybridization between the nucleic acid strands.

The term “embryonic stem cells (ES)” refers to a cell that can give rise to many differentiated cell types in an embryo or an adult, including the germ cells. The term “germ line stem cells (GSCs)” refers to stem cells derived from primordial stem cells that provide a steady and continuous source of germ cells for the production of gametes. The term “primordial germ cells (PGCs)” refers to a small population of cells set aside from other cell lineages particularly from the yolk sac, mesenteries, or gonadal ridges during embryogenesis that have the potential to differentiate into germ cells and other cells. PGCs are the source from which GSCs and ES cells are derived The PGCs, the GSCs and the ES cells are capable of self-renewal. Thus these cells not only populate the germ line and give rise to a plurality of terminally differentiated cells that comprise the adult specialized organs, but are able to regenerate themselves.

The term “expression modulating fragment,” EMF, means a series of nucleotides which modulates the expression of an operably linked ORF or another EMF.

As used herein, a sequence is said to “modulate the expression of an operably linked sequence” when the expression of the sequence is altered by the presence of the EMF. EMFs include, but are not limited to, promoters, and promoter modulating sequences (inducible elements). One class of EMFs are nucleic acid fragments which induce the expression of an operably linked ORF in response to a specific regulatory factor or physiological event.

The terms “nucleotide sequence” or “nucleic acid” or “polynucleotide” or “oligonculeotide” are used interchangeably and refer to a heteropolymer of nucleotides or the sequence of these nucleotides. These phrases also refer to DNA or RNA of genomic or synthetic origin which may be single-stranded or double-stranded and may represent the sense or the antisense strand, to peptide nucleic acid (PNA) or to any DNA-like or RNA-like material. In the sequences herein A is adenine, C is cytosine, T is thymine, G is guanine and N is A, C, G or T (U). It is contemplated that where the polynucleotide is RNA, the T (thymine) in the sequences provided herein is substituted with U (uracil). Generally, nucleic acid segments provided by this invention may be assembled from fragments of the genome and short oligonucleotide linkers, or from a series of oligonucleotides, or from individual nucleotides, to provide a synthetic nucleic acid which is capable of being expressed in a recombinant transcriptional unit comprising regulatory elements derived from a microbial or viral operon, or a eukaryotic gene.

The terms “oligonucleotide fragment” or a “polynucleotide fragment”, “portion,” or “segment” or “probe” or “primer” are used interchangeably and refer to a sequence of nucleotide residues which are at least about 5 nucleotides, more preferably at least about 7 nucleotides, more preferably at least about 9 nucleotides, more preferably at least about 11 nucleotides and most preferably at least about 17 nucleotides. The fragment is preferably less than about 500 nucleotides, preferably less than about 200 nucleotides, more preferably less than about 100 nucleotides, more preferably less than about 50 nucleotides and most preferably less than 30 nucleotides. Preferably the probe is from about 6 nucleotides to about 200 nucleotides, preferably from about 15 to about 50 nucleotides, more preferably from about 17 to 30 nucleotides and most preferably from about 20 to 25 nucleotides. Preferably the fragments can be used in polymerase chain reaction (PCR), various hybridization procedures or microarray procedures to identify or amplify identical or related parts of mRNA or DNA molecules. A fragment or segment may uniquely identify each polynucteotide sequence of the present invention. Preferably the fragment comprises a sequence substantially similar to any one of SEQ ID NOs: 1-948.

Probes may, for example, be used to determine whether specific mRNA molecules are present in a cell or tissue or to isolate similar nucleic acid sequences from chromosomal DNA as described by Walsh et al. (Walsh, P. S. et al., 1992, PCR Methods Appl 1:241-250). They may be labeled by nick translation, Klenow fill-in reaction, PCR, or other methods well known in the art. Probes of the present invention, their preparation and/or labeling are elaborated in Sambrook, J. et al., 1989, Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory, NY; or Ausubel, F. M. et al., 1989, Current Protocols in Molecular Biology, John Wiley & Sons, New York N.Y., both of which are incorporated herein by reference in their entirety.

The nucleic acid sequences of the present invention also include the sequence information from the nucleic acid sequences of SEQ ID NOs: 1-948. The sequence information can be a segment of any one of SEQ ID NOs: 1-948 that uniquely identifies or represents the sequence information of that sequence of SEQ ID NO: 1-948. One such segment can be a twenty-mer nucleic acid sequence because the probability that a twenty-mer is fully matched in the human genome is 1 in 300. In the human genome, there are three billion base pairs in one set of chromosomes. Because 4 20 possible twenty-mers exist, there are 300 times more twenty-mers than there are base pairs in a set of human chromosomes. Using the same analysis, the probability for a seventeen-mer to be fully matched in the human genome is approximately 1 in 5. When these segments are used in arrays for expression studies, fifteen-mer segments can be used. The probability that the fifteen-mer is fully matched in the expressed sequences is also approximately one in five because expressed sequences comprise less than approximately 5% of the entire genome sequence.

Similarly, when using sequence information for detecting a single mismatch, a segment can be a twenty-five mer. The probability that the twenty-five mer would appear in a human genome with a single mismatch is calculated by multiplying the probability for a full match (1÷4 25 ) times the increased probability for mismatch at each nucleotide position (3×25). The probability that an eighteen mer with a single mismatch can be detected in an array for expression studies is approximately one in five. The probability that a twenty-mer with a single mismatch can be detected in a human genome is approximately one in five.

The term “open reading frame,” ORF, means a series of nucleotide triplets coding for amino acids without any termination codons and is a sequence translatable into protein.

The terms “operably linked” or “operably associated” refer to functionally related nucleic acid sequences. For example, a promoter is operably associated or operably linked with a coding sequence if the promoter controls the transcription of the coding sequence. While operably linked nucleic acid sequences can be contiguous and in the same reading friame, certain genetic elements e.g. repressor genes are not contiguously linked to the coding sequence but still control transcription/translation of the coding sequence.

The term “pluripotent” refers to the capability of a cell to differentiate into a number of differentiated cell types that are present in an adult organism. A pluripotent cell is restricted in its differentiation capability in comparison to a totipotent cell.

The terms “polypeptide” or “peptide” or “amino acid sequence” refer to an oligopeptide, peptide, polypeptide or protein sequence or fragment thereof and to naturally occurring or synthetic molecules. A polypeptide “fragment,” “portion,” or “segment” is a stretch of amino acid residues of at least about 5 amino acids, preferably at least about 7 amino acids, more preferably at least about 9 amino acids and most preferably at least about 17 or more amino acids. The peptide preferably is not greater than about 200 amino acids, more preferably less than 150 amino acids and most preferably less than 100 amino acids. Preferably the peptide is from about 5 to about 200 amino acids. To be active, any polypeptide must have sufficient length to display biological and/or immunological activity.

The term “naturally occurring polypeptide” refers to polypeptides produced by cells that have not been genetically engineered and specifically contemplates various polypeptides arising from post-translational modifications of the polypeptide including, but not limited to, acetylation, carboxylation, glycosylation, phosphorylation, lipidation and acylation.

The term “translated protein coding portion” means a sequence which encodes for the full length protein which may include any leader sequence or any processing sequence.

The term “mature protein coding sequence” means a sequence which encodes a peptide or protein without a signal or leader sequence. The “mature protein portion” means that portion of the protein which does not include a signal or leader sequence. The peptide may have been produced by processing in the cell which removes any leader/signal sequence. The mature protein portion may or may not include the initial methionine residue. The methionine residue may be removed from the protein during processing in the cell. The peptide may be produced synthetically or the protein may have been produced using a polynucleotide only encoding for the mature protein coding sequence.

The term “derivative” refers to polypeptides chemically modified by such techniques as ubiquitination, labeling (e.g., with radionuclides or various enzymes), covalent polymer attachment such as pegylation (derivatization with polyethylene glycol) and insertion or substitution by chemical synthesis of amino acids such as omithine, which do not normally occur in human proteins.

The term “variant” (or “analog”) refers to any polypeptide differing from naturally occurring polypeptides by amino acid insertions, deletions, and substitutions, created using, e g.; recombinant DNA techniques. Guidance in determining which amino acid residues may be replaced, added or deleted without abolishing activities of interest, may be found by comparing the sequence of the particular polypeptide with that of homologous peptides and minimizing the number of amino acid sequence changes made in regions of high homology (conserved regions) or by replacing amino acids with consensus sequence.

Alternatively, recombinant variants encoding these same or similar polypeptides may be synthesized or selected by making use of the “redundancy” in the genetic code. Various codon substitutions, such as the silent changes which produce various restriction sites, may be introduced to optimize cloning into a plasmid or viral vector or expression in a particular prokaryotic or eukaryotic system. Mutations in the polynucleotide sequence may be reflected in the polypeptide or domains of other peptides added to the polypeptide to modify the properties of any part of the polypeptide, to change characteristics such as ligand-binding affinities, interchain affinities, or degradation/turnover rate.

Preferably, amino acid “substitutions” are the result of replacing one amino acid with another amino acid having similar structural and/or chemical properties, i.e., conservative amino acid replacements. “Conservative” amino acid substitutions may be made on the basis of similarity in polarity, charge, solubility, hydrophobicity, hydrophilicity, and/or the amphipathic nature of the residues involved. For example, nonpolar (hydrophobic) amino acids include alanine, leucine, isoleucine, valine, proline, phenylalanine, tryptophan, and methionine; polar neutral amino acids include glycine, serine, threonine, cysteine, tyrosine, asparagine, and glutamine; positively charged (basic) amino acids include arginine, lysine, and histidine; and negatively charged (acidic) amino acids include aspartic acid and glutamic acid. “Insertions” or “deletions” are preferably in the range of about 1 to 20 amino acids, more preferably 1 to 10 amino acids. The variation allowed may be experimentally determined by systematically malking insertions, deletions, or substitutions of amino acids in a polypeptide molecule using recombinant DNA techniques and assaying the resulting recombinant variants for activity.

Alternatively, where alteration of function is desired, insertions, deletions or non-conservative alterations can be engineered to produce altered polypeptides. Such alterations can, for example, alter one or more of the biological functions or biochemical characteristics of the polypeptides of the invention. For example, such alterations may change polypeptide characteristics such as ligand-binding affinities, interchain affinities, or degradation/turnover rate. Further, such alterations can be selected so as to generate polypeptides that are better suited for expression, scale up and the like in the host cells chosen for expression. For example, cysteine residues can be deleted or substituted with another amino acid residue in order to eliminate disulfide bridges.

The terms “purified” or “substantially purified” as used herein denotes that the indicated nucleic acid or polypeptide is present in the substantial absence of other biological macromolecules, e.g., polynucleotides, proteins, and the like. In one embodiment, the polynucleotide or polypeptide is purified such that it constitutes at least 95% by weight, more preferably at least 99% by weight, of the indicated biological macromolecules present (but water, buffers, and other small molecules, especially molecules having a molecular weight of less than 1000 daltons, can be present).

The term “isolated” as used herein refers to a nucleic acid or polypeptide separated from at least one other component (e.g., nucleic acid or polypeptide) present with the nucleic acid or polypeptide in its natural source. In one embodiment, the nucleic acid or polypeptide is found in the presence of (if anything) only a solvent, buffer, ion, or other component normally present in a solution of the same. The terms “isolated” and “purified” do not encompass nucleic acids or polypeptides present in their natural source.

The term “recombinant,” when used herein to refer to a polypeptide or protein, means that a polypeptide or protein is derived from recombinant (e.g., microbial, insect, or mammalian) expression systems. “Microbial” refers to recombinant polypeptides or proteins made in bacterial or fungal (e.g., yeast) expression systems. As a product, “recombinant microbial” defines a polypeptide or protein essentially free of native endogenous substances and unaccompanied by associated native glycosylation. Polypeptides or proteins expressed in most bacterial cultures, e.g., E. coli , will be free of glycosylation modifications; polypeptides or proteins expressed in yeast will have a glycosylation pattern in general different from those expressed in mammalian cells.

The term “recombinant expression vehicle or vector” refers to a plasmid or phage or virus or vector, for expressing a polypeptide from a DNA (RNA) sequence. An expression vehicle can comprise a transcriptional unit comprising an assembly of (1) a genetic element or elements having a regulatory role in gene expression, for example, promoters or enhancers, (2) a structural or coding sequence which is transcribed into mRNA and translated into protein, and (3) appropriate transcription initiation and termiination sequences. Structural units intended for use in yeast or eukaryotic expression systems preferably include a leader sequence enabling extracellular secretion of translated protein by a host cell. Alternatively, where recombinant protein is expressed without a leader or transport sequence, it may include an amino terminal methionine residue. This residue may or may not be subsequently cleaved from the expressed recombinant protein to provide a final product.

The term “recombinant expression system” means host cells which have stably integrated a recombinant transcriptional unit into chromosomal DNA or carry the recombinant transcriptional unit extrachromosomally. Recombinant expression systems as defined herein will express heterologous polypeptides or proteins upon induction of the regulatory elements linked to the DNA segment or synthetic gene to be expressed. This term also means host cells which have stably integrated a recombinant genetic element or elements having a regulatory role in gene expression, for example, promoters or enhancers. Recombinant expression systems as defined herein will express polypeptides or proteins endogenous to the cell upon induction of the regulatory elements linked to the endogenous DNA segment or gene to be expressed. The cells can be prokaryotic or eukaryotic.

The term “secreted” includes a protein that is transported across or through a membrane, including transport as a result of signal sequences in its amino acid sequence when it is expressed in a suitable host cell. “Secreted” proteins include without limitation proteins secreted wholly (e.g., soluble proteins) or partially (e.g., receptors) from the cell in which they are expressed. “Secreted” proteins also include without limitation proteins that are transported across the membrane of the endoplasmic reticulum. “Secreted” proteins are also intended to include proteins containing non-typical signal sequences (e.g. Interleukin-1 Beta, see Krasney, P. A. and Young, P. R. (1992) Cytokine 4(2):134-143) and factors released from damaged cells (e.g. Interleukin-1 Receptor Antagonist, see Arend, W. P. et. al. (1998) Annu. Rev. Immunol. 16:27-55)

Where desired, an expression vector may be designed to contain a “signal or leader sequence” which will direct the polypeptide through the membrane of a cell. Such a sequence may be naturally present on the polypeptides of the present invention or provided from heterologous protein sources by recombinant DNA techniques.

The term “stringent” is used to refer to conditions that are commonly understood in the art as stringent. Stringent conditions can include highly stringent conditions (i.e., hybridization to filter-bound DNA in 0.5 M NaHPO 4 , 7% sodium dodecyl sulfate (SDS), 1 mM EDTA at 65° C., and washing in 0.1×SSC/0.1% SDS at 68° C.), and moderately stringent conditions (i.e., washing in 0.2×SSC/0.1% SDS at 42° C.). Other exemplary hybridization conditions are described herein in the examples.

In instances of hybridization of deoxyoligonucleotides, additional exemplary stringent hybridization conditions include washing in 6×SSC/0.05% sodium pyrophosphate at 37° C. (for 14-base oligonucleotides), 48° C. (for 17-base oligos), 55° C. (for 20-base oligonucleotides), and 60° C. (for 23-base oligonucleotides).

As used herein, “substantially equivalent” or “substantially similar” can refer both to nucleotide and amino acid sequences, for example a mutant sequence, that varies from a reference sequence by one or more substitutions, deletions, or additions, the net effect of which does not result in an adverse functional dissimilarity between the reference and subject sequences. Typically, such a substantially equivalent sequence varies from one of those listed herein by no more than about 35% (i.e., the number of individual residue substitutions, additions, and/or deletions in a substantially equivalent sequence, as compared to the corresponding reference sequence, divided by the total number of residues in the substantially equivalent sequence is about 0.35 or less). Such a sequence is said to have 65% sequence identity to the listed sequence. In one embodiment, a substantially equivalent, e.g., mutant, sequence of the invention varies from a listed sequence by no more than 30% (70% sequence identity); in a variation of this embodiment, by no more than 25% (75% sequence identity); and in a further variation of this embodiment, by no more than 20% (80% sequence identity) and in a further variation of this embodiment, by no more than 10% (90% sequence identity) and in a further variation of this embodiment, by no more that 5% (95% sequence identity). Substantially equivalent, e.g., mutant, amino acid sequences according to the invention preferably have at least 80% sequence identity with a listed amino acid sequence, more preferably at least 85% sequence identity, more preferably at least 90% sequence identity, more preferably at least 95% sequence identity, more preferably at least 98% sequence identity, and most preferably at least 99% sequence identity. Substantially equivalent nucleotide sequence of the invention can have lower percent sequence identities, taking into account, for example, the redundancy or degeneracy of the genetic code. Preferably, the nucleotide sequence has at least about 65% identity, more preferably at least about 75% identity, more preferably at least about 80% sequence identity, more preferably at least 85% sequence identity, more preferably at least 90% sequence identity, more preferably at least about 95% sequence identity, more preferably at least 98% sequence identity, and most preferably at least 99% sequence identity. For the purposes of the present invention, sequences having substantially equivalent biological activity and substantially equivalent expression characteristics are considered substantially equivalent. For the purposes of determining equivalence, truncation of the mature sequence (e.g., via a mutation which creates a spurious stop codon) should be disregarded. Sequence identity may be determined, e.g., using the Jotun Hein method (Hein, J. (1990) Methods Enzymol. 183:626-645). Identity between sequences can also be determined by other methods known in the art, e.g. by varying hybridization conditions.

The term “totipotent” refers to the capability of a cell to differentiate into all of the cell types of an adult organism.

The term “transformation” means introducing DNA into a suitable host cell so that the DNA is replicable, either as an extrachromosomal element, or by chromosomal integration. The term “transfection” refers to the taking up of an expression vector by a suitable host cell, whether or not any coding sequences are in fact expressed. The term “infection” refers to the introduction of nucleic acids into a suitable host cell by use of a virus or viral vector.

As used herein, an “uptake modulating fragment,” UMF, means a series of nucleotides which mediate the uptake of a linked DNA fragment into a cell. UMFs can be readily identified using known UMFs as a target sequence or target motif with the computer-based systems described below. The presence and activity of a UMF can be confirmed by attaching the suspected UMF to a marker sequence. The resulting nucleic acid molecule is then incubated with an appropriate host under appropriate conditions and the uptake of the marker sequence is determined. As described above, a UMF will increase the frequency of uptake of a linked marker sequence.

Each of the above terms is meant to encompass all that is described for each, unless the context dictates otherwise.

3.2 Nucleic Acids of the Invention

Nucleotide sequences of the invention are set forth in the Sequence Listing.

The isolated polynucleotides of the invention include a polynucleotide comprising the nucleotide sequences of SEQ ID NO: 1-948; a polynucleotide encoding any one of the peptide sequences of SEQ ID NO: 1-948; and a polynucleotide comprising the nucleotide sequence encoding the mature protein coding sequence of the polynucleotides of any one of SEQ ID NO: 1-948. The polynucleotides of the present invention also include, but are not limited to, a polynucleotide that hybridizes under stringent conditions to (a) the complement of any of the nucleotides sequences of SEQ ID NO: 1-948; (b) nucleotide sequences encoding any one of the amino acid sequences set forth in the Sequence Listing; (c) a polynucleotide which is an allelic variant of any polynucleotide recited above; (d) a polynucleotide which encodes a species homolog of any of the proteins recited above; or (c) a polynucleotide that encodes a polypeptide comprising a specific domain or truncation of the polypeptides of SEQ ID NO: 1-948. Domains of interest may depend on the nature of the encoded polypeptide; e.g., domains in receptor-like polypeptides include ligand-binding, extracellular, transmembrane, or cytoplasmic domains, or combinations thereof; domains in immunoglobulin-like proteins include the variable immunoglobulin-like domains; domains in enzyme-like polypeptides include catalytic and substrate binding domains; and domains in ligand polypeptides include receptor-binding domains.

The polynucleotides of the invention include naturally occurring or wholly or partially synthetic DNA, e.g., cDNA and genomic DNA, and RNA, e.g., mRNA. The polynucleotides may include all of the coding region of the cDNA or may represent a portion of the coding region of the cDNA.

The present invention also provides genes corresponding to the cDNA sequences disclosed herein. The corresponding genes can be isolated in accordance with known methods using the sequence information disclosed herein Such methods include the preparation of probes or primers from the disclosed sequence information for identification and/or amplification of genes in appropriate genomic libraries or other sources of genomic materials. Further 5′ and 3′ sequence can be obtained using methods known in the art. For example, full length cDNA or genomic DNA that corresponds to any of the polynucleotides of SEQ ID NO: 1-948 can be obtained by screening appropriate cDNA or genomic DNA libraries under suitable hybridization conditions using any of the polynucleotides of SEQ ID NO: 1-948 or a portion thereof as a probe. Alternatively, the polynucleotides of SEQ ID NO: 1-948 may be used as the basis for suitable primer(s) that allow identification and/or amplification of genes in appropriate genomic DNA or cDNA libraries.

The nucleic acid sequences of the invention can be assembled from ESTs and sequences (including cDNA and genomic sequences) obtained from one or more public databases, such as dbEST, gbpri, and UniGene. The EST sequences can provide identifying sequence information, representative fragment or segment information, or novel segment information for the full-length gene.

The polynucleotides of the invention also provide polynucleotides including nucleotide sequences that are substantially equivalent to the polynucleotides recited above. Polynucleotides according to the invention can have, e.g., at least about 65%, at least about 70%, at least about 75%, at least about 80%, 81%, 82%, 83%, 84%, more typically at least about 85%, 86%, 87%, 88%, 89%, more typically at least about 90%, 91%, 92%, 93%, 94%, and even more typically at least about 95%, 96%, 97%, 98%, 99% sequence identity to a polynucleotide recited above.

Included within the scope of the nucleic acid sequences of the invention are nucleic acid sequence fragments that hybridize under stringent conditions to any of the nucleotide sequences of SEQ ID NO: 1-948, or complements thereof, which fragment is greater than about 5 nucleotides, preferably 7 nucleotides, more preferably greater than 9 nucleotides and most preferably greater than 17 nucleotides. Fragments of, e.g. 15, 17, or 20 nucleotides or more that are selective for (i.e. specifically hybridize to any one of the polynucleotides of the invention) are contemplated. Probes capable of specifically hybridizing to a polynucleotide can differentiate polynucleotide sequences of the invention from other polynucleotide sequences in the same family of genes or can differentiate human genes from genes of other species, and are preferably based on unique nucleotide sequences.

The sequences falling within the scope of the present invention are not limited to these specific sequences, but also include allelic and species variations thereof. Allelic and species variations can be routinely determined by comparing the sequence provided in SEQ ID NO: 1-948, a representative fragment thereof, or a nucleotide sequence at least 90% identical, preferably 95% identical, to SEQ ID NOs: 1-948 with a sequence from another isolate of the same species. Furthermore, to accommodate codon variability, the invention includes nucleic acid molecules coding for the same amino acid sequences as do the specific ORFs disclosed herein. In other words, in the coding region of an ORF, substitution of one codon for another codon that encodes the same amino acid is expressly contemplated.

The nearest neighbor or homology result for the nucleic acids of the present invention, including SEQ ID NOs: 1-948, can be obtained by searching a database using an algorithm or a program. Preferably, a BLAST which stands for Basic Local Alignment Search Tool is used to search for local sequence alignments (Altshul, S. F. J Mol. Evol. 36 290-300 (1993) and Altschul S. F. et al. J. Mol. Biol. 21:403-410 (1990)). Alternatively a FASTA version 3 search against Genpept, using Fastxy algorithm.

Species homologs (or orthologs) of the disclosed polynucleotides and proteins are also provided by the present invention. Species homologs may be isolated and identified by making suitable probes or primers from the sequences provided herein and screening a suitable nucleic acid source from the desired species.

The invention also encompasses allelic variants of the disclosed polynucleotides or proteins; that is, naturally-occurring alternative forms of the isolated polynucleotide which also encode proteins which are identical, homologous or related to that encoded by the polynucleotides.

The nucleic acid sequences of the invention are further directed to sequences which encode variants of the described nucleic acids. These amino acid sequence variants may be prepared by methods known in the art by introducing appropriate nucleotide changes into a native or variant polynucleotide. There are two variables in the construction of amino acid sequence variants: the location of the mutation and the nature of the mutation. Nucleic acids encoding the amino acid sequence variants are preferably constructed by mutating the polynucleotide to encode an amino acid sequence that does not occur in nature. These nucleic acid alterations can be made at sites that differ in the nucleic acids from different species (variable positions) or in highly conserved regions (constant regions). Sites at such locations will typically be modified in series, e.g., by substituting first with conservative choices (e.g., hydrophobic amino acid to a different hydrophobic amino acid) and then with more distant choices (e.g., hydrophobic amino acid to a charged amino acid), and then deletions or insertions may be made at the target site. Amino acid sequence deletions generally range from about 1 to 30 residues, preferably about 1 to 10 residues, and are typically contiguous. Amino acid insertions include amino- and/or carboxyl-terminal fusions ranging in length from one to one hundred or more residues, as well as intrasequence insertions of single or multiple amino acid residues. Intrasequence insertions may range generally from about 1 to 10 amino residues, preferably from 1 to 5 residues. Examples of terminal insertions include the heterologous signal sequences necessary for secretion or for intracellular targeting in different host cells and sequences such as FLAG or poly-histidine sequences useful for purifying the expressed protein.

In a preferred method, polynucleotides encoding the novel amino acid sequences are changed via site-directed mutagenesis. This method uses oligonucleotide sequences to alter a polynucleotide to encode the desired amino acid variant, as well as sufficient adjacent nucleotides on both sides of the changed amino acid to form a stable duplex on either side of the site of being changed. In general, the techniques of site-directed mutagenesis are well known to those of skill in the art and this technique is exemplified by publications such as, Edelman et al., DNA 2:183 (1983). A versatile and efficient method for producing site-specific changes in a polynucleotide sequence was published by Zoller and Smith, Nucleic Acids Res . 10:6487-6500 (1982). PCR may also be used to create amino acid sequence variants of the novel nucleic acids. When small amounts of template DNA are used as starting material, primer(s) that differs slightly in sequence from the corresponding region in the template DNA can generate the desired amino acid variant. PCR amplification results in a population of product DNA fragments that differ from the polynucleotide template encoding the polypeptide at the position specified by the primer. The product DNA fragments replace the corresponding region in the plasmid and this gives a polynucleotide encoding the desired amino acid variant.

A further technique for generating amino acid variants is the cassette mutagenesis technique described in Wells et al., Gene 34:315 (1985); and other mutagenesis techniques well known in the art, such as, for example, the techniques in Sambrook et al., supra, and Current Protocols in Molecular Biology , Ausubel et al. Due to the inherent degeneracy of the genetic code, other DNA sequences which encode substantially the same or a functionally equivalent amino acid sequence may be used in the practice of the invention for the cloning and expression of these novel nucleic acids. Such DNA sequences include those which are capable of hybridizing to the appropriate novel nucleic acid sequence under stringent conditions.

Polynucleotides encoding preferred polypeptide truncations of the invention can be used to generate polynucleotides encoding chimeric or fusion proteins comprising one or more domains of the invention and heterologous protein sequences.

The polynucleotides of the invention additionally include the complement of any of the polynucleotides recited above. The polynucleotide can be DNA (genomic, cDNA, amplified, or synthetic) or RNA. Methods and algorithms for obtaining such polynucleotides are well known to those of skill in the art and can include, for example, methods for determining hybridization conditions that can routinely isolate polynucleotides of the desired sequence identities.

In accordance with the invention, polynucleotide sequences comprising the mature protein coding sequences corresponding to any one of SEQ ID NO: 1-948, or functional equivalents thereof, may be used to generate recombinant DNA molecules that direct the expression of that nucleic acid, or a functional equivalent thereof, in appropriate host cells. Also included are the cDNA inserts of any of the clones identified herein.

A polynucleotide according to the invention can be joined to any of a variety of other nucleotide sequences by well-established recombinant DNA techniques (see Sambrook J et al. (1989) Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory, NY). Useful nucleotide sequences for joining to polynucleotides include an assortment of vectors, e.g., plasmids, cosmids, lambda phage derivatives, phagemids, and the like, that are well known in the art. Accordingly, the invention also provides a vector including a polynucleotide of the invention and a host cell containing the polynucleotide. In general, the vector contains an origin of replication functional in at least one organism, convenient restriction endonuclease sites, and a selectable marker for the host cell. Vectors according to the invention include expression vectors, replication-vectors, probe generation vectors, and sequencing vectors. A host cell according to the invention can be a prokaryotic or eukaryotic cell and can be a unicellular organism or part of a multicellular organism.

The present invention further provides recombinant constructs comprising a nucleic acid having any of the nucleotide sequences of SEQ ID NOs: 1-948 or a fragment thereof or any other polynucleotides of the invention. In one embodiment, the recombinant constructs of the present invention comprise a vector, such as a plasmid or viral vector, into which a nucleic acid having any of the nucleotide sequences of SEQ ID NOs: 1-948 or a fragment thereof is inserted, in a forward or reverse orientation. In the case of a vector comprising one of the ORFs of the present invention, the vector may further comprise regulatory sequences, including for example, a promoter, operably linked to the ORF. Large numbers of suitable vectors and promoters are known to those of skill in the art and are commercially available for generating the recombinant constructs of the present invention. The following vectors are provided by way of example. Bacterial: pBs, phagescript, PsiX174, pBluescript SK, pBs KS, pNH8a, pNH16a, pNH18a, pNH46a (Stratagene); pTrc99A, pKK223-3, pKK233-3, pDR540, pRIT5 (Pharmacia). Eukaryotic: pWLneo, pSV2cat, pOG44, PXTI, pSG (Stratagene) pSVK3, pBPV, pMSG, pSVL (Pharmacia).

The isolated polynucleotide of the invention may be operably linked to an expression control sequence such as the pMT2 or pED expression vectors disclosed in Kaufmnan et al., Nucleic Acids Res . 19, 4485-4490 (1991), in order to produce the protein recombinantly. Many suitable expression control sequences are known in the art. General methods of expressing recombinant proteins are also known and are exemplified in R. Kaufman, Methods in Enzymology 185, 537-566 (1990). As defined herein “operably linked” means that the isolated polynucleotide of the invention and an expression control sequence are situated within a vector or cell in such a way that the protein is expressed by a host cell which has been transformed (transfected) with the ligated polynucleotide/expression control sequence.

Promoter regions can be selected from any desired gene using CAT (chloramphenicol transferase) vectors or other vectors with selectable markers. Two appropriate vectors are pKK232-8 and pCM7. Particular named bacterial promoters include lac, lacZ, T3, T7, gpt, lambda PR, and trc. Eukaryotic promoters include CMV immediate early, HSV thymidine kinase, early and late SV40, LTRs from retrovirus, and mouse metallothionein-I. Selection of the appropriate vector and promoter is well within the level of ordinary skill in the art. Generally, recombinant expression vectors will include origins of replication and selectable markers permitting transformation of the host cell, e.g., the ampicillin resistance gene of E. coli and S. cerevisiae TRP 1 gene, and a promoter derived from a highly-expressed gene to direct transcription of a downstream structural sequence. Such promoters can be derived from operons encoding glycolytic enzymes such as 3-phosphoglycerate kinase (PGK), a-factor, acid phosphatase, or heat shock proteins, among others. The heterologous structural sequence is assembled in appropriate phase with translation initiation and termination sequences, and preferably, a leader sequence capable of directing secretion of translated protein into the periplasmic space or extracellular medium. Optionally, the heterologous sequence can encode a fusion protein including an amino terminal identification peptide imparting desired characteristics, e.g., stabilization or simplified purification of expressed recombinant product. Useful expression vectors for bacterial use are constructed by inserting a structural DNA sequence encoding a desired protein together with suitable translation initiation and termination signals in operable reading phase with a functional promoter. The vector will comprise one or more phenotypic selectable markers and an origin of replication to ensure maintenance of the vector and to, if desirable, provide amplification within the host. Suitable prokaryotic hosts for transformation include E. coli, Bacillus subtilis, Salmonella typhimurium and various species within the genera Pseudomonas, Streptomyces, and Staphylococcus, although others may also be employed as a matter of choice.

As a representative but non-limiting example, usefull expression vectors for bacterial use can comprise a selectable marker and bacterial origin of replication derived from commercially available plasmids comprising genetic elements of the well known cloning vector pBR322 (ATCC 37017). Such commercial vectors include, for example, pKK223-3 (Pharmacia Fine Chemicals, Uppsala, Sweden) and GEM 1 (Promega Biotech, Madison, Wis., USA). These pBR322 “backbone” sections are combined with an appropriate promoter and the structural sequence to be expressed. Following transformation of a suitable host strain and growth of the host strain to an appropriate cell density, the selected promoter is induced or derepressed by appropriate means (e.g., temperature shift or chemical induction) and cells are cultured for an additional period. Cells are typically harvested by centrimugation, disrupted by physical or chemical means, and the resulting crude extract retained for further purification.

Polynucleotides of the invention can also be used to induce immune responses. For example, as described in Fan et al., Nat. Biotech . 17:870-872 (1999), incorporated herein by reference, nucleic acid sequences encoding a polypeptide may be used to generate antibodies against the encoded polypeptide following topical administration of naked plasmid DNA or following injection, and preferably intra-muscular injection of the DNA. The nucleic acid sequences are preferably inserted in a recombinant expression vector and may be in the form of naked DNA.

3.3 Antisense

Another aspect of the invention pertains to isolated antisense nucleic acid molecules that are hybridizable to or complementary to the nucleic acid molecule comprising the nucleotide sequence of SEQ ID NO: 1-948, or fragments, analogs or derivatives thereof. An “antisense” nucleic acid comprises a nucleotide sequence that is complementary to a “sense” nucleic acid encoding a protein, e.g., complementary to the coding strand of a double-stranded cDNA molecule or complementary to an mRNA sequence. In specific aspects, antisense nucleic acid molecules are provided that comprise a sequence complementary to at least about 10, 25, 50, 100, 250 or 500 nucleotides or an entire coding strand, or to only a portion thereof. Nucleic acid molecules encoding fragments, homologs, derivatives and analogs of a protein of any of SEQ ID NO: 1-948 or antisense nucleic acids complementary to a nucleic acid sequence of SEQ ID NO: 1-948 are additionally provided.

In one embodiment, an antisense nucleic acid molecule is antisense to a “coding region” of the coding strand of a nucleotide sequence of the invention. The term “coding region” refers to the region of the nucleotide sequence comprising codons which are translated into amino acid residues. In another embodiment, the antisense nucleic acid molecule is antisense to a “noncoding region” of the coding strand of a nucleotide sequence of the invention. The term “noncoding region” refers to 5′ and 3′ sequences that flank the coding region that are not translated into amino acids (i.e., also referred to as 5′ and 3′ untranslated regions).

Given the coding strand sequences encoding a nucleic acid disclosed herein (e.g., SEQ ID NO: 1-948, antisense nucleic acids of the invention can be designed according to the rules of Watson and Crick or Hoogsteen base pairing. The antisense nucleic acid molecule can be complementary to the entire coding region of an mRNA, but more preferably is an oligonucleotide that is antisense to only a portion of the coding or noncoding region of an mRNA. For example, the antisense oligonucleotide can be complementary to the region surrounding the translation start site of an mRNA. An antisense oligonucleotide can be, for example, about 5, 10, 15, 20, 25, 30, 35, 40, 45 or 50 nucleotides in length. An antisense nucleic acid of the invention can be constructed using chemical synthesis or enzymatic ligation reactions using procedures known in the art. For example, an antisense nucleicacid (e.g., an antisense oligonucleotide) can be chemically synthesized using naturally occurring nucleotides or variously modified nucleotides designed to increase the biological stability of the molecules or to increase the physical stability of the duplex formed between the antisense and sense nucleic acids, e.g., phosphorothioate derivatives and acridine substituted nucleotides can be used.

Examples of modified nucleotides that can be used to generate the antisense nucleic acid include: 5-fluorouracil, 5-bromouracil, 5-chlorouracil, 5-iodouracil, hypoxanthine, xanthine, 4-acetylcytosine, 5-(carboxyhydroxylmethyl) uracil, 5-carboxymethylaminomethyl-2-thiouridine, 5-carboxymethylaminomethyluracil, dihydrouracil, beta-D-galactosylqueosine, inosine, N6-isopentenyladenine, 1-methylguanine, 1-methylinosine, 2,2-dimethylguanine, 2-methyladenine, 2-methylguanine, 3-methylcytosine, 5-methylcytosine, N6-adenine, 7-methylguanine, 5-methylaminomethyluracil, 5-methoxyaminomethyl-2-thiouracil, beta-D-mannosylqueosine, 5′-methoxycarboxymethyluracil, 5-methoxyuracil, 2-methylthio-N6-isopentenyladenine, uracil-5-oxyacetic acid (v), wybutoxosine, pseudouracil, queosine, 2-thiocytosine, 5-methyl-2-thiouracil, 2-thiouracil, 4-thiouracil, 5-methyluracil, uracil-5-oxyacetic acid methylester, uracil-5-oxyacetic acid (v), 5-methyl-2-thiouracil, 3-(3-amino-3-N-2-carboxypropyl)uracil, (acp3)w, and 2,6-diaminopurine. Alternatively, the antisense nucleic acid can be produced biologically using an expression vector into which a nucleic acid has been subcloned in an antisense orientation (i.e., RNA transcribed from the inserted nucleic acid will be of an antisense orientation to a target nucleic acid of interest, described further in the following subsection).

The antisense nucleic acid molecules of the invention are typically administered to a subject or generated in situ such that they hybridize with or bind to cellular mRNA and/or genomic DNA encoding a protein according to the invention to thereby inhibit expression of the protein, e.g., by inhibiting transcription and/or translation. The hybridization can be by conventional nucleotide complementarity to form a stable duplex, or, for example, in the case of an antisense nucleic acid molecule that binds to DNA duplexes, through specific interactions in the major groove of the double helix. An example of a route of administration of antisense nucleic acid molecules of the invention includes direct injection at a tissue site. Alternatively, antisense nucleic acid molecules can be modified to target selected cells and then administered systemically. For example, for systemic administration, antisense molecules can be modified such that they specifically bind to receptors or antigens expressed on a selected cell surface, e.g., by linking the antisense nucleic acid molecules to peptides or antibodies that bind to cell surface receptors or antigens. The antisense nucleic acid molecules can also be delivered to cells using the vectors described herein. To achieve sufficient intracellular concentrations of antisense molecules, vector constructs in which the antisense nucleic acid molecule is placed under the control of a strong pol II or pol III promoter are preferred.

In yet another embodiment, the antisense nucleic acid molecule of the invention is an α-anomeric nucleic acid molecule. An α-anomeric nucleic acid molecule forms specific double-stranded hybrids with complementary RNA in which, contrary to the usual α-units, the strands run parallel to each other (Gaultier et al. (1987) Nucleic Acids Res 15: 6625-6641). The antisense nucleic acid molecule can also comprise a 2′-o-methylribonucleotide (Inoue et al. (1987) Nucleic Acids Res 15: 6131-6148) or a chimeric RNA-DNA analogue (Inoue et al. (1 987) FEBS Lett 215: 327-330).

3.4 Ribozymes and PNA Moieties

In still another embodiment, an antisense nucleic acid of the invention is a ribozyme. Ribozymes are catalytic RNA molecules with ribonuclease activity that are capable of cleaving a single-stranded nucleic acid, such as an mRNA, to which they have a complementary region. Thus, ribozymes (e.g., hammerhead ribozymes (described in Haselhoff and Gerlach (1988) Nature 334:585-591)) can be used to catalytically cleave mRNA transcripts to thereby inhibit translation of an mRNA. A ribozyme having specificity for a nucleic acid of the invention can be designed based upon the nucleotide sequence of a DNA disclosed herein (ie., SEQ ID NO: 1-948). For example, a derivative of Tetrahymena L-19 IVS RNA can be constructed in which the nucleotide sequence of the active site is complementary to the nucleotide sequence to be cleaved in a SECX-encoding mRNA. See, e.g., Cech et al. U.S. Pat. No. 4,987,071; and Cech et al. U.S. Pat. No. 5,116,742. Alternatively, SECX mRNA can be used to select a catalytic RNA having a specific ribonuclease activity from a pool of RNA molecules. See, e.g., Bartel et al., (1993) Science 261:1411-1418.

Alternatively, gene expression can be inhibited by targeting nucleotide sequences complementary to the regulatory region (e.g., promoter and/or enhancers) to form triple helical structures that prevent transcription of the gene in target cells. See generally, Helene. (1991) Anticancer Drug Des . 6: 569-84; Helene. et al. (1992) Ann. N.Y. Acad. Sci . 660:27-36; and Maher (1992) Bioassays 14: 807-15.

In various embodiments, the nucleic acids of the invention can be modified at the base moiety, sugar moiety or phosphate backbone to improve, e.g., the stability, hybridization, or solubility of the molecule. For example, the deoxyribose phosphate backbone of the nucleic acids can be modified to generate peptide nucleic acids (see Hyrup et al. (1996) Bioorg Med Chem 4: 5-23). As used herein, the terms “peptide nucleic acids” or “PNAs” refer to nucleic acid mimics, e.g., DNA mimics, in which the deoxyribose phosphate backbone is replaced by a pseudopeptide backbone and only the four natural nucleobases are retained. The neutral backbone of PNAs has been shown to allow for specific hybridization to DNA and RNA under conditions of low ionic strength. The synthesis of PNA oligomers can be performed using standard solid phase peptide synthesis protocols as described in Hyrup et al. (1996) above; Perry-O'Keefe et al. (1996) PNAS 93: 14670-675.

PNAs of the invention can be used in therapeutic and diagnostic applications. For example, PNAs can be used as antisense or antigene agents for sequence-specific modulation of gene expression by, e.g., inducing transcription or translation arrest or inhibiting replication. PNAs of the invention can also be used, e.g., in the analysis of single base pair mutations in a gene by, e.g., PNA directed PCR clamping; as artificial irestriction enzymes when used in combination with other enzymes, e.g., S1 nucleases (Hyrup B. (1996) above); or as probes or primers for DNA sequence and hybridization (Hyrup et al. (1996), above; Perry-O'Keefe (1996), above).

In another embodiment, PNAs of the invention can be modified, e.g., to enhance their stability or cellular uptake, by attaching lipophilic or other helper groups to PNA, by the formation of PNA-DNA chimeras, or by the use of liposomes or other techniques of drug delivery known in the art. For example, PNA-DNA chimeras can be generated that may combine the advantageous properties of PNA and DNA. Such chimeras allow DNA recognition enzymes, e.g., RNase H and DNA polymerases, to interact with the DNA portion while the PNA portion would provide high binding affinity and specificity. PNA-DNA chimeras can be linked using linkers of appropriate lengths selected in terms of base stacking, number of bonds between the nucleobases, and orientation (Hyrup (1996) above). The synthesis of PNA-DNA chimeras can be performed as described in Hyrup (1996) above and Finn et al. (1996) Nucl Acids Res 24: 3357-63. For example, a DNA chain can be synthesized on a solid support using standard phosphoramidite coupling chemistry, and modified nucleoside analogs, e.g., 5′(4-methoxytrityl)amino-5′-deoxy-thymidine phosphoramidite, can be used between the PNA and the 5′ end of DNA (Mag et al. (1989) Nucl Acid Res 17: 5973-88). PNA monomers are then coupled in a stepwise manner to produce a chimeric molecule with a 5′ PNA segment and a 3′ DNA segment (Finn et al. (1996) above). Alternatively, chimeric molecules can be synthesized with a 5′ DNA segment and a 3′ PNA segment. See, Petersen et al. (1975) Bioorg Med Chem Lett 5: 1119-11124.

In other embodiments, the oligonucleotide may include other appended groups such as peptides (e.g., for targeting host cell receptors in vivo), or agents facilitating transport across the cell membrane (see, e.g., Letsinger et al., 1989 , Proc. Natl. Acad. Sci. U.S.A . 86:6553-6556; Lemaitre et al., 1987, Proc. Natl. Acad. Sci . 84:648-652; PCT Publication No. W088/09810) or the blood-brain barrier (see, e.g., PCT Publication No. W089/10134). In addition, oligonucleotides can be modified with hybridization triggered cleavage agents (See, e.g., Krol et al., 1988 , BioTechniques 6:958-976) or intercalating agents. (See, e.g., Zon, 1988 , Pharm. Res . 5: 539-549). To this end, the oligonucleotide may be conjugated to another molecule, e.g., a peptide, a hybridization triggered cross-linking agent, a transport agent, a hybridization-triggered cleavage agent, etc.

3.5 Hosts

The present invention further provides host cells genetically engineered to contain the polynucleotides of the invention. For example, such host cells may contain nucleic acids of the invention introduced into the host cell using known transformation, transfection or infection methods. The present invention still further provides host cells genetically engineered to express the polynucleotides of the invention, wherein such polynucleotides are in operative association with a regulatory sequence heterologous to the host cell which drives expression of the polynucleotides in the cell.

Knowledge of nucleic acid sequences allows for modification of cells to permit, or increase, expression of endogenous polypeptide. Cells can be modified (e.g., by homologous recombination) to provide increased polypeptide expression by replacing, in whole or in part, the naturally occurring promoter with all or part of a heterologous promoter so that the cells express the polypeptide at higher levels. The heterologous promoter is inserted in such a manner that it is operatively linked to the encoding sequences. See, for example, PCT International Publication No. WO94/12650, PCT International Publication No. WO92/20808, and PCT International Publication No. WO91/09955. It is also contemplated that, in addition to heterologous promoter DNA, amplifiable marker DNA (e.g., ada, dhfr, and the multifunctional CAD gene which encodes carbamyl phosphate synthase, aspartate transcarbamylase, and dihydroorotase) and/or intron DNA may be inserted along with the heterologous promoter DNA. If linked to the coding sequence, amplification of the marker DNA by standard selection methods results in co-amplification of the desired protein coding sequences in the cells.

The host cell can be a higher eukaryotic host cell, such as a mammalian cell, a lower eukaryotic host cell, such as a yeast cell, or the host cell can be a prokaryotic cell, such as a bacterial cell. Introduction of the recombinant construct into the host cell can be effected by calcium phosphate transfection, DEAE, dextran mediated transfection, or electroporation Davis, L. et al., Basic Methods in Molecular Biology (1986)). The host cells containing one of the polynucleotides of the invention, can be used in conventional manners to produce the gene product encoded by the isolated fragment (in the case of an ORF) or can be used to produce a heterologous protein under the control of the EMF.

Any host/vector system can be used to express one or more of the ORFs of the present invention. These include, but are not limited to, eukaryotic hosts such as HeLa cells, Cv-1 cell, COS cells, 293 cells, and Sf9 cells, as well as prokaryotic host such as E. coli and B. subtilis . The most preferred cells are those which do not normally express the particular polypeptide or protein or which expresses the polypeptide or protein at low natural level. Mature proteins can be expressed in mammalian cells, yeast, bacteria, or other cells under the control of appropriate promoters. Cell-free translation systems can also be employed to produce such proteins using RNAs derived from the DNA constructs of the present invention. Appropriate cloning and expression vectors for use with prokaryotic and eukaryotic hosts are described by Sambrook, et al., in Molecular Cloning: A Laboratory Manual, Second Edition, Cold Spring Harbor, N.Y. (1989), the disclosure of which is hereby incorporated by reference.

Various mammalian cell culture systems can also be employed to express recombinant protein. Examples of mammalian expression systems include the COS-7 lines of monkey kidney fibroblasts, described by Gluzman, Cell 23:175 (1981). Other cell lines capable of expressing a compatible vector are, for example, the C127, monkey COS cells, Chinese Hamster Ovary (CHO) cells, human kidney 293 cells, human epidermal A431 cells, human Colo205 cells, 3T3 cells, CV-1 cells, other transformed primate cell lines, normal diploid cells, cell strains derived from in vitro culture of primary tissue, primary explants, HeLa cells, mouse L cells, BHK, HL60, U937, HaK or Jurkat cells. Mammalian expression vectors will comprise an origin of replication, a suitable promoter and also any necessary ribosome binding sites, polyadenylation site, splice donor and acceptor sites, transcriptional termination sequences, and 5′ flanking nontranscribed sequences. DNA sequences derived from the SV40 viral genome, for example, SV40 origin, early promoter, enhancer, splice, and polyadenylation sites may be used to provide the required nontranscribed genetic elements. Recombinant polypeptides and proteins produced in bacterial culture are usually isolated by initial extraction from cell pellets, followed by one or more salting-out, aqueous ion exchange or size exclusion chromatography steps. Protein refolding steps can be used, as necessary, in completing configuration of the mature protein. Finally, high performance liquid chromatography (HPLC) can be employed for fmal purification steps. Microbial cells employed in expression of proteins can be disrupted by any convenient method, including freeze-thaw cycling, sonication, mechanical disruption, or use of cell lysing agents.

Alternatively, it may be possible to produce the protein in lower eukaryotes such as yeast or insects or in prokaryotes such as bacteria. Potentially suitable yeast strains include Saccharomyces cerevisiae, Schizosaccharomyces pombe , Kluyveromyces strains, Candida, or any yeast strain capable of expressing heterologous proteins. Potentially suitable bacterial strains include Escherichia coli, Bacillus subtilis, Salmonella typhimurium , or any bacterial strain capable of expressing heterologous proteins. If the protein is made in yeast or bacteria, it may be necessary to modify the protein produced therein, for example by phosphorylation or glycosylation of the appropriate sites, in order to obtain the functional protein. Such covalent attachments may be accomplished using known chemical or enzymatic methods.

In another embodiment of the present invention, cells and tissues may be engineered to express an endogenous gene comprising the polynucleotides of the invention under the control of inducible regulatory elements, in which case the regulatory sequences of the endogenous gene may be replaced by homologous recombination. As described herein, gene targeting can be used to replace a gene's existing regulatory region with a regulatory sequence isolated from a different gene or a novel regulatory sequence synthesized by genetic engineering methods. Such regulatory sequences may be comprised of promoters, enhancers, scaffold-attachment regions, negative regulatory elements, transcriptional initiation sites, regulatory protein binding sites or combinations of said sequences. Alternatively, sequences which affect the structure or stability of the RNA or protein produced may be replaced, removed, added, or otherwise modified by targeting. These sequence include polyadenylation signals, mRNA stability elements, splice sites, leader sequences for enhancing or modifying transport or secretion properties of the protein, or other sequences which alter or improve the fumction or stability of protein or RNA molecules.

The targeting event may be a simple insertion of the regulatory sequence, placing the gene under the control of the new regulatory sequence, e.g., inserting a new promoter or enhancer or both upstream of a gene. Alternatively, the targeting event may be a simple deletion of a regulatory element, such as the deletion of a tissue-specific negative regulatory element. Alternatively, the targeting event may replace an existing element; for example, a tissue-specific enhancer can be replaced by an enhancer that has broader or different cell-type specificity than the naturally occurring elements. Here, the naturally occurring sequences are deleted and new sequences are added. In all cases, the identification of the targeting event may be facilitated by the use of one or more selectable marker genes that are contiguous with the targeting DNA, allowing for the selection of cells in which the exogenous DNA has integrated into the host cell genome. The identification of the targeting event may also be facilitated by the use of one or more marker genes exhibiting the property of negative selection, such that the negatively selectable marker is linked to the exogenous DNA, but configured such that the negatively selectable marker flanks the targeting sequence, and such that a correct homologous recombination event with sequences in the host cell genome does not result in the stable integration of the negatively selectable marker. Markers useful for this purpose include the Herpes Simplex Virus thyrnidine kinase (TK) gene or the bacterial xanthine-guanine phosphoribosyl-transferase (gpt) gene.

The gene targeting or gene activation techniques which can be used in accordance with this aspect of the invention are more particularly described in U.S. Pat. No. 5,272,071 to Chappel; U.S. Pat. No. 5,578,461 to Sherwin et al.; International Application No. PCT/US92/09627 (WO93/09222) by Selden et al.; and International Application No. PCT/US90/06436 (WO91/06667) by Skoultchi et al., each of which is incorporated by reference herein in its entirety.

3.6 Polypeptides of the Invention

The isolated polypeptides of the invention include, but are not limited to, a polypeptide comprising: the amino acid sequences set forth as any one of SEQ ID NO: 1-948 or an amino acid sequence encoded by any one of the nucleotide sequences SEQ ID NOs: 1-948 or the corresponding fulll length or mature protein. Polypeptides of the invention also include polypeptides preferably with biological or immunological activity that are encoded by: (a) a polynucleotide having any one of the nucleotide sequences set forth in SEQ ID NOs: 1-948 or (b) polynucleotides encoding any one of the amino acid sequences set forth as SEQ ID NO: 1-948 or (c) polynucleotides that hybridize to the complement of the polynucleotides of either (a) or (b) under stringent hybridization conditions. The invention also provides biologically active or immunologically active variants of any of the amino acid sequences set forth as SEQ ID NO: 1-948 or the corresponding full length or mature protein; and “substantial equivalents” thereof (e.g., with at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, 86%, 87%, 88%, 89%, at least about 90%, 91%, 92%, 93%, 94%, typically at least about 95%, 96%, 97%, more typically at least about 98%, or most typically at least about 99% amino acid identity) that retain biological activity. Polypeptides encoded by allelic variants may have a similar, increased, or decreased activity compared to polypeptides comprising SEQ ID NO: 1-948.

Fragments of the proteins of the present invention which are capable of exhibiting biological activity are also encompassed by the present invention. Fragments of the protein may be in linear form or they may be cyclized using known methods, for example, as described in H. U. Saragovi, et al., Bio/Technology 10, 773-778 (1992) and in R. S. McDowell, et al., J. Amer. Chem. Soc. 114, 9245-9253 (1992), both of which are incorporated herein by reference. Such fragments may be fused to carrier molecules such as immunoglobulins for many purposes, including increasing the valency of protein binding sites.

The present invention also provides both full-length and mature forms (for example, without a signal sequence or precursor sequence) of the disclosed proteins. The protein coding sequence is identified in the sequence listing by translation of the disclosed nucleotide sequences. The mature form of such protein may be obtained by expression of a full-length polynuclcotide in a suitable mammalian cell or other host cell. The sequence of the mature form of the protein is also determinable from the amino acid sequence of the full-length form. Where proteins of the present invention are membrane bound, soluble forms of the proteins are also provided. In such forms, part or all of the regions causing the proteins to be membrane bound are deleted so that the proteins are fully secreted from the cell in which they are expressed.

Protein compositions of the present invention may further comprise an acceptable carrier, such as a hydrophilic, e.g., pharmaceutically acceptable, carrier.

The present invention further provides isolated polypeptides encoded by the nucleic acid fragments of the present invention or by degenerate variants of the nucleic acid fragments of the present invention. By “degenerate variant” is intended nucleotide fragments which differ from a nucleic acid fragment of the present invention (e.g., an ORF) by nucleotide sequence but, due to the degeneracy of the genetic code, encode an identical polypeptide sequence. Preferred nucleic acid fragments of the present invention are the ORFs that encode proteins.

A variety of methodologies kcnown in the art can be utilized to obtain any one of the isolated polypeptides or proteins of the present invention. At the simplest level, the amino acid sequence can be synthesized using commercially available peptide synthesizers. The synthetically-constructed protein sequences, by virtue of sharing primary, secondary or tertiary structural and/or conformational characteristics with proteins may possess biological properties in common therewith, including protein activity. This technique is particularly useful in producing small peptides and fragments of larger polypeptides. Fragments are useful, for example, in generating antibodies against the native polypeptide. Thus, they may be employed as biologically active or immunological substitutes for natural, purified proteins in screening of therapeutic compounds and in immunological processes for the development of antibodies.

The polypeptides and proteins of the present invention can alternatively be purified from cells which have been altered to express the desired polypeptide or protein. As used herein, a cell is said to be altered to express a desired polypeptide or protein when the cell, through genetic manipulation, is made to produce a polypeptide or protein which it normally does not produce or which the cell normally produces at a lower level. One skilled in the art can readily adapt procedures for introducing and expressing either recombinant or synthetic sequences into eukaryotic or prokaryotic cells in order to generate a cell which produces one of the polypeptides or proteins of the present invention.

The invention also relates to methods for producing a polypeptide comprising growing a culture of host cells of the invention in a suitable culture medium, and purifying the protein from the cells or the culture in which the cells are grown. For example, the methods of the invention include a process for producing a polypeptide in which a host cell containing a suitable expression vector that includes a polynucleotide of the invention is cultured under conditions that allow expression of the encoded polypeptide. The polypeptide can be recovered from the culture, conveniently from the culture medium, or from a lysate prepared from the host cells and further purified. Preferred embodiments include those in which the protein produced by such process is a full length or mature form of the protein.

In an alternative method, the polypeptide or protein is purified from bacterial cells which naturally produce the polypeptide or protein. One skilled in the art can readily follow known methods for isolating polypeptides and proteins in order to obtain one of the isolated polypeptides or proteins of the present invention. These include, but are not limited to, immunochromatography, HPLC, size-exclusion chromatography, ion-exchange chromatography, and immuno-affinity chromatography. See, e.g., Scopes, Protein Purification: Principles and Practice , Springer-Verlag (1994); Sambrook, et al., in Molecular Cloning: A Laboratory Manual ; Ausubel et al., Current Protocols in Molecular Biology . Polypeptide fragments that retain biological/immnunological activity include fragments comprising greater than about 100 amino acids, or greater than about 200 amino acids, and fragments that encode specific protein domains.

The purified polypeptides can be used in in vitro binding assays which are well known in the art to identify molecules which bind to the polypeptides. These molecules include but are not limited to, for e.g., small molecules, molecules from combinatorial libraries, antibodies or other proteins. The molecules identified in the binding assay are then tested for antagonist or agonist activity in in vivo tissue culture or animal models that are well known in the art. In brief, the molecules are titrated into a plurality of cell cultures or animals and then tested for either cell/animal death or prolonged survival of the animal/cells.

In addition, the peptides of the invention or molecules capable of binding to the peptides may be complexed with toxins, e.g., ricin or cholera, or with other compounds that are toxic to cells. The toxin-binding molecule complex is then targeted to a tumor or other cell by the specificity of the binding molecule for SEQ ID NO: 1-948.

The protein of the invention may also be expressed as a product of transgenic animals, e.g., as a component of the milk of transgenic cows, goats, pigs, or sheep which are characterized by somatic or germ cells containing a nucleotide sequence encoding the protein.

The proteins provided herein also include proteins characterized by amino acid sequences similar to those of purified proteins but into which modification are naturally provided or deliberately engineered. For example, modifications, in the peptide or DNA sequence, can be made by those skilled in the art using known techniques. Modifications of interest in the protein sequences may include the alteration, substitution, replacement, insertion or deletion of a selected amino acid residue in the coding sequence. For example, one or more of the cysteine residues may be deleted or replaced with another amino acid to alter the conformation of the molecule. Techniques for such alteration, substitution, replacement, insertion or deletion are well known to those skilled in the art (see, e.g., U.S. Pat. No. 4,518,584). Preferably, such alteration, substitution, replacement, insertion or deletion retains the desired activity of the protein. Regions of the protein that are important for the protein function can be determined by various methods known in the art including the alanine-scanning method which involved systematic substitution of single or strings of amino acids with alanine, followed by testing the resulting alanine-containing variant for biological activity. This type of analysis determines the importance of the substituted amino acid(s) in biological activity. Regions of the protein that are important for protein function may be determined by the eMATRIX program.

Other fragments and derivatives of the sequences of proteins which would be expected to retain protein activity in whole or in part and are useful for screening or other immunological methodologies may also be easily made by those skilled in the art given the disclosures herein. Such modifications are encompassed by the present invention.

The protein may also be produced by operably linking the isolated polynucleotide of the invention to suitable control sequences in one or more insect expression vectors, and employing an insect expression system. Materials and methods for baculovirus/insect cell expression systems are commercially available in kit form from, e.g., Invitrogen, San Diego, Calif., U.S.A. (the MaxBa™ kit), and such methods are well known in the art, as described in Summers and Smith, Texas Agricultural Experiment Station Bulletin No. 1555 (1987), incorporated herein by reference. As used herein, an insect cell capable of expressing a polynucleotide of the present invention is “transformed.”

The protein of the invention may be prepared by culturing transformed host cells under culture conditions suitable to express the recombinant protein. The resulting expressed protein may then be purified from such culture (i.e., from culture medium or cell extracts) using known purification processes, such as gel filtration and ion exchange chromatography. The purification of the protein may also include an affinity column containing agents which will bind to the protein; one or more column steps over such affinity resins as concanavalin A-agarose, heparin-toyopearl™ or Cibacrom blue 3GA Sepharose™; one or more steps involving hydrophobic interaction chromatography using such resins as phenyl ether, butyl ether, or propyl ether; or immunoaffinity chromatography.

Alternatively, the protein of the invention may also be expressed in a form which will facilitate purification. For example, it may be expressed as a fulsion protein, such as those of maltose binding protein (MBP), glutathione-S-transferase (GST) or thioredoxin (TRX), or as a His tag. Kits for expression and purification of such fusion proteins are commercially available from New England BioLab (Beverly, Mass.), Pharmacia (Piscataway, N.J.) and Invitrogen, respectively. The protein can also be tagged with an epitope and subsequently purified by using a specific antibody directed to such epitope. One such epitope (“FLAG®”) is commercially available from Kodak (New Haven, Conn.).

Finally, one or more reverse-phase high performance liquid chromatography (RP-BPLC) steps employing hydrophobic RP-HPLC media, e.g. silica gel having pendant methyl or other aliphatic groups, can be employed to further purify the protein. Some or all of the foregoing purification steps, in various combinations, can also be employed to provide a substantially homogeneous isolated recombinant protein. The protein thus purified is substantially free of other mammalian proteins and is defined in accordance with the present invention as an “isolated protein.”

The polypeptides of the invention include analogs (variants). This embraces fragments, as well as peptides in which one or more amino acids has been deleted, inserted, or substituted. Also, analogs of the polypeptides of the invention embrace fusions of the polypeptides or modifications of the polypeptides of the invention, wherein the polypeptide or analog is fused to another moiety or moieties, e.g., targeting moiety or another therapeutic agent. Such analogs may exhibit improved properties such as activity and/or stability. Examples of moieties which may be fused to the polypeptide or an analog include, for example, targeting moieties which provide for the delivery of polypeptide to pancreatic cells, e.g., antibodies to pancreatic cells, antibodies to immune cells such as T-cells, monocytes, dendritic cells, granulocytes, etc., as well as receptor and ligands expressed on pancreatic or immune cells. Other moieties which may be fused to the polypeptide include therapeutic agents which are used for treatment, for example, immunosuppressive drugs such as cyclosporin, SK506, azathioprine, CD3 antibodies and steroids. Also, polypeptides may be fused to immune modulators, and other cytokines such as alpha or beta interferon.

3.6.1 Determining Polypeptide and Polynucleotide Identity and Similarity

Preferred identity and/or similarity are designed to give the largest match between the sequences tested. Methods to determine identity and similarity are codified in computer programs including, but are not limited to, the GCG program package, including GAP (Devereux, J., et al., Nucleic Acids Research 12(1):387 (1984); Genetics Computer Group, University of Wisconsin, Madison, Wis.), BLASTP, BLASTN, BLASTX, FASTA (Altschul, S. F. et al., J. Molec. Biol. 215:403-410 (1990), PSI-BLAST (Altschul S. F. et al., Nucleic Acids Res. vol. 25, pp. 3389-3402, herein incorporated by reference), eMatrix software (Wu et al., J. Comp. Biol., Vol. 6, pp. 219-235 (1999), herein incorporated by reference), eMotif software (Nevill-Manning et al, ISMB-97, Vol. 4, pp. 202-209, herein incorporated by reference), pFam software (Sornhammer et al., Nucleic Acids Res., Vol. 26(1), pp. 320-322 (1998), herein incorporated by reference) and the Kyte-Doolittle hydrophobocity prediction algorithm (J. Mol Biol, 157, pp. 105-31 (1982), incorporated herein by reference). The BLAST programs are publicly available from the National Center for Biotechnology Information (NCBI) and other sources (BLAST Manual, Altschul, S., et al. NCB NLM NIH Bethesda, Md. 20894; Altschul, S., et al., J. Mol. Biol. 215:403-410 (1990).

3.7 Chimeric and Fusion Proteins

The invention also provides chimeric or fusion proteins. As used herein, a “chimeric protein” or “fusion protein” comprises a polypeptide of the invention operatively linked to another polypeptide. Within a fusion protein the polypeptide according to the invention can correspond to all or a portion of a protein according to the invention. In one embodiment, a fusion protein comprises at least one biologically active portion of a protein according to the invention. In another embodiment, a fusion protein comprises at least two biologically active portions of a protein according to the invention. Within the fusion protein, the term “operatively linked” is intended to indicate that the polypeptide according to the invention and the other polypeptide are fused in-frame to each other. The polypeptide can be fused to the N-terminus or C-terminus, or to the middle.

For example, in one embodiment a fusion protein comprises a polypeptide according to the invention operably linked to the extracellular domain of a second protein.

In another embodiment, the fusion protein is a GST-fusion protein in which the polypeptide sequences of the invention are fused to the C-terminus of the GST (i.e., glutathione S-transferase) sequences.

In another embodiment, the fusion protein is an immunoglobulin fusion protein in which the polypeptide sequences according to the invention comprise one or more domains fused to sequences derived from a member of the immunoglobulin protein family. The immnunoglobulin fusion proteins of the invention can be incorporated into pharmaceutical compositions and administered to a subject to inhibit an interaction between a ligand and a protein of the invention on the surface of a cell, to thereby suppress signal transduction in vivo. The immunoglobulin fusion proteins can be used to affect the bioavailability of a cognate ligand. Inhibition of the ligand/protein interaction may be useful therapeutically for both the treatment of proliferative and differentiative disorders, e.g., cancer as well as modulating (e.g., promoting or inhibiting) cell survival. Moreover, the immunoglobulin fusion proteins of the invention can be used as immunogens to produce antibodies in a subject, to purify ligands, and in screening assays to identify molecules that inhibit the interaction of a polypeptide of the invention with a ligand.

A chimeric or fiusion protein of the invention can be produced by standard recombinant DNA techniques. For example, DNA fragments coding for the different polypeptide sequences are ligated together in-frame in accordance with conventional techniques, e.g., by employing blunt-ended or stagger-ended termini for ligation, restriction enzyme digestion to provide for appropriate termini, filling-in of cohesive ends as appropriate, alkaline phosphatase treatment to avoid undesirable joining, and enzymatic ligation. In another embodiment, the fusion gene can be synthesized by conventional techniques including automated DNA synthesizers. Alternatively, PCR amplification of gene fragments can be carried out using anchor primers that give rise to complementary overhangs between two consecutive gene fragments that can subsequently be annealed and reamplified to generate a chimeric gene sequence (see, for example, Ausubel et al. (eds.) C URRENT P ROTOCOLS IN M OLECULAR B IOLOGY , John Wiley & Sons, 1992). Moreover, many expression vectors are commercially available that already encode a fusion moiety (e.g., a GST polypeptide). A nucleic acid encoding a polypeptide of the invention can be cloned into such an expression vector such that the fusion moiety is linked in-frame to the protein of the invention.

3.8 Gene Therapy

Mutations in the polynucleotides of the invention gene may result in loss of normal function of the encoded protein. The invention thus provides gene therapy to restore normal activity of the polypeptides of the invention; or to treat disease states involving polypeptides of the invention. Delivery of a functional gene encoding polypeptides of the invention to appropriate cells is effected ex vivo, in situ, or in vivo by use of vectors, and more particularly viral vectors (e.g., adenovirus, adeno-associated virus, or a retrovirus), or ex vivo by use of physical DNA transfer methods (e.g., liposomes or chemical treatments). See, for example, Anderson, Nature, supplement to vol. 392, no. 6679, pp.25-20 (1998). For additional reviews of gene therapy technology see Friedmann, Science, 244: 1275-1281 (1989); Verma, Scientific American: 68-84 (1990); and Miller, Nature, 357: 455460(1992). Introduction of any one of the nucleotides of the present invention or a gene encoding the polypeptides of the present invention can also be accomplished with extrachromosomal substrates (transient expression) or artificial chromosomes (stable expression). Cells may also be cultured ex vivo in the presence of proteins of the present invention in order to proliferate or to produce a desired effect on or activity in such cells. Treated cells can then be introduced in vivo for therapeutic purposes. Alternatively, it is contemplated that in other human disease states, preventing the expression of or inhibiting the activity of polypeptides of the invention will be useful in treating the disease states. It is contemplated that antisense therapy or gene therapy could be applied to negatively regulate the expression of polypeptides of the invention.

Other methods inhibiting expression of a protein include the introduction of antisense molecules to the nucleic acids of the present invention, their complements, or their translated RNA sequences, by methods known in the art. Further, the polypeptides of the present invention can be inhibited by using targeted deletion methods, or the insertion of a negative regulatory element such as a silencer, which is tissue specific.

The present invention still further provides cells genetically engineered in vivo to express the polynucleotides of the invention, wherein such polynucleotides are in operative association with a regulatory sequence heterologous to the host cell which drives expression of the polynucleotides in the cell. These methods can be used to increase or decrease the expression of the polynucleotides of the present invention.

Knowledge of DNA sequences provided by the invention allows for modification of cells to permit, increase, or decrease, expression of endogenous polypeptide. Cells can be modified (e.g., by homologous recombination) to provide increased polypeptide expression by replacing, in whole or in part, the naturally occurring promoter with all or part of a heterologous promoter so that the cells express the protein at higher levels. The heterologous promoter is inserted in such a manner that it is operatively linked to the desired protein encoding sequences. See, for example, PCT International Publication No. WO 94/12650, PCT International Publication No. WO 92/20808, and PCT International Publication No. WO 91/09955. It is also contemplated that, in addition to heterologous promoter DNA, amplifiable marker DNA (e.g., ada, dhfr, and the multifunctional CAD gene which encodes carbamyl phosphate synthase, aspartate transcarbamylase, and dihydroorotase) and/or intron DNA may be inserted along with the heterologous promoter DNA. If linked to the desired protein coding sequence, amplification of the marker DNA by standard selection methods results in co-amplification of the desired protein coding sequences in the cells.

In another embodiment of the present invention, cells and tissues may be engineered to express an endogenous gene comprising the polynucleotides of the invention under the control of inducible regulatory elements, in which case the regulatory sequences of the endogenous gene may be replaced by homologous recombination. As described herein, gene targeting can be used to replace a gene's existing regulatory region with a regulatory sequence isolated from a different gene or a novel regulatory sequence synthesized by genetic engineering methods. Such regulatory sequences may be comprised of promoters, enhancers, scaffold-attachment regions, negative regulatory elements, transcriptional initiation sites, regulatory protein binding sites or combinations of said sequences. Alternatively, sequences which affect the structure or stability of the RNA or protein produced may be replaced, removed, added, or otherwise modified by targeting. These sequences include polyadenylation signals, mRNA stability elements, splice sites, leader sequences for enhancing or modifying transport or secretion properties of the protein, or other sequences which alter or improve the function or stability of protein or RNA molecules.

The targeting event may be a simple insertion of the regulatory sequence, placing the gene under the control of the new regulatory sequence, e.g., inserting a new promoter or enhancer or both upstream of a gene. Alternatively, the targeting event may be a simple deletion of a regulatory element, such as the deletion of a tissue-specific negative regulatory element. Alternatively, the targeting event may replace an existing element; for example, a tissue-specific enhancer can be replaced by an enhancer that has broader or different cell-type specificity than the naturally occurring elements. Here, the naturally occurring sequences are deleted and new sequences are added. In all cases, the identification of the targeting event may be facilitated by the use of one or more selectable marker genes that are contiguous with the targeting DNA, allowing for the selection of cells in which the exogenous DNA has integrated into the cell genome. The identification of the targeting event may also be facilitated by the use of one or more marker gencs exhibiting the property of negative selection, such that the negatively selectable marker is linked to the exogenous DNA, but configured such that the negatively selectable marker flanks the targeting sequence, and such that a correct homologous recombination event with sequences in the host cell genome does not result in the stable integration of the negatively selectable marker. Markers useful for this purpose include the Herpes Simplex Virus thyrnidine kinase (TK) gene or the bacterial xanthine-guanine phosphoribosyl-transferase (gpt) gene.

The gene targeting or gene activation techniques which can be used in accordance with this aspect of the invention are more particularly described in U.S. Pat. No. 5,272,071 to Chappel; U.S. Pat. No. 5,578,461 to Sherwin et al.; International Application No. PCT/US92/09627 (WO93/09222) by Selden et al.; and International Application No. PCT/US90/06436 (WO91/06667) by Skoultchi et al., each of which is incorporated by reference herein in its entirety.

3.9 Transgenic Animals

In preferred methods to determine biological functions of the polypeptides of the invention in vivo, one or more genes provided by the invention are either over expressed or inactivated in the germ line of animals using homologous recombination [Capecchi, Science 244:1288-1292 (1989)]. Animals in which the gene is over expressed, under the regulatory control of exogenous or endogenous promoter elements, are known as transgenic animals. Animals in which an endogenous gene has been inactivated by homologous recombination are referred to as “knockout” animals. Knockout animals, preferably non-human mammals, can be prepared as described in U.S. Pat. No. 5,557,032, incorporated herein by reference. Transgenic animals are useful to determine the roles polypeptides of the invention play in biological processes, and preferably in disease states. Transgenic animals are useful as model systems to identify compounds that modulate lipid metabolism. Transgenic animals, preferably non-human mammals, are produced using methods as described in U.S. Pat. No. 5,489,743 and PCT Publication No. WO94/28122, incorporated herein by reference.

Transgenic animals can be prepared wherein all or part of a promoter of the polynucleotides of the invention is either activated or inactivated to alter the level of expression of the polypeptides of the invention. Inactivation can be carried out using homologous recombination methods described above. Activation can be achieved by supplementing or even replacing the homologous promoter to provide for increased protein expression. The homologous promoter can be supplemented by insertion of one or more heterologous enhancer elements known to confer promoter activation in a particular tissue.

The polynucleotides of the present invention also make possible the development, through, e.g., homologous recombination or knock out strategies, of animals that fail to express polypeptides of the invention or that express a variant polypeptide. Such animals are useful as models for studying the in vivo activities of polypeptide as well as for studying modulators of the polypeptides of the invention.

In preferred methods to determine biological functions of the polypeptides of the invention in vivo, one or more genes provided by the invention are either over expressed or inactivated in the germ line of animals using homologous recombination [Capecchi, Science 244:1288-1292 (1989)]. Animals in which the gene is over expressed, under the regulatory control of exogenous or endogenous promoter elements, are known as transgenic animals. Animals in which an endogenous gene has been inactivated by homologous recombination are referred to as “knockout” animals. Knockout animals, preferably non-human mammals, can be prepared as described in U.S. Pat. No. 5,557,032, incorporated herein by reference. Transgenic animals are useful to determine the roles polypeptides of the invention play in biological processes, and preferably in disease states. Transgenic animals are useful as model systems to identify compounds that modulate lipid metabolism. Transgenic animals, preferably non-human mammals, are produced using methods as described in U.S. Pat. No. 5,489,743 and PCT Publication No. WO94/28122, incorporated herein by reference.

Transgenic animals can be prepared wherein all or part of the polynucleotides of the invention promoter is either activated or inactivated to alter the level of expression of the polypeptides of the invention. Inactivation can be carried out using homologous recombination methods described above. Activation can be achieved by supplementing or even replacing the homologous promoter to provide for increased protein expression. The homologous promoter can be supplemented by insertion of one or more heterologous enhancer elements known to confer promoter activation in a particular tissue.

3.10 Uses and Biological Activity

The polynucleotides and proteins of the present invention are expected to exhibit one or more of the uses or biological activities (including those associated with assays cited herein) identified herein. Uses or activities described for proteins of the present invention may be provided by administration or use of such proteins or of polynuclcotides encoding such proteins (such as, for example, in gene therapies or vectors suitable for introduction of DNA). The mechanism underlying the particular condition or pathology will dictate whether the polypeptides of the invention, the polynucleotides of the invention or modulators (activators or inhibitors) thereof would be beneficial to the subject in need of treatment. Thus, “therapeutic compositions of the invention” include compositions comprising isolated polynucleotides (including recombinant DNA molecules, cloned genes and degenerate variants thereof) or polypeptides of the invention (including full length protein, mature protein and truncations or domains thereof), or compounds and other substances that modulate the overall activity of the target gene products, either at the level of target gene/protein expression or target protein activity. Such modulators include polypeptides, analogs, (variants), including fragments and fusion proteins, antibodies and other binding proteins; chemical compounds that directly or indirectly activate or inhibit the polypeptides of the invention (identified, e.g., via drug screening assays as described herein); antisense polynucleotides and polynucleotides suitable for triple helix formation; and in particular antibodies or other binding partners that specifically recogrize one or more epitopes of the polypeptides of the invention.

The polypeptides of the present invention may likewise be involved in cellular activation or in one of the other physiological pathways described herein.

3.10.1 Research Uses and Utilities

The polynucleotides provided by the present invention can be used by the research community for various purposes. The polynucleotides can be used to express recombinant protein for analysis, characterization or therapeutic use; as markers for tissues in which the corresponding protein is preferentially expressed (either constitutively or at a particular stage of tissue differentiation or development or in disease states); as molecular weight markers on gels; as chromosome markers or tags (when labeled) to identify chromosomes or to map related gene positions; to compare with endogenous DNA sequences in patients to identify potential genetic disorders; as probes to hybridize and thus discover novel, related DNA sequences; as a source of information to derive PCR primers for genetic fingerprinting; as a probe to “subtract-out” known sequences in the process of discovering other novel polynucleotides; for selecting and making oligomers for attachment to a “gene chip” or other support, including for examination of expression patterns; to raise anti-protein antibodies using DNA immunization techniques; and as an antigen to raise anti-DNA antibodies or elicit another immune response. Where the polynucleotide encodes a protein which binds or potentially binds to another protein (such as, for example, in a receptor-ligand interaction), the polynucleotide can also be used in interaction trap assays (such as, for example, that described in Gyuris et al., Cell 75:791-803 (1993)) to identify polynucleotides encoding the other protein with which binding occurs or to identify inhibitors of the binding interaction.

The polypeptides provided by the present invention can similarly be used in assays to determine biological activity, including in a panel of multiple proteins for high-throughput screening; to raise antibodies or to elicit another immune response; as a reagent (including the labeled reagent) in assays designed to quantitatively determine levels of the protein (or its receptor) in biological fluids; as markers for tissues in which the corresponding polypeptide is preferentially expressed (either constitutively or at a particular stage of tissue differentiation or development or in a disease state); and, of course, to isolate correlative receptors or ligands. Proteins involved in these binding interactions can also be used to screen for peptide or small molecule inhibitors or agonists of the binding interaction.

Any or all of these research utilities are capable of being developed into reagent grade or kit format for corrjercialization as research products.

Methods for performing the uses listed above are well known to those skilled in the art. References disclosing such methods include without limitation “Molecular Cloning: A Laboratory Manual”, 2d ed., Cold Spring Harbor Laboratory Press, in Sambrook, J., E. F. Fritsch and T. Maniatis eds., 1989, and “Methods in Enzymology: Guide to Molecular Cloning Techniques”, Academic Press, Berger, S. L. and A. R Kimmel eds., 1987.

3.10.2 Nutritional Uses

Polynucleotides and polypeptides of the present invention can also be used as nutritional sources or supplements. Such uses include without limitation use as a protein or amino acid supplement, use as a carbon source, use as a nitrogen source and use as a source of carbohydrate. In such cases the polypeptidc or polynucleotide of the invention can be added to the feed of a particular organism or can be administered as a separate solid or liquid preparation, such as in the form of powder, pills, solutions, suspensions or capsules. In the case of microorganisms, the polypeptide or polynucleotide of the invention can be added to the medium in or on which the microorganism is cultured.

3.10.3 Cytokine and Cell Proliferation/Differentiation Activity

A polypeptide of the present invention may exhibit activity relating to cytokine, cell proliferation (either inducing or inhibiting) or cell differentiation (either inducing or inhibiting) activity or may induce production of other cytokines in certain cell populations. A polynucleotide of the invention can encode a polypeptide exhibiting such attributes. Many protein factors discovered to date, including all known cytokines, have exhibited activity in one or more factor-dependent cell proliferation assays, and hence the assays serve as a convenient confirmation of cytokine activity. The activity of therapeutic compositions of the present invention is evidenced by any one of a number of routine factor dependent cell proliferation assays for cell lines including, without limitation, 32D, DA2, DA1G, T10, B9, B9/11, BaF3, MC9/G, M+(preB M+), 2E8, RB5, DA1, 123, T1165, HT2, CTLL2, TF-1, Mo7e, CMK, HUVEC, and Caco. Therapeutic compositions of the invention can be used in the following:

Assays for T-cell or thymocyte proliferation include without limitation those described in: Current Protocols in Immunology, Ed by J. E. Coligan, A. M. Kruisbeek, D. H. Margulies, E. M. Shevach, W. Strober, Pub. Greene Publishing Associates and Wiley-Interscience (Chapter 3, In Vitro assays for Mouse Lymphocyte Function 3.1-3.19; Chapter 7, Immunologic studies in Humans); Takai et al., J. Immunol. 137:3494-3500, 1986; Bertagnolli et al., J. Immunol. 145:1706-1712, 1990; Bertagnolli et al., Cellular Imrnunology 133:327-341, 1991; Bertagnolli, et al., I. Immunol. 149:3778-3783, 1992; Bowman et al., I. Immunol. 152:1756-1761, 1994.

Assays for cytokine production and/or proliferation of spleen cells, lymph node cells or thymocytes include, without limitation, those described in: Polyclonal T cell stimulation, Kruisbeek, A. M. and Shevach, E. M. In Current Protocols in Immunology. J. E. e.a. Coligan eds. Vol 1 pp. 3.12.1-3.12.14, John Wiley and Sons, Toronto. 1994; and Measurement of mouse and human interleukin-y, Schreiber, R. D. In Current Protocols in Immunology. J. E. e.a. Coligan eds. Vol 1 pp. 6.8.1-6.8.8, John Wiley and Sons, Toronto. 1994.

Assays for proliferation and differentiation of hematopoietic and lymphopoietic cells include, without limitation, those described in: Measurement of Human and Murine Interleukin 2 and Interleukin 4, Bottomly, K., Davis, L. S. and Lipsky, P. E. In Current Protocols in Immunology. J. E. e.a. Coligan eds. Vol 1 pp. 6.3.1-6.3.12, John Wiley and Sons, Toronto. 1991; devries et al., J. Exp. Med. 173:1205-1211, 1991; Moreau et al., Nature 336:690-692, 1988; Greenberger et al., Proc. Natl. Acad. Sci. U.S.A. 80:2931-2938, 1983; Measurement of mouse and human interleukin 6—Nordan, R. In Current Protocols in Immunology. J. E. Coligan eds. Vol 1 pp. 6.6.1-6.6.5, John Wilcy and Sons, Toronto. 1991; Smith et al., Proc. Natl. Aced. Sci. U.S.A. 83:1857-1861, 1986; Measurement of human Interleukin 11—Bennett, F., Giannotti, J., Clark, S. C. and Turner, K. J. In Current Protocols in Immunology. J. E. Coligan eds. Vol 1 pp. 6.15.1 John Wiley and Sons, Toronto. 1991; Measurement of mouse and human Interlcukin 9—Ciarletta, A., Giannotti, J., Clark, S. C. and Turner, K. J. In Current Protocols in Imnnunology. J. E. Coligan eds. Vol I pp. 6.13.1, John Wiley and Sons, Toronto. 1991.

Assays for T-cell clone responses to antigens (which will identify, among others, proteins that affect APC-T cell interactions as well as direct T-cell effects by measuring proliferation and cytokine production) include, without limitation, those described in: A Current Protocols in Immunology, Ed by J. E. Coligan, A. M. Kruisbeek, D. H. Margulies, E. M. Shevach, W Strober, Pub. Greene Publishing Associates and Wiley-Interscience (Chapter 3, In Vitro assays for Mouse Lymphocyte Function; Chapter 6, Cytokines and their cellular receptors; Chapter 7, Immunologic studies in Humans); Weinberger et al., Proc. Natl. Acad. Sci. USA 77:6091-6095, 1980; Weinberger et al., Eur. J. Immun. 11:405-411, 1981; Takai et al., J. Immunol. 137:3494-3500, 1986; Takai et al., J. Immunol. 140:508-512, 1988.

3.10.4 Stem Cell Growth Factor Activity

A polypeptide of the present invention may exhibit stem cell growth factor activity and be involved in the proliferation, differentiation and survival of pluripotent and totipotent stem cells including primordial germ cells, embryonic stem cells, hematopoietic stem cells and/or germ line stem cells. Administration of the polypeptide of the invention to stem cells in vivo or ex vivo is expected to maintain and expand cell populations in a totipotential or pluripotential state which would be useful for re-engineering damaged or diseased tissues, transplantation, manufacture of bio-pharmaceuticals and the development of bio-sensors. The ability to produce large quantities of human cells has important working applications for the production of human proteins which currently must be obtained from non-human sources or donors, implantation of cells to treat diseases such as Parkinson's, Alzheimer's and other neurodegenerative diseases; tissues for grafting such as bone marrow, skin, cartilage, tendons, bone, muscle (including cardiac muscle), blood vessels, cornea, neural cells, gastrointestinal cells and others; and organs for transplantation such as kidney, liver, pancreas (including islet cells), heart and lung.

It is contemplated that multiple different exogenous growth factors and/or cytokines may be administered in combination with the polypeptide of the invention to achieve the desired effect, including any of the growth factors listed herein, other stem cell maintenance factors, and specifically including stem cell factor (SCF), leukemia inhibitory factor (LIF), Flt-3 ligand (Flt-3L), any of the interleukins, recombinant soluble IL-6 receptor fused to IL-6, macrophagc inflammatory protein 1-alpha (MIP-1-alpha), G-CSF, GM-CSF, thrombopoietin (TPO), platelet factor 4 (PF-4), platelet-derived growth factor (PDGF), neural growth factors and basic fibroblast growth factor (bFGF).

Since totipotent stem cells can give rise to virtually any mature cell type, expansion of these cells in culture will facilitate the production of large quantities of mature cells. Techniques for culturing stem cells are known in the art and administration of polypeptides of the invention, optionally with other growth factors and/or cytokines, is expected to enhance the survival and proliferation of the stem cell populations. This can be accomplished by direct administration of the polypeptide of the invention to the culture medium. Alternatively, stroma cells transfected with a polynucleotide that encodes for the polypeptide of the invention can be used as a feeder layer for the stem cell populations in culture or in vivo. Stromal support cells for feeder layers may include embryonic bone marrow fibroblasts, bone marrow stromal cells, fetal liver cells, or cultured embryonic fibroblasts (see U.S. Pat. No. 5,690,926).

Stem cells themselves can be transfected with a polynucleotide of the invention to induce autocrine expression of the polypeptide of the invention. This will allow for generation of undifferentiated totipotential/pluripotential stem cell lines that are useful as is or that can then be differentiated into the desired mature cell types. These stable cell lines can also serve as a source of undifferentiated totipotential/pluripotential mRNA to create cDNA libraries and templates for polymerase chain reaction experiments. These studies would allow for the isolation and identification of differentially expressed genes in stem cell populations that regulate stem cell proliferation and/or maintenance.

Expansion and maintenance of totipotent stem cell populations will be useful in the treatment of many pathological conditions. For example, polypeptides of the present invention may be used to manipulate stem cells in culture to give rise to neuroepithelial cells that can be used to augment or replace cells damaged by illness, autoimmune disease, accidental damage or genetic disorders. The polypeptide of the invention may be useful for inducing the proliferation of neural cells and for the regeneration of nerve and brain tissue, i.e. for the treatment of central and peripheral nervous system diseases and neuropathies, as well as mechanical and traumatic disorders which involve degeneration, death or trauma to neural cells or nerve tissue. In addition, the expanded stem cell populations can also be genetically altered for gene therapy purposes and to decrease host rejection of replacement tissues after grafting or implantation.

Expression of the polypeptide of the invention and its effect on stem cells can also be manipulated to achieve controlled differentiation of the stem cells into more differentiated cell types. A broadly applicable method of obtaining pure populations of a specific differentiated cell type from undifferentiated stem cell populations involves the use of a cell-type specific promoter driving a selectable marker. The selectable marker allows only cells of the desired type to survive. For example, stem cells can be induced to differentiate into cardiomyocytes (Wobus et al., Differentiation, 48: 173-182, (1991); Klug et al., J. Clin. Invest., 98(1): 216-224, (1998)) or skeletal muscle cells (Browder, L. W. In: Principles of Tissue Engineering eds . Lanza et al., Acadernic Press (1997)). Alternatively, directed differentiation of stem cells can be accomplished by culturing the stem cells in the presence of a differentiation factor such as retinoic acid and an antagonist of the polypeptide of the invention which would inhibit the effects of endogenous stem cell factor activity and allow differentiation to proceed.

In vitro cultures of stem cells can be used to determine if the polypeptide of the invention exhibits stem cell growth factor activity. Stem cells are isolated from any one of various cell sources (including hematopoietic stem cells and embryonic stem cells) and cultured on a feeder layer, as described by Thompson et al. Proc. Natl. Acad. Sci, U.S.A., 92: 7844-7848 (1995), in the presence of the polypeptide of the invention alone or in combination with other growth factors or cytokines. The ability of the polypeptide of the invention to induce stem cells proliferation is determined by colony formation on semi-solid support e.g. as described by Bernstein et al., Blood, 77: 2316-2321 (1991).

3.10.5 Hematopoiesis Regulating Activity

A polypeptide of the present invention may be involved in regulation of hematopoiesis and, consequently, in the treatment of myeloid or lymphoid cell disorders. Even marginal biological activity in support of colony forming cells or of factor-dependent cell lines indicates involvement in regulating hematopoiesis, e.g. in supporting the growth and proliferation of erythwid progenitor cells alone or in combination with other cytokines, thereby indicating utility, for example, in treating various anemias or for use in conjunction with irradiation/chemotherapy to stimulate the production of erythroid precursors and/or erythroid cells; in supporting the growth and proliferation of myeloid cells such as granulocytes and monocytes/macrophages (i.e., traditional CSF activity) useful, for example, in conjunction with chemotherapy to prevent or treat consequent myelo-suppression; in supporting the growth and proliferation of megakaryocytes and consequently of platelets thereby allowing prevention or treatment of various platelet disorders such as thrombocytopenia, and generally for use in place of or complimentary to platelet transfusions; and/or in supporting the growth and proliferation of hematopoietic stem cells which are capable of maturing to any and all of the above-mentioned hematopoietic cells and therefore find therapeutic utility in various stem cell disorders (such as those usually treated with transplantation, including, without limitation, aplastic anemia and paroxysmal nocturnal hemoglobinuria), as well as in repopulating the stem cell compartment post irradiation/chemotherapy, either in-vivo or ex-vivo (i.e., in conjunction with bone marrow transplantation or with peripheral progenitor cell transplantation (homologous or heterologous)) as normal cells or genetically manipulated for gene therapy.

Therapeutic compositions of the invention can be used in the following:

Suitable assays for proliferation and differentiation of various hematopoietic lines are cited above.

Assays for embryonic stem cell differentiation (which will identify, among others, proteins that influence embryonic differentiation hematopoiesis) include, without limitation, those described in: Johansson et al. Cellular Biology 15:141-151, 1995; Keller et al., Molecular and Cellular Biology 13:473-486, 1993; McClanahan et al., Blood 81:2903-2915, 1993.

Assays for stem cell survival and differentiation (which will identify, among others, proteins that regulate lympho-hematopoiesis) include, without limitation, those described in: Methylcellulose colony formring assays, Freshney, M. G. In Culture of Hematopoietic Cells. R. L. Freshney, et al. eds. Vol pp. 265-268, Wiley-Liss, Inc., New York, N.Y. 1994; Hirayama et al., Proc. Natl. Acad. Sci. USA 89:5907-5911, 1992; Primitive hematopoietic colony forming cells with high proliferative potential, McNiece, I. K. and Briddell, R. A. In Culture of Hematopoietic Cells. R. I. Freshney, et al. eds. Vol pp. 23-39, Wiley-Liss, Inc., New York, N.Y. 1994; Neben et al., Experimental Hematology 22:353-359, 1994; Cobblestone area forming cell assay, Ploemacher, R. E. In Culture of Hematopoietic Cells. R. I. Freshney, et al. eds. Vol pp. 1-21, Wiley-Liss, Inc., New York, N.Y. 1994; Long term bone marrow cultures in the presence of stromal cells, Spooncer, E., Dexter, M. and Allen, T. In Culture of Hematopoietic Cells. R. I. Freshney, et al. eds. Vol pp. 163-179, Wiley-Liss, Inc., New York, N.Y. 1994; Long term culture initiating cell assay, Sutherland, H. J. In Culture of Hematopoietic Cells. R I. Freshney, et al. eds. Vol pp. 139-162, Wiley-Liss, Inc., New York, N.Y. 1994.

3.10.6 Tissue Growth Activity

A polypeptide of the present invention also may be involved in bone, cartilage, tendon, ligament and/or nerve tissue growth or regeneration, as well as in wound healing and tissue repair and replacement, and in healing of bums, incisions and ulcers.

A polypeptide of the present invention which induces cartilage and/or bone growth in circumstances where bone is not normally formed, has application in the healing of bone fractures and cartilage damage or defects in humans and other animals. Compositions of a polypeptide, antibody, binding partner, or other modulator of the invention may have prophylactic use in closed as well as open fracture reduction and also in the improved fixation of artificial joints. De novo bone formation induced by an osteogenic agent contributes to the repair of congenital, trauma induced, or oncologic resection induced craniofacial defects, and also is useful in cosmetic plastic surgery.

A polypeptide of this invention may also be involved in attracting bone-forming cells, stimulating growth of bone-formning cells, or inducing differentiation of progenitors of bone-forming cells. Treatment of osteoporosis, osteoarthritis, bone degenerative disorders, or periodontal disease, such as through stimulation of bone and/or cartilage repair or by blocking inflammation or processes of tissue destruction (collagenase activity, osteoclast activity, etc.) mediated by inflammatory processes may also be possible using the composition of the invention.

Another category of tissue regeneration activity that may involve the polypeptide of the present invention is tendon/ligament formation. Induction of tendon/ligament-like tissue or other tissue formation in circumstances where such tissue is not normally formed, has application in the healing of tendon or ligament tears, deformities and other tendon or ligament defects in humans and other animals. Such a preparation employing a tendon/ligament-like tissue inducing protein may have prophylactic use in preventing damage to tendon or ligament tissue, as well as use in the improved fixation of tendon or ligament to bone or other tissues, and in repairing defects to tendon or ligament tissue. De novo tendon/ligament-like tissue formation induced by a composition of the present invention contributes to the repair of congenital, trauma induced, or other tendon or ligament defects of other origin, and is also useful in cosmetic plastic surgery for attachment or repair of tendons or ligaments. The compositions of the present invention may provide environment to attract tendon- or ligament-forrning cells, stimulate growth of tendon- or ligament-forming cells, induce differentiation of progenitors of tendon- or ligament-forming cells, or induce growth of tendon/liganent cells or progenitors ex vivo for return in vivo to effect tissue repair. The compositions of the invention may also be useful in the treatment of tendinitis, carpal tunnel syndrome and other tendon or ligament defects. The compositions may also include an appropriate matrix and/or sequestering agent as a carrier as is well known in the art.

The compositions of the present invention may also be useful for proliferation of neural cells and for regeneration of nerve and brain tissue, i.e. for the treatment of central and peripheral nervous system diseases and neuropathies, as well as mechanical and traumatic disorders, which involve degeneration, death or trauma to neural cells or nerve tissue. More specifically, a composition may be used in the treatment of diseases of the peripheral nervous system, such as peripheral nerve injuries, peripheral neuropathy and localized neuropathies, and central nervous system diseases, such as Alzheimer's, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis, and Shy-Drager syndrome. Further conditions which may be treated in accordance with the present invention include mechanical and traumatic disorders, such as spinal cord disorders, head trauma and cerebrovascular diseases such as stroke. Peripheral neuropathies resulting from chemotherapy or other medical therapies may also be treatable using a composition of the invention.

Compositions of the invention may also be useful to promote better or faster closure of non-healing wounds, including without limitation pressure ulcers, ulcers associated with vascular insufficiency, surgical and traumatic wounds, and the like.

Compositions of the present invention may also be involved in the generation or regeneration of other tissues, such as organs (including, for example, pancreas, liver, intestine, kidney, skin, endothelium), muscle (smooth, skeletal or cardiac) and vascular (including vascular endothelium) tissue, or for promoting the growth of cells comprising such tissues. Part of the desired effects may be by inhibition or modulation of fibrotic scarring may allow normal tissue to regenerate. A polypeptide of the present invention may also exhibit angiogenic activity.

A composition of the present invention may also be useful for gut protection or regeneration and treatment of lung or liver fibrosis, reperfusion injury in various tissues, and conditions resulting from systemic cytokine damage.

A composition of the present invention may also be useful for promoting or inhibiting differentiation of tissues described above from precursor tissues or cells; or for inhibiting the growth of tissues described above.

Therapeutic compositions of the invention can be used in the following:

Assays for tissue generation activity include, without limitation, those described in: International Patent Publication No. WO95/16035 (bone, cartilage, tendon); International Patent Publication No. WO95/05846 (nerve, neuronal); International Patent Publication No. WO91/07491 (skin, endothelium).

Assays for wound healing activity include, without limitation, those described in: Winter, Epidermal Wound Healing, pps. 71-112 (Maibach, H. L. and Rovee, D. T., eds.), Year Book Medical Publishers, Inc., Chicago, as modified by Eaglstein and Mertz, J. Invest. Dermatol 71:382-84 (1978).

3.10.7 Immune Stimulating or Suppressing Activity

A polypeptide of the present invention may also exhibit immune stimulating or immune suppressing activity, including without limitation the activities for which assays are described herein. A polynucleotide of the invention can encode a polypeptide exhibiting such activities. A protein may be useful in the treatment of various immune deficiencies and disorders (including severe combined immunodeficiency (SCID)), e.g., in regulating (up or down) growth and proliferation of T and/or B lymphocytes, as well as effecting the cytolytic activity of NK cells and other cell populations. These immune deficiencies may be genetic or be caused by viral (e.g., HI) as well as bacterial or fungal infections, or may result from autoimmune disorders. Morc specifically, infectious diseases causes by viral, bacterial, ftngal or other infection may be treatable using a protein of the present invention, including infections by HIV, hepatitis viruses, herpes viruses, mycobacteria, Leishmania spp., malaria spp. and various fungal infections such as candidiasis. Of course, in this regard, proteins of the present invention may also be useful where a boost to the immune system generally may be desirable, i.e., in the treatment of cancer.

Autoimmune disorders which may be treated using a protein of the present invention include, for example, connective tissue disease, multiple sclerosis, systemic lupus erythematosus, rheumatoid arthritis, autoimmune pulmonary inflammation, Guillain-Barre syndrome, autoimmune thyroiditis, insulin dependent diabetes mellitis, myasthenia gravis, graft-versus-host disease and autoimmune inflammatory eye disease. Such a protein (or antagonists thereof, including antibodies) of the present invention may also to be useful in the treatment of allergic reactions and conditions (e.g., anaphylaxis, serum sickness, drug reactions, food allergies, insect venom allergies, mastocytosis, allergic rhinitis, hypersensitivity pneumonitis, urticaria, angioedema, eczema, atopic dermatitis, allergic contact dermatitis, erythema multiforme, Stevens-Johnson syndrome, allergic conjunctivitis, atopic keratoconjunctivitis, venereal keratoconjunctivitis, giant papillary conjunctivitis and contact allergies), such as asthma (particularly allergic asthma) or other respiratory problems. Other conditions, in which inrnune suppression is desired (including, for example, organ transplantation), may also be treatable using a protein (or antagonists thereof) of the present invention. The therapeutic effects of the polypeptides or antagonists thereof on allergic reactions can be evaluated by in vivo animals models such as the cumulative contact enhancement test (Lastbom et al., Toxicology 125: 59-66, 1998), skin prick test (Hoffmann et al., Allergy 54: 446-54, 1999), guinea pig skin sensitization test (Vohr et al., Arch. Toxocol. 73: 501-9), and murine local lymph node assay (Kimber et al., J. Toxicol. Environ. Health 53: 563-79).

Using the proteins of the invention it may also be possible to modulate immune responses, in a number of ways. Down regulation may be in the form of inhibiting or blocking an immune response already in progress or may involve preventing the induction of an immune response. The functions of activated T cells may be inhibited by suppressing T cell responses or by inducing specific tolerance in T cells, or both. Immunosuppression of T cell responses is generally an active, non-antigen-specific, process which requires continuous exposure of the T cells to the suppressive agent. Tolerance, which involves inducing non-responsiveness or anergy in T cells, is distinguishable from immunosuppression in that it is generally antigen-specific and persists after exposure to the tolerizing agent has ceased. Operationally, tolerance can be demonstrated by the lack of a T cell response upon reexposure to specific antigen in the absence of the tolerizing agent.

Down regulating or preventing one or more antigen functions (including without limitation B lymphocyte antigen functions (such as, for example, B7)), e.g., preventing high level lymphokine synthesis by activated T cells, will be useful in situations of tissue, skin and organ transplantation and in graft-versus-host disease (GVHD). For example, blockage of T cell function should result in reduced tissue destruction in tissue transplantation. Typically, in tissue transplants, rejection of the transplant is initiated through its recognition as foreign by T cells, followed by an immune reaction that destroys the transplant. The administration of a therapeutic composition of the invention may prevent cytokine synthesis by immune cells, such as T cells, and thus acts as an immunosuppressant. Moreover, a lack of costimulation may also be sufficient to energize the T cells, thereby inducing tolerance in a subject. Induction of long-term tolerance by B lymphocyte antigen-blocking reagents may avoid the necessity of repeated administration of these blocking reagents. To achieve sufficient immunosuppression or tolerance in a subject, it may also be necessary to block the function of a combination of B lymphocyte antigens.

The efficacy of particular therapeutic compositions in preventing organ transplant rejection or GVHD can be assessed using animal models that are predictive of efficacy in humans. Examples of appropriate systems which can be used include allogeneic cardiac grafts in rats and xenogeneic pancreatic islet cell grafts in mice, both of which have been used to examine the immunosuppressive effects of CTLA4Ig fusion proteins in vivo as described in Lenschow et al., Science 257:789-792 (1992) and Turka et al., Proc. Natl. Acad. Sci USA, 89:11102-11105 (1992). In addition, murine models of GVHD (see Paul ed., Fundamental Immunology, Raven Press, New York, 1989, pp. 846-847) can be used to determine the effect of therapeutic compositions of the invention on the development of that disease.

Blocking antigen function may also be therapeutically useful for treating autoimmune diseases. Many autoimmune disorders are the result of inappropriate activation of T cells that are reactive against self tissue and which promote the production of cytokines and autoantibodies involved in the pathology of the diseases. Preventing the activation of autoreactive T cells may reduce or eliminate disease symptoms. Administration of reagents which block stimulation of T cells can be used to inhibit T cell activation and prevent production of autoantibodies or T cell-derived cytokines which may be involved in the disease process. Additionally, blocking reagents may induce antigen-specific tolerance of autoreactive T cells which could lead to long-term relief from the disease. The efficacy of blocking reagents in preventing or alleviating autoimmune disorders can be determined using a number of well-characterized animal models of human autoimmune diseases. Examples include murine experimental autoimmune encephalitis, systemic lupus erythmatosis in MRL/lpr/lpr mice or NZB hybrid mice, murine autoimmune collagen arthritis, diabetes mellitus in NOD mice and BB rats, and murine experimental myasthenia gravis (see Paul ed., Fundamental Immunology, Raven Press, New York, 1989, pp. 840-856).

Upregulation of an antigen function (e.g., a B lymphocyte antigen function), as a means of up regulating immune responses, may also be useful in therapy. Upregulation of immune responses may be in the form of enhancing an existing immune response or eliciting an initial immune response. For example, enhancing an immune response may be useful in cases of viral infection, including systemic viral diseases such as influenza, the common cold, and encephalitis.

Alternatively, anti-viral immune responses may be enhanced in an infected patient by removing T cells from the patient, costimulating the T cells in vitro with viral antigen-pulsed APCs either expressing a peptide of the present invention or together with a stimulatory form of a soluble peptide of the present invention and reintroducing the in vitro activated T cells into the patient. Another method of enhancing anti-viral immune responses would be to isolate infected cells from a patient, transfect them with a nucleic acid encoding a protein of the present invention as described herein such that the cells express all or a portion of the protein on their surface, and reintroduce the transfected cells into the patient. The infected cells would now be capable of delivering a costimulatory signal to, and thereby activate, T cells in vivo.

A polypeptide of the present invention may provide the necessary stimulation signal to T cells to induce a T cell mediated immune response against the transfected tumor cells. In addition, tumor cells which lack MHC class I or MHC class II molecules, or which fail to reexpress sufficient mounts of MHC class I or MHC class II molecules, can be transfected with nucleic acid encoding all or a portion of (e.g., a cytoplasmic-domain truncated portion) of an MHC class I alpha chain protein and β 2 microglobulin protein or an MHC class II alpha chain protein and an MHC class II beta chain protein to thereby express MHC class I or MHC class II proteins on the cell surface. Expression of the appropriate class I or class II MHC in conjunction with a peptide having the activity of a B lymphocyte antigen (e.g., B7-1, B7-2, B7-3) induces a T cell mediated immune response against the transfected tumor cell. Optionally, a gene encoding an antisense construct which blocks expression of an MHC class II associated protein, such as the invariant chain, can also be cotransfected with a DNA encoding a peptide having the activity of a B lymphocyte antigen to promote presentation of tumor associated antigens and induce tumor specific immunity. Thus, the induction of a T cell mediated immune response in a human subject may be sufficient to overcome tumor-specific tolerance in the subject.

The activity of a protein of the invention may, among other means, be measured by the following methods:

Suitable assays for thymocyte or splenocyte cytotoxicity include, without limitation, those described in: Current Protocols in Immunology, Ed by J. E. Coligan, A. M. Kruisbeek, D. H. Margulies, E. M. Shevach, W. Strober, Pub. Greene Publishing Associates and Wiley-Interscience (Chapter 3, In Vitro assays for Mouse Lymphocyte Function 3.1-3.19; Chapter 7, Immunologic studies in Humans); Herrmann et al., Proc. Natl. Acad. Sci. USA 78:2488-2492, 1981; Herrmann et al., J. Immunol. 128:1968-1974, 1982; Handa et al., J. Immunol. 135:1564-1572, 1985; Takai et al., I. Immunol. 137:3494-3500, 1986; Takai et al., J. Immunol. 140:508-512, 1988; Bowman et al., J. Virology 61:1992-1998; Bertagnolli et al., Cellular Immunology 133:327-341, 1991; Brown et al., J. Immunol. 153:3079-3092, 1994.

Assays for T-cell-dependent immunoglobulin responses and isotype switching (which will identify, among others, proteins that modulate T-cell dependent antibody responses and that affect Th1/Th2 profiles) include, without limitation, those described in: Maliszewski, J. Immunol. 144:3028-3033, 1990; and Assays for B cell function: In vitro antibody production, Mond, J. J. and Brunswick, M. In Current Protocols in Immunology. J. E. e.a. Coligan eds. Vol 1 pp. 3.8.1-3.8.16, John Wiley and Sons, Toronto. 1994.

Mixed lymphocyte reaction (MLR) assays (which will identify, among others, proteins that generate predominantly Th1 and CTL responses) include, without limitation, those described in: Current Protocols in Immunology, Ed by J. E. Coligan, A. M. Kruisbeek, D. H. Margulies, E. M. Shevach, W. Strober, Pub. Greene Publishing Associates and Wiley-Interscience (Chapter 3, In Vitro assays for Mouse Lymphocyte Function 3.1-3.19; Chapter 7, Immunologic studies in Humans); Takai et al., J. Immunol. 137:3494-3500, 1986; Takai et al., J. Immunol. 140:508-512, 1988; Bertagnolli et al., J. Immunol. 149:3778-3783, 1992.

Dendritic cell-dependent assays (which will identify, among others, proteins expressed by dendritic cells that activate naive T-cells) include, without limitation, those described in: Guery et al., J. Immunol. 134:536-544, 1995; Inaba et al., Journal of Experimental Medicine 173:549-559, 1991; Macatonia et al., Journal of Immunology 154:5071-5079, 1995; Porgador et al., Journal of Experimental Medicine 182:255-260, 1995; Nair et al., Journal of Virology 67:4062-4069, 1993; Huang et al., Science 264:961-965, 1994; Macatonia et al., Journal of Experimental Medicine 169:1255-1264, 1989; Bhardwaj et al., Journal of Clinical Investigation 94:797-807, 1994; and Inaba et al., Journal of Experimental Medicine 172:631-640, 1990.

Assays for lymphocyte survival/apoptosis (which will identify, among others, proteins that prevent apoptosis after superantigen induction and proteins that regulate lymphocyte homeostasis) include, without limitation, those described in: Darzynkiewicz et al., Cytometry 13:795-808, 1992; Gorczyca et al., Leukemia 7:659-670, 1993; Gorczyca et al., Cancer Research 53:1945-1951, 1993; Itoh et al., Cell 66:233-243, 1991; Zacharchuk, Journal of Immunology 145:4037-4045, 1990; Zamai et al., Cytometry 14:891-897, 1993; Gorczyca et al., International Journal of Oncology 1:639-648, 1992.

Assays for proteins that influence early steps of T-cell commitment and development include, without limitation, those described in: Antica et al., Blood 84:111-117, 1994; Fine et al., Cellular Immunology 155:111-122, 1994; Galy et al., Blood 85:2770-2778, 1995; Toki et al., Proc. Nat. Acad Sci. USA 88:7548-7551, 1991.

3.10.8 Activin/Inhibin Activity

A polypeptide of the present invention may also exhibit activin- or inhibin-related activities. A polynucleotide of the invention may encode a polypeptide exhibiting such characteristics. Inhibins are characterized by their ability to inhibit the release of follicle stimulating hormone (FSH), while activins and are characterized by their ability to stimulate the release of follicle stimulating hormone (FSH). Thus, a polypeptide of the present invention, alone or in heterodimers with a member of the inhibin family, may be useful as a contraceptive based on the ability of inhibins to decrease fertility in female mammals and decrease spermatogenesis in male mammals. Administration of sufficient amounts of other inhibins can induce infertility in these mammals. Alternatively, the polypeptide of the invention, as a homodimer or as a heterodimer with other protein subunits of the inhibin group, may be useful as a fertility inducing therapeutic, based upon the ability of activin molecules in stimulating FSH release from cells of the anterior pituitary. See, for example, U.S. Pat. No. 4,798,885. A polypeptide of the invention may also be useful for advancement of the onset of fertility in sexually immature mammals, so as to increase the lifetime reproductive performance of domestic animals such as, but not limited to, cows, sheep and pigs.

The activity of a polypeptide of the invention may, among other means, be measured by the following methods.

Assays for activin/inhibin activity include, without limitation, those described in: Vale et al., Endocrinology 91:562-572, 1972; Ling et al., Nature 321:779-782, 1986; Vale et al., Nature 321:776-779, 1986; Mason et al., Nature 318:659-663, 1985; Forage et al., Proc. Natl. Acad. Sci. USA 83:3091-3095, 1986.

3.10.9 Cremotactic/Chemokinetic Activity

A polypeptide of the present invention may be involved in chemotactic or chemokinetic activity for mammalian cells, including, for example, monocytes, fibroblasts, neutrophils, T-cells, mast cells, eosinophils, epithelial and/or endothelial cells. A polynucleotide of the invention can encode a polypeptide exhibiting such attributes. Chemotactic and chemokinetic receptor activation can be used to mobilize or attract a desired cell population to a desired site of action. Chemotactic or chemokinetic compositions (e.g. proteins, antibodies, binding partners, or modulators of the invention) provide particular advantages in treatment of wounds and other trauma to tissues, as well as in treatment of localized infections. For example, attraction of lymphocytes, monocytes or neutrophils to tumors or sites of infection may result in improved immune responses against the tumor or infecting agent.

A protein or peptide has chemotactic activity for a particular cell population if it can stimulate, directly or indirectly, the directed orientation or movement of such cell population. Preferably, the protein or peptide has the ability to directly stimulate directed movement of cells. Whether a particular protein has chemotactic activity for a population of cells can be readily determined by employing such protein or peptide in any known assay for cell chemotaxis.

Therapeutic compositions of the invention can be used in the following:

Assays for chemotactic activity (which will identify proteins that induce or prevent chemotaxis) consist of assays that measure the ability of a protein to induce the migration of cells across a membrane as well as the ability of a protein to induce the adhesion of one cell population to another cell population. Suitable assays for movement and adhesion include, without limitation, those described in: Current Protocols in Immunology, Ed by J. E. Coligan, A. M. Kruisbeek, D. H. Marguiles, E. M. Shevach, W. Strober, Pub. Greene Publishing Associates and Wiley-Interscience (Chapter 6.12, Measurernent of alpha and beta Chemokines 6.12.1-6.12.28; Taub et al. J. Clin. Invest. 95:1370-1376, 1995; Lind et al. APMIS 103:140-146, 1995; Muller et al Eur. J. Immunol. 25:1744-1748; Gruber et al. J. of Immunol. 152:5860-5867, 1994; Johnston et al. J. of Immunol. 153:1762-1768, 1994.

3.10.10 Hemostatic and Thrombolytic Activity

A polypeptide of the invention may also be involved in hemostatis or thrombolysis or thrombosis. A polynucleotide of the invention can encode a polypeptide exhibiting such attributes. Compositions may be useful in treatment of various coagulation disorders (including hereditary disorders, such as hemophilias) or to enhance coagulation and other hemostatic events in treating wounds resulting from trauma, surgery or other causes. A composition of the invention may also be useful for dissolving or inhibiting formation of thromboses and for treatment and prevention of conditions resulting therefrom (such as, for example, infarction of cardiac and central nervous system vessels (e.g., stroke).

Therapeutic compositions of the invention can be used in the following:

Assay for hemostatic and thrombolytic activity include, without limitation, those described in: Linet et al., J. Clin. Pharmacol. 26:131-140, 1986; Burdick et al., Thrombosis Res. 45:413-419, 1987; Humphrey et al., Fibrinolysis 5:71-79 (1991); Schaub, Prostaglandins 35:467-474, 1988.

3.10.11 Cancer Diagnosis and Therapy

Polypeptides of the invention may be involved in cancer cell generation, proliferation or metastasis. Detection of the presence or amount of polynucleotides or polypeptides of the invention may be useful for the diagnosis and/or prognosis of one or more types of cancer. For example, the presence or increased expression of a polynucleotide/polypeptide of the invention may indicate a hereditary risk of cancer, a precancerous condition, or an ongoing malignancy. Conversely, a defect in the gene or absence of the polypeptide may be associated with a cancer condition. Identification of single nucleotide polymorphisms associated with cancer or a predisposition to cancer may also be useful for diagnosis or prognosis.

Cancer treatments promote tumor regression by inhibiting tumor cell proliferation, inhibiting angiogenesis (growth of new blood vessels that is necessary to support tumor growth) and/or prohibiting metastasis by reducing tumor cell motility or invasiveness. Therapeutic compositions of the invention may be effective in adult and pediatric oncology including in solid phase tumors/malignancies, locally advanced tumors, human soft tissue sarcomas, metastatic cancer, including lymphatic metastases, blood cell malignancies including multiple myeloma, acute and chronic leukemnias, and lymphomas, head and neck cancers including mouth cancer, larynx cancer and thyroid cancer, lung cancers including small cell carcinoma and non-small cell cancers, breast cancers including small cell carcinoma and ductal carcinoma, gastrointestinal cancers including esophageal cancer, stomach cancer, colon cancer, colorectal cancer and polyps associated with colorectal neoplasia, pancreatic cancers, liver cancer, urologic cancers including bladder cancer and prostate cancer, malignancies of the female genital tract including ovarian carcinoma, uterine (including endometrial) cancers, and solid tumor in the ovarian follicle, kidney cancers including renal cell carcinoma, brain cancers including intrinsic brain tumors, neuroblastoma, astrocytic brain tumors, gliomas, metastatic tumor cell invasion in the central nervous system, bone cancers including osteomas, skin cancers including malignant melanoma, tumor progression of human skin keratinocytes, squamous cell carcinoma, basal cell carcinoma, hemangiopericytoma and Karposi's sarcoma.

Polypeptides, polynucleotides, or modulators of polypeptides of the invention (including inhibitors and stimulators of the biological activity of the polypeptide of the invention) may be administered to treat cancer. Therapeutic compositions can be administered in therapeutically effective dosages alone or in combination with adjuvant cancer therapy such as surgery, chemotherapy, radiotherapy, thermotherapy, and laser therapy, and may provide a beneficial effect, e.g. reducing tumor size, slowing rate of tumor growth, inhibiting metastasis, or otherwise improving overall clinical condition, without necessarily eradicating the cancer.

The composition can also be administered in therapeutically effective amounts as a portion of an anti-cancer cocktail. An anti-cancer cocktail is a mixture of the polypeptide or modulator of the invention with one or more anti-cancer drugs in addition to a pharmaceutically acceptable carrier for delivery. The use of anti-cancer cocktails as a cancer treatment is routine. Anti-cancer drugs that are well known in the art and can be used as a treatment in combination with the polypeptide or modulator of the invention include: Actinomycin D, Aminoglutethimide, Asparaginase, Bleomycin, Busulfan, Carboplatin, Carmustine, Chlorambucil, Cisplatin (cis-DDP), Cyclophosphamide, Cytarabine HCl (Cytosine arabinoside), Dacarbazine, Dactinomycin, Daunorubicin HCl, Doxorubicin HCl, Estramustine phosphate sodium, Etoposide (V16-213), Floxuridine, 5-Fluorouracil (5-Fu), Flutamide, Hydroxyurea (hydroxycarbamide), Ifosfamide, Interferon Alpha-2a, Interferon Alpha-2b, Leuprolide acetate (LHRH-releasing factor analog), Lomustine, Mechiorethamnine HCl (nitrogen mustard), Melphalan, Mercaptopurine, Mesna, Methotrexate (MTX), Mitomycin, Mitoxantrone HCl, Octreotide, Plicamycin, Procarbazine HCl, Streptozocin, Tamoxifen citrate, Thioguanine, Thiotepa, Vinblastine sulfate, Vincristine sulfate, Ainsacrinc, Azacitidine, Hexamethylnelamine, Interleukin-2, Mitoguazone, Pentostatin, Semustine, Teniposide, and Vindesine sulfate.

In addition, therapeutic compositions of the invention may be used for prophylactic treatment of cancer. There are hereditary conditions and/or environmental situations (e.g. exposure to carcinogens) known in the art that predispose an individual to developing cancers. Under these circumstances, it may be beneficial to treat these individuals with therapeutically effective doses of the polypeptide of the invention to reduce the risk of developing cancers.

In vitro models can be used to determine the effective doses of the polypeptide of the invention as a potential cancer treatment. These in vitro models include proliferation assays of cultured tumor cells, growth of cultured tumor cells in soft agar (see Freshney, (1987) Culture of Animal Cells: A Manual of Basic Technique, Wily-Liss, New York, N.Y. Ch 18 and Ch 21), tumor systems in nude mice as described in Giovanella et al., J. Natl. Can. Inst., 52: 921-30 (1974), mobility and invasive potential of tumor cells in Boyden Chamber assays as described in Pilkington et al., Anticancer Res., 17: 4107-9 (1997), and angiogenesis assays such as induction of vascularization of the chick chorioallantoic membrane or induction of vascular endothelial cell migration as described in Ribatta et al., Intl. J. Dev. Biol., 40: 1189-97 (1999) and Li et al., Clin. Exp. Metastasis, 17:423-9 (1999), respectively. Suitable tumor cells lines are available, e.g. from American Type Tissue Culture Collection catalogs.

3.10.12 Receptor/Ligand Activity

A polypeptide of the present invention may also demonstrate activity as receptor, receptor ligand or inhibitor or agonist of receptor/ligand interactions. A polynucleotide of the invention can encode a polypeptide exhibiting such characteristics. Examples of such receptors and ligands include, without limitation, cytokine receptors and their ligands, receptor kinases and their ligands, receptor phosphatases and their ligands, receptors involved in cell-cell interactions and their ligands (including without limitation, cellular adhesion molecules (such as selectins, integrins and their ligands) and receptor/ligand pairs involved in antigen presentation, antigen recognition and development of cellular and humoral immune responses. Receptors and ligands are also useful for screening of potential peptide or small molecule inhibitors of the relevant receptor/ligand interaction. A protein of the present invention (including, without limitation, fragments of receptors and ligands) may themselves be useful as inhibitors of receptor/ligand interactions.

The activity of a polypeptide of the invention may, among other means, be measured by the following methods:

Suitable assays for receptor-ligand activity include without limitation those described in: Current Protocols in Immunology, Ed by J. E. Coligan, A. M. Kruisbeek, D. H. Margulies, E. M. Shevach, W. Strober, Pub. Greene Publishing Associates and Wiley-Interscience (Chapter 7.28, Measurement of Cellular Adhesion under static conditions 7.28.1-7.28.22), Takai et al., Proc. Natl. Acad. Sci. USA 84:6864-6868, 1987; Bierer et al., J. Exp. Med. 168:1145-1156, 1988; Rosenstein et al., J. Exp. Med. 169:149-160 1989; Stoltenborg et al., J. Immunol. Methods 175:59-68, 1994; Stitt et al., Cell 80:661-670, 1995.

By way of example, the polypeptides of the invention may be used as a receptor for a ligand(s) thereby transmitting the biological activity of that ligand(s). Ligands may be identified through binding assays, affinity chromatography, dihybrid screening assays, BIAcore assays, gel overlay assays, or other methods known in the art.

Studies characterizing drugs or proteins as agonist or antagonist or partial agonists or a partial antagonist require the use of other proteins as competing ligands. The polypeptides of the present invention or ligand(s) thereof may be labeled by being coupled to radioisotopes, colorimetric molecules or a toxin molecules by conventional methods. (“Guide to Protein Purification” Murray P. Deutscher (ed) Methods in Enzymology Vol. 182 (1990) Academic Press, Inc. San Diego). Examples of radioisotopes include, but are not limited to, tritium and carbon-14. Examples of calorimetric molecules include, but are not limited to, fluorescent molecules such as fluorescamine, or rhodamine or other colorimetric molecules. Examples of toxins include, but are not limited, to ricin.

3.10.13 Drug Screening

This invention is particularly useful for screening chemical compounds by using the novel polypeptides or binding fragments thereof in any of a variety of drug screening techniques. The polypeptides or fragments employed in such a test may either be free in solution, affixed to a solid support, borne on a cell surface or located intracellularly. One method of drug screening utilizes eukaryotic or prokaryotic host cells which are stably transformed with recombinant nucleic acids expressing the polypeptide or a fragment thereof. Drugs are screened against such transformed cells in competitive binding assays. Such cells, either in viable or fixed form, can be used for standard binding assays. One may measure, for example, the formation of complexes between polypeptides of the invention or fragments and the agent being tested or examine the diminution in complex formation between the novel polypeptides and an appropriate cell line, which are well known in the art.

Sources for test compounds that may be screened for ability to bind to or modulate (i.e., increase or decrease) the activity of polypeptides of the invention include (1) inorganic and organic chemical libraries, (2) natural product libraries, and (3) combinatorial libraries comprised of either random or mimetic peptides, oligonucleotides or organic molecules.

Chemical libraries may be readily synthesized or purchased from a number of commercial sources, and may include structural analogs of known compounds or compounds that are identified as “hits” or “leads” via natural product screening.

The sources of natural product libraries are microorganisms (including bacteria and fungi), animals, plants or other vegetation, or marine organisms, and libraries of mixtures for screening may be created by: (1) fermentation and extraction of broths from soil, plant or marine microorganisms or (2) extraction of the organisms themselves. Natural product libraries include polyketides, non-ribosomal peptides, and (non-naturally occurring) variants thereof. For a review, see Science 282:63-68 (1998).

Combinatorial libraries are composed of large numbers of peptides, oligonucleotides or organic compounds and can be readily prepared by traditional automated synthesis methods, PCR, cloning or proprietary synthetic methods. Of particular interest are peptide and oligonucleotide combinatorial libraries. Still other libraries of interest include peptide, protein, peptidomimetic, multiparallel synthetic collection, recombinatorial, and polypeptide libraries. For a review of combinatorial chemistry and libraries created therefrom, see Myers, Curr. Opin. Biotechnol . 8:701-707 (1997). For reviews and examples of peptidomimetic libraries, see Al-Obeidi et al., Mol. Biotechnol , 9(3):205-23 (1998); Hrubyet al., Curr Opin Chem Biol, 1(1):114-19(1997); Domer et al., Bioorg Med Chem , 4(5):709-15 (1996) (alkylated dipeptides).

Identification of modulators through use of the various libraries described herein permits modification of the candidate “hit” (or “lead”) to optimize the capacity of the “hit” to bind a polypeptide of the invention. The molecules identified in the binding assay are then tested for antagonist or agonist activity in in vivo tissue culture or animal models that are well known in the art. In brief, the molecules are titrated into a plurality of cell cultures or animals and then tested for either cell/animal death or prolonged survival of the animal/cells.

The binding molecules thus identified may be complexed with toxins, e.g., ricin or cholera, or with other compounds that are toxic to cells such as radioisotopes. The toxin-binding molecule complex is then targeted to a tumor or other cell by the specificity of the binding molecule for a polypeptide of the invention. Alternatively, the binding molecules may be complexed with imaging agents for targeting and imaging purposes.

3.10.14 Assay for Receptor Activity

The invention also provides methods to detect specific binding of a polypeptide e.g. a ligand or a receptor. The art provides numerous assays particularly useful for identifying previously unknown binding partners for receptor polypeptides of the invention. For example, expression cloning using mammalian or bacterial cells, or dihybrid screening assays can be used to identify polynucleotides encoding binding partners. As another example, affinity chromatography with the appropriate immobilized polypeptide of the invention can be used to isolate polypeptides that recognize and bind polypeptides of the invention. There are a number of different libraries used for the identification of compounds, and in particular small molecules, that modulate (i.e., increase or decrease) biological activity of a polypeptide of the invention. Ligands for receptor polypeptides of the invention can also be identified by adding exogenous ligands, or cocktails of ligands to two cells populations that are genetically identical except for the expression of the receptor of the invention: one cell population expresses the receptor of the invention whereas the other does not. The response of the two cell populations to the addition of ligands(s) are then compared. Alternatively, an expression library can be co-expressed with the polypeptide of the invention in cells and assayed for an autocrine response to identify potential ligand(s). As still another example, BIAcore assays, gel overlay assays, or other methods known in the art can be used to identify binding partner polypeptides, including, (1) organic and inorganic chemical libraries, (2) natural product libraries, and (3) combinatorial libraries comprised of random peptides, oligonuclcotides or organic molecules.

The role of downstream intracellular signaling molecules in the signaling cascade of the polypeptide of the invention can be determined. For example, a chimeric protein in which the cytoplasmic domain of the polypeptide of the invention is fused to the extracellular portion of a protein, whose ligand has been identified, is produced in a host cell. The cell is then incubated with the ligand specific for the extracellular portion of the chimeric protein, thereby activating the chimeric receptor. Known downstream proteins involved in intracellular signaling can then be assayed for expected modifications i.e. phosphorylation. Other methods known to those in the art can also be used to identify signaling molecules involved in receptor activity.

3.10.15 Anti-inflammatory Activity

Compositions of the present invention may also exhibit anti-inflammatory activity. The anti-inflammatory activity may be achieved by providing a stimulus to cells involved in the inflammatory response, by inhibiting or promoting cell-cell interactions (such as, for example, cell adhesion), by inhibiting or promoting chemotaxis of cells involved in the inflammatory process, inhibiting or promoting cell extravasation, or by stimulating or suppressing production of other factors which more directly inhibit or promote an inflammatory response. Compositions with such activities can be used to treat inflammatory conditions including chronic or acute conditions), including without limitation intimation associated with infection (such as septic shock, sepsis or systemic inflammatory response syndrome (SIRS)), ischemia-reperfusion injury, endotoxin lethality, arthritis, complement-mediated hyperacute rejection, nephritis, cytokine or chemokine-induced lung injury, inflammatory bowel disease, Crohn's disease or resulting from over production of cytokines such as TNF or IL-1. Compositions of the invention may also be useful to treat anaphylaxis and hypersensitivity to an antigenic substance or material. Compositions of this invention may be utilized to prevent or treat conditions such as, but not limited to, sepsis, acute pancreatitis, endotoxin shock, cytokine induced shock, rheumatoid arthritis, chronic inflammatory arthritis, pancreatic cell damage from diabetes mellitus type 1, graft versus host disease, inflammatory bowel disease, inflamation associated with pulmonary disease, other autoimmune disease or inflammatory disease, an antiproliferative agent such as for acute or chronic mylegenous leukemia or in the prevention of premature labor secondary to intrauterine infections.

3.10.16 Leukemias

Leukemias and related disorders may be treated or prevented by administration of a therapeutic that promotes or inhibits function of the polynucleotides and/or polypeptides of the invention. Such leukemias and related disorders include but are not limited to acute leukemia, acute lymphocytic leukemia, acute myelocytic leukemia, myeloblastic, promyclocytic, myelomonocytic, monocytic, erythroleukemia, chronic leukemia, chronic myelocytic (granulocytic) leukemia and chronic lymphocytic leukemia (for a review of such disorders, see Fishman et al., 1985, Medicine, 2d Ed., J. B. Lippincott Co., Philadelphia).

3.10.17 Nervous System Disorders

Nervous system disorders, involving cell types which can be tested for efficacy of intervention with compounds that modulate the activity of the polynucleotides and/or polypeptides of the invention, and which can be treated upon thus observing an indication of therapeutic utility, include but are not limited to nervous system injuries, and diseases or disorders which result in either a disconnection of axons, a diminution or degeneration of neurons, or demyelination. Nervous system lesions which may be treated in a patient (including human and non-human mammalian patients) according to the invention include but are not limited to the following lesions of either the central (including spinal cord, brain) or peripheral nervous systems:

(i) traumatic lesions, including lesions caused by physical injury or associated with surgery, for example, lesions which sever a portion of the nervous system, or compression injuries;

(ii) ischemnic lesions, in which a lack of oxygen in a portion of the nervous system results in neuronal injury or death, including cerebral infarction or ischemia, or spinal cord infarction or ischemia;

(iii) infectious lesions, in which a portion of the nervous system is destroyed or injured as a result of infection, for example, by an abscess or associated with infection by human immunodeficiency virus, herpes zoster, or herpes simplex virus or with Lymc disease, tuberculosis, syphilis;

(iv) degenerative lesions, in which a portion of the nervous system is destroyed or injured as a result of a degenerative process including but not limited to degeneration associated with Parkinson's disease, Alzheimer's disease, Huntington's chorea, or amyotrophic lateral sclerosis;

(v) lesions associated with nutritional diseases or disorders, in which a portion of the nervous system is destroyed or injured by a nutritional disorder or disorder of metabolism including but not limited to, vitamin B12 deficiency, folic acid deficiency, Wemicke disease, tobacco-alcohol amblyopia, Marchiafava-Bignami disease (primary degeneration of the corpus callosum), and alcoholic cerebellar degeneration;

(vi) neurological lesions associated with systemic diseases including but not limited to diabetes (diabetic neuropathy, Bell's palsy), systemic lupus erythematosus, carcinoma, or sarcoidosis;

(vii) lesions caused by toxic substances including alcohol, lead, or particular neurotoxins; and

(viii) demyelinated lesions in which a portion of the nervous system is destroyed or injured by a demyelinating disease including but not limited to multiple sclerosis, human immunodeficiency virus-associated myelopathy, transverse myelopathy or various etiologies, progressive multifocal leukoencephalopathy, and central pontine myelinolysis.

Therapeutics which are useful according to the invention for treatment of a nervous system disorder may be selected by testing for biological activity in promoting the survival or differentiation of neurons. For example, and not by way of limitation, therapeutics which elicit any of the following effects may be useful according to the invention:

(i) increased survival time of neurons in culture;

(ii) increased sprouting of neurons in culture or in vivo;

(iii) increased production of a neuron-associated molecule in culture or in vivo, e.g., choline acetyltransferase or acetylcholinesterase with respect to motor neurons; or

(iv) decreased symptoms of neuron dysfimction in vivo.

Such effects may be measured by any method known in the art. In preferred, non-limiting embodiments, increased survival of neurons may be measured by the method set forth in Arakawa et al. (1990, J. Neurosci. 10:3507-3515); increased sprouting of neurons may be detected by methods set forth in Pestronk et al. (1980, Exp. Neurol. 70:65-82) or Brown et al. (1981, Ann. Rev. Neurosci. 4:1742); increased production of neuron-associated molecules may be measured by bioassay, enzymatic assay, antibody binding, Northern blot assay, etc., depending on the molecule to be measured; and motor neuron dysfunction may be measured by assessing the physical manifestation of motor neuron disorder, e.g., weakness, motor neuron conduction velocity, or functional disability.

In specific embodiments, motor neuron disorders that may be treated according to the invention include but are not limited to disorders such as infarction, infection, exposure to toxin, trauma, surgical damage, degenerative disease or malignancy that may affect motor neurons as well as other components of the nervous system, as well as disorders that selectively affect neurons such as amyotrophic lateral sclerosis, and including but not limited to progressive spinal muscular atrophy, progressive bulbar palsy, primary lateral sclerosis, infantile and juvenile muscular atrophy, progressive bulbar paralysis of childhood (Fazio-Londe syndrome), poliomyelitis and the post polio syndrome, and Hereditary Motorsensory Neuropathy (Charcot-Marie-Tooth Disease).

3.10.18 Other Activities

A polypeptide of the invention may also exhibit one or more of the following additional activities or effects: inhibiting the growth, infection or function of, or killing, infectious agents, including, without limitation, bacteria, viruses, fuingi and other parasites; effecting (suppressing or enhancing) bodily characteristics, including, without limitation, height, weight, hair color, eye color, skin, fat to lean ratio or other tissue pigrnentation, or organ or body part size or shape (such as, for example, breast augmentation or diminution, change in bone form or shape); effecting biorhythms or circadian cycles or rhythms; effecting the fertility of male or female subjects; effecting the metabolism, catabolism, anabolism, processing, utilization, storage or elimination of dietary fat, lipid, protein, carbohydrate, vitamins, minerals, co-factors or other nutritional factors or component(s); effecting behavioral characteristics, including, without limitation, appetite, libido, stress, cognition (including cognitive disorders), depression (including depressive disorders) and violent behaviors; providing analgesic effects or other pain reducing effects; promoting differentiation and growth of embryonic stem cells in lineages other than hematopoietic lineages; hormonal or endocrine activity; in the case of enzymes, correcting deficiencies of the enzyme and treating deficiency-related diseases; treatment of hyperproliferative disorders (such as, for example, psoriasis); immunoglobulin-like activity (such as, for example, the ability to bind antigens or complement); and the ability to act as an antigen in a vaccine composition to raise an immune response against such protein or another material or entity which is cross-reactive with such protein.

3.10.19 Identification of Polymorphisms

The demonstration of polymorphisms makes possible the identification of such polymorphisms in human subjects and the pharmacogenetic use of this information for diagnosis and treatment. Such polymorphisms may be associated with, e.g., differential predisposition or susceptibility to various disease states (such as disorders involving inflammation or immune response) or a differential response to drug admninistration, and this genetic information can be used to tailor preventive or therapeutic treatment appropriately. For example, the existence of a polymorphism associated with a predisposition to inflammation or autoimmune disease makes possible the diagnosis of this condition in humans by identifying the presence of the polymorphism.

Polymorphisms can be identified in a variety of ways known in the art which all generally involve obtaining a sample from a patient, analyzing DNA from the sample, optionally involving isolation or amplification of the DNA, and identifying the presence of the polymorphism in the DNA. For example, PCR may be used to amplify an appropriate fragment of genomic DNA which may then be sequenced. Alternatively, the DNA may be subjected to allele-specific oligonucleotide hybridization (in which appropriate oligonucleotides are hybridized to the DNA under conditions permitting detection of a single base mismatch) or to a single nucleotide extension assay (in which an oligonucleotide that hybridizes immediately adjacent to the position of the polymorphism is extended with one or more labeled nucleotides). In addition, traditional restriction fragment length polymorphism analysis (using restriction enzymes that provide differential digestion of the genomic DNA depending on the presence or absence of the polymorphism) may be performed. Arrays with nucleotide sequences of the present invention can be used to detect polymorphisms. The array can comprise modified nucleotide sequences of the present invention in order to detect the nucleotide sequences of the present invention. In the alternative, any one of the nucleotide sequences of the present invention can be placed on the array to detect changes from those sequences.

Alternatively a polymorphism resulting in a change in the amino acid sequence could also be detected by detecting a corresponding change in amino acid sequence of the protein, e.g., by an antibody specific to the variant sequence. 3.10.20 Arthritis and Inflammation

The immunosuppressive effects of the compositions of the invention against rheumatoid arthritis is determined in an experimental animal model system. The experimental model system is adjuvant induced arthritis in rats, and the protocol is described by J. Holoshitz, et at., 1983, Science, 219:56, or by B. Waksman et al., 1963, Int. Arch. Allergy Appl. Immunol., 23:129. Induction of the disease can be caused by a single injection, generally intradermally, of a suspension of killed Mycobacterium tuberculosis in complete Freund's adjuvant (CFA). The route of injection can vary, but rats may be injected at the base of the tail with an adjuvant mixture. The polypeptide is administered in phosphate buffered solution (PBS) at a dose of about 1-5 mg/kg. The control consists of administering PBS only.

The procedure for testing the effects of the test compound would consist of intradermally injecting killed Mycobacterium tuberculosis in CFA followed by immediately administering the test compound and subsequent treatment every other day until day 24. At 14, 15, 18, 20, 22, and 24 days after injection of Mycobacterium CFA, an overall arthritis score may be obtained as described by J. Holoskitz above. An analysis of the data would reveal that the test compound would have a dramatic affect on the swelling of the joints as measured by a decrease of the arthritis score.

3.11 Therapeutic Methods

The compositions (including polypeptide fragments, analogs, variants and antibodies or other binding partners or modulators including antisense polynucleotides) of the invention have numerous applications in a variety of therapeutic methods. Examples of therapeutic applications include, but are not limited to, those exemplified herein.

3.11.1 Example

One embodiment of the invention is the administration of an effective amount of the polypeptides or other composition of the invention to individuals affected by a disease or disorder that can be modulated by regulating the peptides of the invention. While the mode of administration is not particularly important, parenteral administration is preferred. An exemplary mode of administration is to deliver an intravenous bolus. The dosage of the polypeptides or other composition of the invention will normally be determined by the prescribing physician. It is to be expected that the dosage will vary according to the age, weight, condition and response of the individual patient. Typically, the amount of polypeptide administered per dose will be in the range of about 0.01 μg/kg to 100 mg/kg of body weight, with the preferred dose being about 0.1 μg/kg to 10 mg/kg of patient body weight. For parenteral administration, polypeptides of the invention will be formulated in an injectable form combined with a pharmaceutically acceptable parenteral vehicle. Such vehicles are well known in the art and examples include water, saline, Ringer's solution, dextrose solution, and solutions consisting of small amounts of the human serum albumin. The vehicle may contain minor amounts of additives that maintain the isotonicity and stability of the polypeptide or other active ingredient. The preparation of such solutions is within the slill of the art.

3.12 Pharmaceutical Formulations and Routes of Administration

A protein or other composition of the present invention (from whatever source derived, including without limitation from recombinant and non-recombinant sources and including antibodies and other binding partners of the polypeptides of the invention) may be administered to a patient in need, by itself, or in pharmaceutical compositions where it is mixed with suitable carriers or excipient(s) at doses to treat or ameliorate a variety of disorders. Such a composition may optionally contain (in addition to protein or other active ingredient and a carrier) diluents, fillers, salts, buffers, stabilizers, solubilizers, and other materials well known in the art. The term “pharmaceutically acceptable” means a non-toxic material that does not interfere with the effectiveness of the biological activity of the active ingredient(s). The characteristics of the carrier will depend on the route of administration. The pharmaceutical composition of the invention may also contain cytokines, lymphokines, or other hematopoietic factors such as M-CSF, GM-CSF, TNF, IL-1, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12, IL-13, IL-14, IL-15, IFN, TNF0, TNF1, TNF2, G-CSF, Meg-CSF, thrombopoietin, stem cell factor, and erythropoietin. In further compositions, proteins of the invention may be combined with other agents beneficial to the treatment of the disease or disorder in question. These agents include various growth factors such as epidermal growth factor (EGF), platelet-derived growth factor (PDGF), transforming growth factors (TGF-α and TGF-β), insulin-like growth factor (IGF), as well as cytokines described herein.

The pharmaceutical composition may further contain other agents which either enhance the activity of the protein or other active ingredient or complement its activity or use in treatment. Such additional factors and/or agents may be included in the pharmaceutical composition to produce a synergistic effect with protein or other active ingredient of the invention, or to minimize side effects. Conversely, protein or other active ingredient of the present invention may be included in formulations of the particular clotting factor, cytokine, lymphokine, other hematopoietic factor, thrombolytic or anti-thrombotic factor, or anti-inflammatory agent to minimize side effects of the clotting factor, cytokine, lymphokine, other hematopoietic factor, thrombolytic or anti-thrombotic factor, or anti-inflammatory agent (such as IL-1Ra, IL-1 Hy1, IL-1 Hy2, anti-TNF, corticosteroids, immunosuppressive agents). A protein of the present invention may be active in multimers (e.g., heterodimers or homodimers) or complexes with itself or other proteins. As a result, pharmaceutical compositions of the invention may comprise a protein of the invention in such multimeric or complexed form.

As an alternative to being included in a pharmaceutical composition of the invention including a first protein, a second protein or a therapeutic agent may be concurrently administered with the first protein (e.g., at the same time, or at differing times provided that therapeutic concentrations of the combination of agents is achieved at the treatment site). Techniques for formulation and administration of the compounds of the instant application may be found in “Remington's Pharmaceutical Sciences,” Mack Publishing Co., Easton, Pa., latest edition. A therapeutically effective dose further refers to that amount of the compound sufficient to result in amelioration of symptoms, e.g., treatment, healing, prevention or amelioration of the relevant medical condition, or an increase in rate of treatment, healing, prevention or amelioration of such conditions. When applied to an individual active ingredient, administered alone, a therapeutically effective dose refers to that ingredient alone. When applied to a combination, a therapeutically effective dose refers to combined amounts of the active ingredients that result in the therapeutic effect, whether administered in combination, serially or simultaneously.

In practicing the method of treatment or use of the present invention, a therapeutically effective amount of protein or other active ingredient of the present invention is administered to a mammal having a condition to be treated. Protein or other active ingredient of the present invention may be administered in accordance with the method of the invention either alone or in combination with other therapies such as treatments employing cytokines, lymphokines or other hematopoietic factors. When co-administered with one or more cytokines, lymphokines or other hematopoiefic factors, protein or other active ingredient of the present invention may be administered either simultaneously with the cytokine(s), lymphokine(s), other hematopoietic factor(s), thrombolytic or anti-thrombotic factors, or sequentially. If administered sequentially, the attending physician will decide on the appropriate sequence of administering protein or other active ingredient of the present invention in combination with cytokine(s), lymphokine(s), other hematopoietic factor(s), thrombolytic or anti-thrombotic factors.

3.12.1 Routes of Administration

Suitable routes of administration may, for example, include oral, rectal, transmucosal, or intestinal administration; parenteral delivery, including intramuscular, subcutaneous, intramedullary injections, as well as intrathecal, direct intraventricular, intravenous, intraperitoneal, intranasal, or intraocular injections. Administration of protein or other active ingredient of the present invention used in the pharmaceutical composition or to practice the method of the present invention can be carried out in a variety of conventional ways, such as oral ingestion, inhalation, topical application or cutaneous, subcutaneous, intraperitoneal, parenteral or intravenous injection. Intravenous administration to the patient is preferred.

Alternately, one may administer the compound in a local rather than systemic manner, for example, via injection of the compound directly into a arthritic joints or in fibrotic tissue, often in a depot or sustained release formulation. In order to prevent the scarring process frequently occurring as complication of glaucoma surgery, the compounds may be administered topically, for example, as eye drops. Furthermore, one may administer the drug in a targeted drug delivery system, for example, in a liposome coated with a specific antibody, targeting, for example, arthritic or fibrotic tissue. The liposomes will be targeted to and taken up selectively by the afflicted tissue.

The polypeptides of the invention are administered by any route that delivers an effective dosage to the desired site of action. The determination of a suitable route of administration and an effective dosage for a particular indication is within the level of skill in the art. Preferably for wound treatment, one administers the therapeutic compound directly to the site. Suitable dosage ranges for the polypeptides of the invention can be extrapolated from these dosages or from similar studies in appropriate animal models. Dosages can then be adjusted as necessary by the clinician to provide maximal therapeutic benefit.

3.12.2 Compositions/Formulations

Pharmaceutical compositions for use in accordance with the present invention thus may be formulated in a conventional manner using one or more physiologically acceptable carriers comprising excipients and auxiliaries which facilitate processing of the active compounds into preparations which can be used pharmaceutically. These pharmaceutical compositions may be manufactured in a manner that is itself known, e.g., by means of conventional mixing, dissolving, granulating, dragee-making, levigating, emulsifying, encapsulating, entrapping or lyophilizing processes. Proper formulation is dependent upon the route of administration chosen. When a therapeutically effective amount of protein or other active ingredient of the present invention is administered orally, protein or other active ingredient of the present invention will be in the form of a tablet, capsule, powder, solution or elixir. When administered in tablet forrm, the pharmaceutical composition of the invention may additionally contain a solid carrier such as a gelatin or an adjuvant. The tablet, capsule, and powder contain from about 5 to 95% protein or other active ingredient of the present invention, and preferably from about 25 to 90% protein or other active ingredient of the present invention. When administered in liquid form, a liquid carrier such as water, petroleum, oils of animal or plant origin such as peanut oil, mineral oil, soybean oil, or sesame oil, or synthetic oils may be added. The liquid form of the pharmaceutical composition may bather contain physiological saline solution, dextrose or other saccharide solution, or glycols such as ethylene glycol, propylene glycol or polyethylene glycol. When administered in liquid form, the pharmaceutical composition contains from about 0.5 to 90% by weight of protein or other active ingredient of the present invention, and preferably from about 1 to 50% protein or other active ingredient of the present invention.

When a therapeutically effective amount of protein or other active ingredient of the present invention is administered by intravenous, cutaneous or subcutaneous injection, protein or other active ingredient of the present invention will be in the form of a pyrogen-free, parenterally acceptable aqueous solution. The preparation of such parenterally acceptable protein or other active ingredient solutions, having due regard to pH, isotonicity, stability, and the like, is within the skill in the art. A preferred pharmaceutical composition for intravenous, cutaneous, or subcutaneous injection should contain, in addition to protein or other active ingredient of the present invention, an isotonic vehicle such as Sodium Chloride Injection, Ringer's Injection, Dextrose Injection, Dextrose and Sodium Chloride Injection, Lactated Ringer's Injection, or other vehicle as known in the art. The pharmaceutical composition of the present invention may also contain stabilizers, preservatives, buffers, antioxidants, or other additives known to those of skill in the art. For injection, the agents of the invention may be formulated in aqueous solutions, preferably in physiologically compatible buffers such as Hanks's solution, Ringer's solution, or physiological saline buffer. For tarsmucosal administration, penetrants appropriate to the barrier to be permeated are used in the formulation. Such penetrants are generally known in the art.

For oral administration, the compounds can be formulated readily by combining the active compounds with pharmaceutically acceptable carriers well known in the art. Such carriers enable the compounds of the invention to be formulated as tablets, pills, dragees, capsules, liquids, gels, syrups, slurries, suspensions and the like, for oral ingestion by a patient to be treated. Pharmaceutical preparations for oral use can be obtained from a solid excipient, optionally grinding a resulting mixture, and processing the mixture of granules, after adding suitable auxiliaries, if desired, to obtain tablets or dragee cores. Suitable excipients are, in particular, fillers such as sugars, including lactose, sucrose, mannitol, or sorbitol; cellulose preparations such as, for example, maize starch, wheat starch, rice starch, potato starch, gelatin, gum tragacanth, methyl cellulose, hydroxypropylmethyl-cellulose, sodium carboxymethylcellulose, and/or polyvinylpyrrolidone (PVP). If desired, disintegrating agents may be added, such as the cross-linked polyvinyl pyrrolidone, agar, or alginic acid or a salt thereof such as sodium alginate. Dragee cores are provided with suitable coatings. For this purpose, concentrated sugar solutions may be used, which may optionally contain gum arabic, talc, polyvinyl pyrrolidone, carbopol gel, polyethylene glycol, and/or titanium dioxide, lacquer solutions, and suitable organic solvents or solvent mixtures. Dyestuffs or pigments may be added to the tablets or dragee coatings for identification or to characterize different combinations of active compound doses.

Pharmaceutical preparations which can be used orally include push-fit capsules made of gelatin, as well as soft, sealed capsules made of gelatin and a plasticizer, such as glycerol or sorbitol. The push-fit capsules can contain the active ingredients in admixture with filler such as lactose, binders such as starches, and/or lubricants such as talc or magnesium stearate and, optionally, stabilizers. In soft capsules, the active compounds may be dissolved or suspended in suitable liquids, such as fatty oils, liquid paraffin, or liquid polyethylene glycols. In addition, stabilizers may be added. All formulations for oral administration should be in dosages suitable for such administration. For buccal administration, the compositions may take the form of tablets or lozenges formulated in conventional manner.

For administration by inhalation, the compounds for use according to the present invention are conveniently delivered in the form of an aerosol spray presentation from pressurized packs or a nebuliser, with the use of a suitable propellant, e.g., dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas. In the case of a pressurized aerosol the dosage unit may be determined by providing a valve to deliver a metered amount. Capsules and cartridges of, e.g., gelatin for use in an inhaler or insufflator may be formulated containing a powder mix of the compound and a suitable powder base such as lactose or starch. The compounds may be formulated for parenteral administration by injection, e.g., by bolus injection or continuous infusion. Formulations for injection may be presented in unit dosage form, e.g., in ampules or in multi-dose containers, with an added preservative. The compositions may take such forms as suspensions, solutions or emulsions in oily or aqueous vehicles, and may contain formulatory agents such as suspending, stabilizing and/or dispersing agents.

Pharmaceutical formulations for parenteral administration include aqueous solutions of the active compounds in water-soluble form. Additionally, suspensions of the active compounds may be prepared as appropriate oily injection suspensions. Suitable lipophilic solvents or vehicles include fatty oils such as sesame oil, or synthetic fatty acid esters, such as ethyl oleate or triglycerides, or liposomes. Aqueous injection suspensions may contain substances which increase the viscosity of the suspension, such as sodium carboxymethyl cellulose, sorbitol, or dextran. Optionally, the suspension may also contain suitable stabilizers or agents which increase the solubility of the compounds to allow for the preparation of highly concentrated solutions. Alternatively, the active ingredient may be in powder form for constitution with a suitable vehicle, e.g., sterile pyrogen-free water, before use.

The compounds may also be formulated in rectal compositions such as suppositories or retention enemas, e.g., containing conventional suppository bases such as cocoa butter or other glycerides. In addition to the formulations described previously, the compounds may also be formulated as a depot preparation. Such long acting formulations may be administered by implantation (for example subcutaneously or intramuscularly) or by intramuscular injection. Thus, for example, the compounds may be formulated with suitable polymeric or hydrophobic materials (for example as an emulsion in an acceptable oil) or ion exchange resins, or as sparingly soluble derivatives, for example, as a sparingly soluble salt.

A pharmaceutical carrier for the hydrophobic compounds of the invention is a co-solvent system comprising benzyl alcohol, a nonpolar surfactant, a water-miscible organic polymer, and an aqueous phase. The co-solvent system may be the VPD co-solvent system. VPD is a solution of 3% w/v benzyl alcohol, 8% w/v of the nonpolar surfactant polysorbate 80, and 65% w/v polyethylene glycol 300, made up to volume in absolute ethanol. The VPD co-solvent system (VPD:5W) consists of VPD diluted 1:1 with a 5% dextrose in water solution. This co-solvent system dissolves hydrophobic compounds well, and itself produces low toxicity upon systemic administration. Naturally, the proportions of a co-solvent system may be varied considerably without destroying its solubility and toxicity characteristics. Furthermnore, the identity of the co-solvent components may be varied: for example, other low-toxicity nonpolar surfactants may be used instead of polysorbate 80; the fraction size of polyethylene glycol may be varied; other biocompatible polymers may replace polyethylene glycol, e.g. polyvinyl pyrrolidone; and other sugars or polysaccharides may substitute for dextrose. Alternatively, other delivery systems for hydrophobic pharmaceutical compounds may be employed. Liposomes and emulsions are well known examples of delivery vehicles or carriers for hydrophobic drugs. Certain organic solvents such as dimethylsulfoxide also may be employed, although usually at the cost of greater toxicity. Additionally, the compounds may be delivered using a sustained-release system, such as semipermeable matrices of solid hydrophobic polymers containing the therapeutic agent. Various types of sustained-release materials have been established and are well known by those skilled in the art. Sustained-release capsules may, depending on their chemical nature, release the compounds for a few weeks up to over 100 days. Depending on the chemical nature and the biological stability of the therapeutic reagent, additional strategies for protein or other active ingredient stabilization may be employed.

The pharmaceutical compositions also may comprise suitable solid or gel phase carriers or excipients. Examples of such carriers or excipients include but are not limited to calcium carbonate, calcium phosphate, various sugars, starches, cellulose derivatives, gelatin, and polymers such as polyethylene glycols. Many of the active ingredients of the invention may be provided as salts with pharmaceutically compatible counter ions. Such pharmaceutically acceptable base addition salts are those salts which retain the biological effectiveness and properties of the free acids and which are obtained by reaction with inorganic or organic bases such as sodium hydroxide, magnesium hydroxide, ammonia, trialkylamine, dialkylamine, monoalkylamine, dibasic amino acids, sodium acetate, potassium benzoate, triethanol amine and the like.

The pharmaceutical composition of the invention may be in the form of a complex of the protein(s) or other active ingredient(s) of present invention along with protein or peptide antigens. The protein and/or peptide antigen will deliver a stimulatory signal to both B and T lymphocytes. B lymphocytes will respond to antigen through their surface immunoglobulin receptor. T lymphocytes will respond to antigen through the T cell receptor (TCR) following presentation of the antigen by MHC proteins. MHC.and structurally related proteins including those encoded by class I and class II MHC genes on host cells will serve to present the peptide antigen(s) to T lymphocytes. The antigen components could also be supplied as purified MHC-peptide complexes alone or with co-stimulatory molecules that can directly signal T cells. Alternatively antibodies able to bind surface immunoglobulin and other molecules on B cells as well as antibodies able to bind the TCR and other molecules on T cells can be combined with the pharmaceutical composition of the invention

The pharmaceutical composition of the invention may be in the form of a liposome in which protein of the present invention is combined, in addition to other pharmaceutically acceptable carriers, with amphipathic agents such as lipids which exist in aggregated form as micelles, insoluble monolayers, liquid crystals, or lamellar layers in aqueous solution. Suitable lipids for liposomal formulation include, without limitation, monoglycerides, diglycerides, sulfatides, lysolecithins, phospholipids, saponin, bile acids, and the like. Preparation of such liposomal formulations is within the level of skill in the art, as disclosed, for example, in U.S. Pat. Nos. 4,235,871; 4,501,728; 4,837,028; and 4,737,323, all of which are incorporated herein by reference.

The amount of protein or other active ingredient of the present invention in the pharmaceutical composition of the present invention will depend upon the nature and severity of the condition being treated, and on the nature of prior treatments which the patient has undergone. Ultimately, the attending physician will decide the amount of protein or other active ingredient of the present invention with which to treat each individual patient. Initially, the attending physician will administer low doses of protein or other active ingredient of the present invention and observe the patient's response. Larger doses of protein or other active ingredient of the present invention may be administered until the optimal therapeutic effect is obtained for the patient, and at that point the dosage is not increased further. It is contemplated that the various pharmaceutical compositions used to practice the method of the present invention should contain about 0.01 μg to about 100 mg (preferably about 0.1 μg to about 10 mg, more preferably about 0.1 μg to about 1 mg) of protein or other active ingredient of the present invention per kg body weight. For compositions of the present invention which are useful for bone, cartilage, tendon or ligament regeneration, the therapeutic method includes administering the composition topically, systematically, or locally as an implant or device. When administered, the therapeutic composition for use in this invention is, of course, in a pyrogen-free, physiologically acceptable form. Further, the composition may desirably be encapsulated or injected in a viscous form for delivery to the site of bone, cartilage or tissue damage. Topical administration may be suitable for wound healing and tissue repair. Therapeutically useful agents other than a protein or other active ingredient of the invention which may also optionally be included in the composition as described above, may alternatively or additionally, be administered simultaneously or sequentially with the composition in the methods of the invention. Preferably for bone and/or cartilage formation, the composition would include a matrix capable of delivering the protein-containing or other active ingredient-containing composition to the site of bone and/or cartilage damage, providing a structure for the developing bone and cartilage and optimally capable of being resorbed into the body. Such matrices may be formed of materials presently in use for other implanted medical applications.

The choice of matrix material is based on biocompatibility, biodegradability, mechanical properties, cosmetic appearance and interface properties. The particular application of the compositions will define the appropriate formulation. Potential matrices for the compositions may be biodegradable and chemically defined calcium sulfate, tricalcium phosphate, hydroxyapatite, polylactic acid, polyglycolic acid and polyanhydrides. Other potential materials are biodegradable and biologically well-defined, such as bone or dermal collagen. Further matrices are comprised of pure proteins or extracellular matrix components. Other potential matrices are nonbiodegradable and chemically defined, such as sintered hydroxyapatite, bioglass, aluminates, or other ceramics. Matrices may be comprised of combinations of any of the above mentioned types of material, such as polylactic acid and hydroxyapatite or collagen and tricalcium phosphate. The bioceramics may be altered in composition, such as in calcium-aluminate-phosphate and processing to alter pore size, particle size, particle shape, and biodegradability. Presently preferred is a 50:50 (mole weight) copolymer of lactic acid and glycolic acid in the form of porous particles having diameters ranging from 150 to 800 microns. In some applications, it will be useful to utilize a sequestering agent, such as carboxymethyl cellulose or autologous blood clot, to prevent the protein compositions from disassociating from the matrix.

A preferred family of sequestering agents is cellulosic materials such as alkylcelluloses (including hydroxyalkylcelluloses), including methylcellulose, ethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, hydroxypropyl-methylcellulose, and carboxymethylcellulose, the most preferred being cationic salts of carboxymethylcellulose (CMC). Other preferred sequestering agents include hyaluronic acid, sodium alginate, poly(ethylene glycol), polyoxyethylene oxide, carboxyvinyl polymer and poly(vinyl alcohol). The amount of sequestering agent useful herein is 0.5-20 wt %, preferably 1-10 wt % based on total formulation weight, which represents the amount necessary to prevent desorption of the protein from the polymer matrix and to provide appropriate handling of the composition, yet not so much that the progenitor cells are prevented from infiltrating the matrix, thereby providing the protein the opportunity to assist the osteogenic activity of the progenitor cells. In further compositions, proteins or other active ingredients of the invention may be combined with other agents beneficial to the treatment of the bone and/or cartilage defect, wound, or tissue in question. These agents include various growth factors such as epidermal growth factor (EGF), platelet derived growth factor (PDGF), transforming growth factors (TGF-α and TGF-β), and insulin-like growth factor (IGF).

The therapeutic compositions are also presently valuable for veterinary applications. Particularly domestic animals and thoroughbred horses, in addition to humans, are desired patients for such treatment with proteins or other active ingredients of the present invention. The dosage regimen of a protein-containing pharmaceutical composition to be used in tissue regeneration will be determined by the attending physician considering various factors which modify the action of the proteins, e.g., amount of tissue weight desired to be formed, the site of damage, the condition of the damaged tissue, the size of a wound, type of damaged tissue (e.g., bone), the patient's age, sex, and diet, the severity of any infection, time of administration and other clinical factors. The dosage may vary with the type of matrix used in the reconstitution and with inclusion of other proteins in the pharmaceutical composition. For example, the addition of other known growth factors, such as IGF I (insulin like growth factor I), to the final composition, may also effect the dosage. Progress can be monitored by periodic assessment of tissue/bone growth and/or repair, for example, X-rays, histomorphometric determinations and tetracycline labeling.

Polynucleotides of the present invention can also be used for gene therapy. Such polynucleotides can be introduced either in vivo or ex vivo into cells for expression in a mammalian subject. Polynucleotides of the invention may also be administered by other known methods for introduction of nucleic acid into a cell or organism (including, without limitation, in the form of viral vectors or naked DNA). Cells may also be cultured ex vivo in the presence of proteins of the present invention in order to proliferate or to produce a desired effect on or activity in such cells. Treated cells can then be introduced in vivo for therapeutic purposes.

3.12.3 Effective Dosage

Pharmaceutical compositions suitable for use in the present invention include compositions wherein the active ingredients are contained in an effective amount to achieve its intended purpose. More specifically, a therapeutically effective amount means an amount effective to prevent development of or to alleviate the existing symptoms of the subject being treated. Determination of the effective amount is well within the capability of those skilled in the art, especially in light of the detailed disclosure provided herein. For any compound used in the method of the invention, the therapeutically effective dose can be estimated initially from appropriate in vitro assays. For example, a dose can be formulated in animal models to achieve a circulating concentration range that can be used to more accurately determine useful doses in humans. For example, a dose can be formulated in animal models to achieve a circulating concentration range that includes the IC 50 as determined in cell culture (i.e., the concentration of the test compound which achieves a half-maximal inhibition of the protein's biological activity). Such information can be used to more accurately determine useful doses in humans.

A therapeutically effective dose refers to that amount of the compound that results in amelioration of symptoms or a prolongation of survival in a patient. Toxicity and therapeutic efficacy of such compounds can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, e.g., for determining the LD 50 (the dose lethal to 50% of the population) and the ED 50 (the dose therapeutically effective in 50% of the population). The dose ratio between toxic and therapeutic effects is the therapeutic index and it can be expressed as the ratio between LD 50 and ED 50 . Compounds which exhibit high therapeutic indices are preferred. The data obtained from these cell culture assays and animal studiescan be used in formulating a range of dosage for use in human. The dosage of such compounds lies preferably within a range of circulating concentrations that include the ED 50 with little or no toxicity. The dosage may vary within this range depending upon the dosage form employed and the route of administration utilized. The exact formulation, route of administration and dosage can be chosen by the individual physician in view of the patient's condition. See, e.g., Fingl et al., 1975, in “The Pharmacological Basis of Therapeutics”, Ch. 1 p.1. Dosage amount and interval may be adjusted individually to provide plasma levels of the active moiety which are sufficient to maintain the desired effects, or minimal effective concentration (MEC). The MEC will vary for each compound but can be estimated from in vitro data Dosages necessary to achieve the MEC will depend on individual characteristics and route of administration. However, HPLC assays or bioassays can be used to determine plasma concentrations.

Dosage intervals can also be determined using MEC value. Compounds should be administered using a regimen which maintains plasma levels above the MEC for 10-90% of the time, preferably between 30-90% and most preferably between 50-90%. In cases of local administration or selective uptake, the effective local concentration of the drug may not be related to plasma concentration.

An exemplary dosage regimen for polypeptides or other compositions of the invention will be in the range of about 0.01 μg/kg to 100 mg/kg of body weight daily, with the preferred dose being about 0.1 μg/kg to 25 mg/kg of patient body weight daily, varying in adults and children. Dosing may be once daily, or equivalent doses may be delivered at longer or shorter intervals.

The amount of composition administered will, of course, be dependent on the subject being treated, on the subject's age and weight, the severity of the affliction, the manner of administration and the judgment of the prescribing physician.

3.12.4 Packaging

The compositions may, if desired, be presented in a pack or dispenser device which may contain one or more unit dosage forms containing the active ingredient. The pack may, for example, comprise metal or plastic foil, such as a blister pack. The pack or dispenser device may be accompanied by instructions for administration. Compositions comprising a compound of the invention formulated in a compatible pharmaceutical carrier may also be prepared, placed in an appropriate container, and labeled for treatment of an indicated condition.

3.13 Antibodies

Also included in the invention are antibodies to proteins, or fragments of proteins of the invention. The term “antibody” as used herein refers to immunoglobulin molecules and immunologically active portions of immunoglobulin (Ig) molecules, i.e., molecules that contain an antigen-binding site that specifically binds (immunoreacts with) an antigen. Such antibodies include, but are not limited to, polyclonal, monoclonal, chimeric, single chain, F ab , F ab′ and F (ab′)2 fragments, and an F ab expression library. In general, an antibody molecule obtained from humans relates to any of the classes IgG, IgM, IgA, IgE and IgD, which differ from one another by the nature of the heavy chain present in the molecule. Certain classes have subclasses as well, such as IgG 1 , IgG 2 , and others. Furthermore, in humans, the light chain may be a kappa chain or a lambda chain. Reference herein to antibodies includes a reference to all such classes, subclasses and types of human antibody species.

An isolated related protein of the invention may be intended to serve as an antigen, or a portion or fragment thereof, and additionally can be used as an immunogen to generate antibodies that inmmunospecifically bind the antigen, using standard techniques for polyclonal and monoclonal antibody preparation. The full-length protein can be used or, alternatively, the invention provides antigenic peptide fragments of the antigen for use as immunogens. An antigenic peptide fragment comprises at least 6 mino acid residues of the amino acid sequence of the full length protein, such as an amino acid sequence shown in SEQ ID NO: 1-948, and encompasses an epitope thereof such that an antibody raised against the peptide forms a specific immune complex with the full length protein or with any fragment that contains the epitope. Preferably, the antigenic peptide comprises at least 10 amino acid residues, or at least 15 amino acid residues, or at least 20 amino acid residues, or at least 30 amino acid residues. Preferred epitopes encompassed by the antigenic peptide are regions of the protein that are located on its surface; commonly these are hydrophilic regions.

In certain embodiments of the invention, at least one epitope encompassed by the antigenic peptide is a region of alpha-2-macroglobulin-like protein that is located on the surface of the protein, e.g., a hydrophilic region. A hydrophobicity analysis of the human related protein sequence will indicate which regions of a related protein are particularly hydrophilic and, therefore, are likely to encode surface residues useful for targeting antibody production. As a means for targeting antibody production, hydropathy plots showing regions of hydrophilicity and hydrophobicity may be generated by any method well known in the art, including, for example, the Kyte Doolittle or the Hopp Woods methods, either with or without Fourier transformation. See, e.g., Hopp and Woods, 1981 , Proc. Nat. Acad. Sci. USA 78: 3824-3828; Kyte and Doolittle 1982 , J. Mol. Biol . 157: 105-142, each of which is incorporated herein by reference in its entirety. Antibodies that are specific for one or more domains within an antigenic protein, or derivatives, fragments, analogs or homologs thereof, are also provided herein.

A protein of the invention, or a derivative, fragment, analog, homolog or ortholog thereof, may be utilized as an immunogen in the generation of antibodies that immunospecifically bind these protein components.

The term “specific for” indicates that the variable regions of the antibodies of the invention recognize and bind polypeptides of the invention exclusively (i.e., able to distinguish the polypeptide of the invention from other similar polypeptides despite sequence identity, homology, or similarity found in the family of polypeptides), but may also interact with other proteins (for example, S. aureus protein A or other antibodies in ELISA techniques) through interactions with sequences outside the variable region of the antibodies, and in particular, in the constant region of the molecule. Screening assays to determine binding specificity of an antibody of the invention are well known and routinely practiced in the art. For a comprehensive discussion of such assays, see Harlow et al. (Eds), Antibodies A Laboratory Manual; Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y. (1988), Chapter 6. Antibodies that recognize and bind fragments of the polypeptides of the invention are also contemplated, provided that the antibodies are first and foremost specific for, as defined above, full-length polypeptides of the invention. As with antibodies that are specific for full length polypeptides of the invention, antibodies of the invention that recognize fragments are those which can distinguish polypeptides from the same family of polypeptides despite inherent sequence identity, homology, or similarity found in the family of proteins.

Antibodies of the invention are useful for, for example, therapeutic purposes (by modulating activity of a polypeptide of the invention), diagnostic purposes to detect or quantitate a polypeptide of the invention, as well as purification of a polypeptide of the invention. Kits comprising an antibody of the invention for any of the purposes described herein are also comprehended. In general, a kit of the invention also includes a control antigen for which the antibody is immunospecific. The invention further provides a hybridoma that produces an antibody according to the invention. Antibodies of the invention are useful for detection and/or purification of the polypeptides of the invention.

Monoclonal antibodies binding to the protein of the invention may be useful diagnostic agents for the immunodetection of the protein. Neutralizing monoclonal antibodies binding to the protein may also be useful therapeutics for both conditions associated with the protein and also in the treatment of some forms of cancer where abnormal expression of the protein is involved. In the case of cancerous cells or leukemic cells, neutralizing monoclonal antibodies against the protein may be useful in detecting and preventing the metastatic spread of the cancerous cells, which may be mediated by the protein.

The labeled antibodies of the present invention can be used for in vitro, in vivo, and in situ assays to identify cells or tissues in which a fragment of the polypeptide of interest is expressed. The antibodies may also be used directly in therapies or other diagnostics. The present invention further provides the above-described antibodies immobilized on a solid support. Examples of such solid supports include plastics such as polycarbonate, complex carbohydrates such as agarose and Sepharose®, acrylic resins and such as polyacrylamide and latex beads. Techniques for coupling antibodies to such solid supports are well known in the art (Weir, D. M. et al., “Handbook of Experimental Immunology” 4th Ed., Blackwell Scientific Publications, Oxford, England, Chapter 10 (1986); Jacoby, W. D. et al., Meth. Enzym. 34 Academic Press, N.Y. (1974)). The immobilized antibodies of the present invention can be used for in vitro, in vivo, and in situ assays as well as for immuno-affinity purification of the proteins of the present invention.

Various procedures known within the art may be used for the production of polyclonal or monoclonal antibodies directed against a protein of the invention, or against derivatives, fragments, analogs homologs or orthologs thereof (see, for example, Antibodies: A Laboratory Manual, Harlow E, and Lane D, 1988, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., incorporated herein by reference). Some of these antibodies are discussed below.

3.13.1 Polyclonal Antibodies

For the production of polyclonal antibodies, various suitable host animals (e.g., rabbit, goat, mouse or other mammal) may be immunized by one or more injections with the native protein, a synthetic variant thereof, or a derivative of the foregoing. An appropriate immunogenic preparation can contain, for example, the naturally occurring immunogenic protein, a chemically synthesized polypeptide representing the immunogenic protein, or a recombinantly expressed immunogenic protein. Furthermore, the protein may be conjugated to a second protein known to be immunogenic in the mammal being immunized. Examples of such immunogenic proteins include but are not limited to keyhole limpet hemocyanin, serum albumin, bovine thyroglobulin, and soybean trypsin inhibitor. The preparation can further include an adjuvant. Various adjuvants used to increase the immunological response include, but are not limited to, Freund's (complete and incomplete), mineral gels (e.g., aluminum hydroxide), surface-active substances (e.g., lysolecithin, pluronic polyols, polyanions, peptides, oil emulsions, dinitrophenol, etc.), adjuvants usable in humans such as Bacille Calmette-Guerin and Corynebacterium parvum, or similar immunostimulatory agents. Additional examples of adjuvants that can be employed include MPL-TDM adjuvant (monophosphoryl Lipid A, synthetic trehalose dicorynomycolate).

The polyclonal antibody molecules directed against the immunogenic protein can be isolated from the manmmal (e.g., from the blood) and further purified by well known techniques, such as affinity chromatography using protein A or protein G, which provide primarily the IgG faction of immune serum. Subsequently, or alternatively, the specific antigen which is the target of the immunoglobulin sought, or an epitope thereof, may be immobilized on a column to purify the immune specific antibody by immunoaffinity chromatography. Purification of inmmunoglobulins is discussed, for example, by D. Wilkinson (The Scientist, published by The Scientist, Inc., Philadelphia Pa., Vol. 14, No. 8 (Apr. 17, 2000), pp. 25-28).

3.13.2 Monoclonal Antibodies

The term “monoclonal antibody” (MAb) or “monoclonal antibody composition”, as used herein, refers to a population of antibody molecules that contain only one molecular species of antibody molecule consisting of a unique light chain gene product and a unique heavy chain gene product. In particular, the complementarity determining regions (CDRs) of the monoclonal antibody are identical in all the molecules of the population. MAbs thus contain an antigen-binding site capable of immunoreacting with a particular epitope of the antigen characterized by a unique binding affinity for it.

Monoclonal antibodies can be prepared using hybridoma methods, such as those described by Kohler and Milstein, Nature , 256:495 (1975). In a hybridoma method, a mouse, hamster, or other appropriate host animal, is typically immunized with an immunizing agent to elicit lymphocytes that produce or are capable of producing antibodies that will specifically bind to the immunizing agent. Alternatively, the lymphocytes can be immunized in vitro.

The immunizing agent will typically include the protein antigen, a fragment thereof or a fusion protein thereof. Generally, either peripheral blood lymphocytes are used if cells of human origin are desired, or spleen cells or lymph node cells are used if non-human mammalian sources are desired. The lymphocytes are then fused with an immortalized cell line using a suitable fusing agent, such as polyethylene glycol, to form a hybridoma cell (Goding, Monoclonal Antibodies: Principles and Practice , Academic Press, (1986) pp. 59-103). Immortalized cell lines are usually transformed mammalian cells, particularly myeloma cells of rodent, bovine and human origin. Usually, rat or mouse myeloma cell lines are employed. The hybridoma cells can be cultured in a suitable culture medium that preferably contains one or more substances that inhibit the growth or survival of the unfused, immortalized cells. For example, if the parental cells lack the enzyme hypoxanthine guanine phosphoribosyl transferase (HGPRT or HPRT), the culture medium for the hybridomas typically will include hypoxanthine, aminopterin, and thymidine (“HAT medium”), which substances prevent the growth of HGPRT-deficient cells.

Preferred immortalized cell lines are those that fuse efficiently, support stable high level expression of antibody by the selected antibody-producing cells, and are sensitive to a medium such as HAT medium. More preferred immortalized cell lines are murine myeloma lines, which can be obtained, for instance, from the Salk Institute Cell Distribution Center, San Diego, Calif. and the American Type Culture Collection, Manassas, Va. Human myeloma and mouse-human heteromyeloma cell lines also have been described for the production of human monoclonal antibodies (Kozbor, J. Immunol ., 133:3001 (1984); Brodeur et al., Monoclonal Antibody Production Techniques and Applications , Marcel Dekker, Inc., New York, (1987) pp. 51-63).

The culture medium in which the hybridoma cells are cultured can then be assayed for the presence of monoclonal antibodies directed against the antigen. Preferably, the binding specificity of monoclonal antibodies produced by the hybridoma cells is determined by immunoprecipitation or by an in vitro binding assay, such as radioimmunoassay (RIA) or enzyme-linked immunoabsorbent assay (ELISA). Such techniques and assays are known in the art. The binding affinity of the monoclonal antibody can, for example, be determined by the Scatchard analysis of Munson and Pollard, Anal. Biochem ., 107:220 (1980). Preferably, antibodies having a high degree of specificity and a high binding affinity for the target antigen are isolated.

After the desired hybridoma cells are identified, the clones can be subcloned by limiting dilution procedures and grown by standard methods. Suitable culture media for this purpose include, for example, Dulbecco's Modified Eagle's Medium and RPMI-1640 medium. Alternatively, the hybridoma cells can be grown in vivo as ascites in a mammal.

The monoclonal antibodies secreted by the subclones can be isolated or purified from the culture medium or ascites fluid by conventional immunoglobulin purification procedures such as, for example, protein A-Sepharose, hydroxylapatite chromatography, gel electrophoresis, dialysis, or affinity chromatography.

The monoclonal antibodies can also be made by recombinant DNA methods, such as those described in U.S. Pat. No. 4,816,567. DNA encoding the monoclonal antibodies of the invention can be readily isolated and sequenced using conventional procedures (e.g., by using oligonucleotide probes that are capable of binding specifically to genes encoding the heavy and light chains of murine antibodies). The hybridoma cells of the invention serve as a preferred source of such DNA. Once isolated, the DNA can be placed into expression vectors, which are then transfected into host cells such as simian COS cells, Chinese hamster ovary (CHO) cells, or myeloma cells that do not otherwise produce immunoglobulin protein, to obtain the synthesis of monoclonal antibodies in the recombinant host cells. The DNA also can be modified, for example, by substituting the coding sequence for human heavy and light chain constant domains in place of the homologous murine sequences (U.S. Pat. No. 4,816,567; Morrison, Nature 368, 812-13 (1994)) or by covalently joining to the immunoglobulin coding sequence all or part of the coding sequence for a non-immunoglobulin polypeptide. Such a non-immunoglobulin polypeptide can be substituted for the constant domains of an antibody of the invention, or can be substituted for the variable domains of one antigen-combining site of an antibody of the invention to create a chimeric bivalent antibody.

3.13.3 Humanized Antibodies

The antibodies directed against the protein antigens of the invention can further comprise humanized antibodies or human antibodies. These antibodies are suitable for administration to humans without engendering an immune response by the human against the administered immunoglobulin. Humanized forms of antibodies are chimeric immunoglobulins, immunoglobulin chains or fragments thereof (such as Fv, Fab, Fab′, F(ab′) 2 or other antigen-binding subsequences of antibodies) that are principally comprised of the sequence of a human immunoglobulin, and contain minimal sequence derived from a non-human immunoglobulin. Humanization can be performed following the method of Winter and co-workers (Jones et al., Nature , 321:522-525 (1986); Riechmann et al., Nature , 332:323-327 (1988); Verhoeyen et al., Science , 239:1534-1536 (1988)), by substituting rodent CDRs or CDR sequences for the corresponding sequences of a human antibody. (See also U.S. Pat. No. 5,225,539). In some instances, Fv framework residues of the human immunoglobulin are replaced by corresponding non-human residues. Humanized antibodies can also comprise residues that are found neither in the recipient antibody nor in the imported CDR or framework sequences. In general, the humanized antibody will comprise substantially all of at least one, and typically two, variable domains, in which all or substantially all of the CDR regions correspond to those of a non-human immunoglobulin and all or substantially all of the framework regions are those of a human immunoglobulin consensus sequence. The humanized antibody optimally also will comprise at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin (Jones et al., 1986; Riechmann et al., 1988; and Presta, Curr. Op. Struct. Biol ., 2:593-596 (1992)).

3.13.4 Human Antibodies

Fully human antibodies relate to antibody molecules in which essentially the entire sequences of both the light chain and the heavy chain, including the CDRs, arise from human genes. Such antibodies are termed “human antibodies”, or “fully human antibodies” herein. Human monoclonal antibodies can be prepared by the trioma technique; the human B-cell hybridoma technique (see Kozbor, et al., 1983 Immunol Today 4: 72) and the EBV hybridoma technique to produce human monoclonal antibodies (see Cole, et al., 1985 In: M ONOCLONAL A NTIBODIES AND C ANCER T HERAPY , Alan R. Liss, Inc., pp. 77-96). Human monoclonal antibodies may be utilized in the practice of the present invention and may be produced by using human hybridomas (see Cote, et al., 1983. Proc Natl Acad Sci USA 80: 2026-2030) or by transforming human B-cells with Epstein Barr Virus in vitro (see Cole, et al., 1985 In: M ONOCLONAL A NTIBODIES AND C ANCER T HERAPY , Alan R. Liss, Inc., pp. 77-96).

In addition, human antibodies can also be produced using additional techniques, including phage display libraries (Hoogenboom and Winter, J. Mol. Biol ., 227:381 (1991); Marks et al., J. Mol. Biol ., 222:581 (1991)). Similarly, human antibodies can be made by introducing human immunoglobulin loci into transgenic animals, e.g., mice in which the endogenous immunoglobulin genes have been partially or completely inactivated. Upon challenge, human antibody production is observed, which closely resembles that seen in humans in all respects, including gene rearrangement, assembly, and antibody repertoire. This approach is described, for example, in U.S. Pat. Nos. 5,545,807; 5,545,806; 5,569,825; 5,625,126; 5,633,425; 5,661,016, and in Marks et al. ( Bio/Technology 10, 779-783 (1992)); Lonberg et al. ( Nature 368 856-859 (1994)); Morrison (Nature 368, 812-13 (1994)); Fishwild et al, ( Nature Biotechnology 14, 845-51 (1996)); Neuberger ( Nature Biotechnology 14, 826 (1996)); and Lonberg and Huszar ( Intern. Rev. Immunol . 13 65-93 (1995)).

Human antibodies may additionally be produced using taansgenic nonhuman animals that are modified so as to produce fully human antibodies rather than the animal's endogenous antibodies in response to challenge by an antigen. (See PCT publication WO94/02602). The endogenous genes encoding the heavy and light immunoglobulin chains in the nonhuman host have been incapacitated, and active loci encoding human heavy and light chain immunoglobulins are inserted into the host's genome. The human genes are incorporated, for example, using yeast artificial chromosomes containing the requisite human DNA segments. An animal which provides all the desired modifications is then obtained as progeny by crossbreeding intermediate transgenic animals containing fewer than the full complement of the modifications. The preferred embodiment of such a nonhuman animal is a mouse, and is termed the Xenomouse™ as disclosed in PCT publications WO 96/33735 and WO 96/34096. This animal produces B cells that secrete fully human immunoglobulins. The antibodies can be obtained directly from the animal after immunization with an immunogen of interest, as, for example, a preparation of a polyclonal antibody, or alternatively from immortalized B cells derived from the animal, such as hybridomas producing monoclonal antibodies. Additionally, the genes encoding the immunoglobulins with human variable regions can be recovered and expressed to obtain the antibodies directly, or can be further modified to obtain analogs of antibodies such as, for example, single chain Fv molecules.

An example of a method of producing a nonhuman host, exemplified as a mouse, lacking expression of an endogenous immunoglobulin heavy chain is disclosed in U.S. Pat. No. 5,939,598. It can be obtained by a method including deleting the J segment genes from at least one endogenous heavy chain locus in an embryonic stem cell to prevent rearrangement of the locus and to prevent formation of a transcript of a rearranged immunoglobulin heavy chain locus, the deletion being effected by a targeting vector containing a gene encoding a selectable marker; and producing from the embryonic stem cell a transgenic mouse whose somatic and germ cells contain the gene encoding the selectable marker.

A method for producing an antibody of interest, such as a human antibody, is disclosed in U.S. Pat. No. 5,916,771. It includes introducing an expression vector that contains a nucleotide sequence encoding a heavy chain into one mammalian host cell in culture, introducing an expression vector containing a nucleotide sequence encoding a light chain into another mammalian host cell, and fusing the two cells to form a hybrid cell. The hybrid cell expresses an antibody containing the heavy chain and the light chain.

In a further improvement on this procedure, a method for identifying a clinically relevant epitope on an imnmunogen, and a correlative method for selecting an antibody that binds immunospecifically to the relevant epitope with high affinity, are disclosed in PCT publication WO 99/53049.

3.13.5 Fab Fragments and Single Chain Antibodies

According to the invention, techniques can be adapted for the production of single-chain antibodies specific to an antigenic protein of the invention (see e.g., U.S. Pat. No. 4,946,778). In addition, methods can be adapted for the construction of F ab expression libraries (see e.g., Huse, et al., 1989 Science 246: 1275-1281) to allow rapid and effective identification of monoclonal F ab fragments with the desired specificity for a protein or derivatives, fragments, analogs or homologs thereof. Antibody fragments that contain the idiotypes to a protein antigen may be produced by techniques known in the art including, but not limited to: (i) an F (ab′)2 fragment produced by pepsin digestion of an antibody molecule; (ii) an F ab fragment generated by reducing the disulfide bridges of an F (ab′)2 fragment; (iii) an F ab fragment generated by the treatment of the antibody molecule with papain and a reducing agent and (iv) F v fragments.

3.13.6 Bispecific Antibodies

Bispecific antibodies are monoclonal, preferably human or humanized, antibodies that have binding specificities for at least two different antigens. In the present case, one of the binding specificities is for an antigenic protein of the invention. The second binding target is any other antigen, and advantageously is a cell-surface protein or receptor or receptor subunit.

Methods for making bispecific antibodies are known in the art. Traditionally, the recombinant production of bispecific antibodies is based on the co-exprcssion of two immunoglobulin heavyhain/light-chain pairs, where the two heavy chains have different specificities (Milstein and Cuello, Nature , 305:537-539 (1983)). Because of the random assortment of immunoglobulin heavy and light chains, these hybridomas (quadromas) produce a potential mixture of ten different antibody molecules, of which only one has the correct bispecific structure. The purification of the correct molecule is usually accomplished by affinity chromatography steps. Similar procedures are disclosed in WO 93/08829, published 13 May 1993, and in Traunecker et al., 1991 EMBO J ., 10:3655-3659.

Antibody variable domains with the desired binding specificities (antibody-antigen combining sites) can be fused to immunoglobulin constant domain sequences. The fusion preferably is with an immunoglobulin heavy-chain constant domain, comprising at least part of the hinge, CH2, and CH3 regions. It is preferred to have the first heavy-chain constant region (CH1) containing the site necessary for light-chain binding present in at least one of the fusions. DNAs encoding the immunoglobulin heavy-chain fusions and, if desired, the immunoglobulin light chain, are inserted into separate expression vectors, and are co-transfected into a suitable host organism. For further details of generating bispecific antibodies see, for example, Suresh et al., Methods in Enzymology , 121:210 (1986).

According to another approach described in WO 96/27011, the interface between a pair of antibody molecules can be engineered to maximize the percentage of heterodimers that are recovered from recombinant cell culture. The preferred interface comprises at least a part of the CH3 region of an antibody constant domain. In this method, one or more small amino acid side chains from the interface of the first antibody molecule are replaced with larger side chains (e.g. tyrosine or tryptophan). Compensatory “cavities” of identical or similar size to the large side chain(s) are created on the interface of the second antibody molecule by replacing large amino acid side chains with smaller ones (e.g. alanine or threonine). This provides a mechanism for increasing the yield of the heterodimer over other unwanted end-products such as homodimers.

Bispecific antibodies can be prepared as full length antibodies or antibody fragments (e.g. F(ab′) 2 bispecific antibodies). Techniques for generating bispecific antibodies from antibody fragments have been described in the literature. For example, bispecific antibodies can be prepared using chemical linkage. Brennan et al., Science 229:81 (1985) describe a procedure wherein intact antibodies are proteolytically cleaved to generate F(ab′) 2 fragments. These fragments are reduced in the presence of the dithiol complexing agent sodium arsenite to stabilize vicinal dithiols and prevent intermolecular disulfide formation. The Fab′ fragments generated are then converted to thionitrobenzoate (TNB) derivatives. One of the Fab′-TNB derivatives is then reconverted to the Fab′-thiol by reduction with mercaptoethylamine and is mixed with an equimolar amount of the other Fab′-TNB derivative to form the bispecific antibody. The bispecific antibodies produced can be used as agents for the selective immobilization of enzymes.

Additionally, Fab′ fragments can be directly recovered from E. coli and chemically coupled to form bispecific antibodies. Shalaby et al., J. Exp. Med . 175:217-225 (1992) describe the production of a fully humanized bispecific antibody F(ab′) 2 molecule. Each Fab′ fragment was separately secreted from E. coli and subjected to directed chemical coupling in vitro to form the bispecific antibody. The bispecific antibody thus formed was able to bind to cells overexpressing the ErbB2 receptor and normal human T cells, as well as trigger the lytic activity of human cytotoxic lymphocytes against human breast tumor targets.

Various techniques for making and isolating bispecific antibody fragments directly from recombinant cell culture have also been described. For example, bispecific antibodies have been produced using leucine zippers. Kostelny et al., J. Immunol . 148(5):1547-1553 (1992). The leucine zipper peptides from the Fos and Jun proteins were linked to the Fab′ portions of two different antibodies by gene fusion. The antibody homodimers were reduced at the hinge region to form monomers and then re-oxidized to form the antibody heterodimers. This method can also be utilized for the production of antibody homodimers. The “diabody” technology described by Hollinger et al., Proc. Natl. Acad. Sci. USA 90:6444-6448 (1993) has provided an alternative mechanism for making bispecific antibody fragments. The fragments comprise a heavy-chain variable domain (V H ) connected to a lighthain variable domain (V L ) by a linker which is too short to allow pairing between the two domains on the same chain. Accordingly, the V H and V L domains of one fragment are forced to pair with the complementary V L and V H domains of another fragment, thereby forming two antigen-binding sites. Another strategy for making bispecific antibody fragments by the use of single-chain Fv (sFv) dimers has also been reported. See, Gruber et al., J. Immunol . 152:5368 (1994).

Antibodies with more than two valencies are contemplated. For example, trispecific antibodies can be prepared. Tutt et al., J. Immunol . 147:60 (1991).

Exemplary bispecific antibodies can bind to two different epitopes, at least one of which originates in the protein antigen of the invention. Alternatively, an anti-antigenic arm of an immunoglobulin molecule can be combined with an arm which binds to a triggering molecule on a leukocyte such as a T-cell receptor molecule (e.g. CD2, CD3, CD28, or B7), or Fc receptors for IgG (Fc R), such as Fc RI (CD64), Fe RII (CD32) and Fc RIII (CD16) so as to focus cellular defense mechanisms to the cell expressing the particular antigen. Bispecific antibodies can also be used to direct cytotoxic agents to cells which express a particular antigen. These antibodies possess an antigen-binding arm and an arm which binds a cytotoxic agent or a radionuclide chelator, such as EOTUBE, DPTA, DOTA, or TETA. Another bispecific antibody of interest binds the protein antigen described herein and further binds tissue factor TF).

3.13.7 Heteroconjugate Antibodies

Heteroconjugate antibodies are also within the scope of the present invention. Heteroconjugate antibodies are composed of two covalently joined antibodies. Such antibodies have, for example, been proposed to target immune system cells to unwanted cells (U.S. Pat. No. 4,676,980), and for treatment of HIV infection (WO 91/00360; WO 92/200373; EP 03089). It is contemplated that the antibodies can be prepared in vitro using known methods in synthetic protein chemistry, including those involving crosslinking agents. For example, immunotoxins can be constructed using a disulfide exchange reaction or by forming a thioether bond. Examples of suitable reagents for this purpose include iminothiolate and methyl-4-mercaptobutyrimidate and those disclosed, for example, in U.S. Pat. No. 4,676,980.

3.13.8 Effector Function Engineering

It can be desirable to modify the antibody of the invention with respect to effector function, so as to enhance, e.g., the effectiveness of the antibody in treating cancer. For example, cysteine residue(s) can be introduced into the Fe region, thereby allowing interchain disulfide bond formation in this region. The homodimeric antibody thus generated can have improved internalization capability and/or increased complement-mediated cell killing and antibody-dependent cellular cytotoxicity (ADCC). See Caron et al., J. Exp Med., 176: 1191-1195 (1992) and Shopes, J. Imunol., 148: 2918-2922 (1992). Homodimeric antibodies with enhanced anti-tumor activity can also be prepared using heterobifunctional cross-linkers as described in Wolff et al. Cancer Research, 53: 2560-2565 (1993). Alternatively, an antibody can be engineered that has dual Fc regions and can thereby have enhanced complement lysis and ADCC capabilities. See Stevenson et al., Anti-Cancer Drug Design, 3: 219-230 (1989).

3.13.9 Immunoconjugates

The invention also pertains to immunoconjugates comprising an antibody conjugated to a cytotoxic agent such as a chemotherapeutic agent, toxin (e.g., an enzymatically active toxin of bacterial, fungal, plant, or animal origin, or fragments thereof), or a radioactive isotope (i.e., a radioconjugate).

Chemotherapeutic agents useful in the generation of such immunoconjugates have been described above. Enzymatically active toxins and fragments thereof that can be used include diphtheria A chain, nonbinding active fragments of diphtheria toxin, exotoxin A chain (from Pseudomonas aeruginosa), ricin A chain, abrin A chain, modeccin A chain, alpha-sarcin, Aleurites fordii proteins, dianthin proteins, Phytolaca americana proteins (PAPI, PAPII, and PAP-S), momordica charantia inhibitor, curcin, crotin, sapaonaria officinalis inhibitor, gelonin, mitogellin, restrictocin, phenomycin, enomycin, and the tricothecenes. A variety of radionuclides are available for the production of radioconjugated antibodies. Examples include 212 Bi, 131 I, 131 In, 90 y, and 186 Re.

Conjugates of the antibody and cytotoxic agent are made using a variety of bifunctional protein-coupling agents such as N-succinimidyl-3-(2-pyridyldithiol) propionate (SPDP), iminothiolane (IT), bifunctional derivatives of imidoesters (such as dimethyl adipimidate HCL), active esters (such as disuccinimidyl suberate), aldehydes (such as glutareldehyde), bis-azido compounds (such as bis (pazidobenzoyl) hexanediamine), bis-diazonium derivatives (such as bis-(p-diazoniumbenzoyl)-ethylenediamine), diisocyanates (such as tolyene 2,6-diisocyanate), and bis-active fluorine compounds (such as 1,5-difluoro-2,4-dinitrobenzene). For example, a ricin immunotoxin can be prepared as described in Vitetta et al., Science, 238: 1098 (1987). Carbon-14-labeled 1-isothiocyanatobenzyl-3-methyldiethylene triaminepentaacetic acid (MX-DTPA) is an exemplary chelating agent for conjugation of radionucleotide to the antibody. See WO94/11026.

In another embodiment, the antibody can be conjugated to a “receptor” (such streptavidin) for utilization in tumor pretargeting wherein the antibody-receptor conjugate is administered to the patient, followed by removal of unbound conjugate from the circulation using a clearing agent and then administration of a “ligand” (e.g., avidin) that is in turn conjugated to a cytotoxic agent.

3.14 Computer Readable Sequences

In one application of this embodiment, a nucleotide sequence of the present invention can be recorded on computer readable media. As used herein, “computer readable media” refers to any medium which can be read and accessed directly by a computer. Such media include, but are not limited to: magnetic storage media, such as floppy discs, hard disc storage medium, and magnetic tape; optical storage media such as CD-ROM; electrical storage media such as RAM and ROM; and hybrids of these categories such as magnetic/optical storage media. A skilled artisan can readily appreciate how any of the presently known computer readable mediums can be used to create a manufacture comprising computer readable medium having recorded thereon a nucleotide sequence of the present invention. As used herein, “recorded” refers to a process for storing information on computer readable medium. A skilled artisan can readily adopt any of the presently known methods for recording information on computer readable medium to generate manufactures comprising the nucleotide sequence information of the present invention.

A variety of data storage structures are available to a skilled artisan for creating a computer readable medium having recorded thereon a nucleotide sequence of the present invention. The choice of the data storage structure will generally be based on the means chosen to access the stored information. In addition, a variety of data processor programs and formats can be used to store the nucleotide sequence information of the present invention on computer readable medium. The sequence information can be represented in a word processing text file, formatted in commercially-available software such as WordPerfect and Microsoft Word, or represented in the form of an ASCII file, stored in a database application, such as DB2, Sybase, Oracle, or the like. A skilled artisan can readily adapt any number of data processor structuring formats (e.g. text file or database) in order to obtain computer readable medium having recorded thereon the nucleotide sequence information of the present invention.

By providing any of the nucleotide sequences SEQ ID NOs: 1-948 or a representative fragment thereof; or a nucleotide sequence at least 95% identical to any of the nucleotide sequences of SEQ ID NOs: 1-948 in computer readable form, a skilled artisan can routinely access the sequence information for a variety of purposes. Computer software is publicly available which allows a skilled artisan to access sequence information provided in a computer readable medium. The examples which follow demonstrate how software which implements the BLAST (Altschul et al., J. Mol. Biol. 215:403-410 (1990)) and BLAZE (Brutlag et al., Comp. Chem. 17:203-207 (1993)) search algorithms on a Sybase system is used to identify open reading frames (ORFs) within a nucleic acid sequence. Such ORFs may be protein encoding fragments and may be useful in producing commercially important proteins such as enzymes used in fermentation reactions and in the production of commercially useful metabolites.

As used herein, “a computer-based system” refers to the hardware means, software means, and data storage means used to analyze the nucleotide sequence information of the present invention. The minimum hardware means of the computer-based systems of the present invention comprises a central processing unit (CPU), input means, output means, and data storage means. A skilled artisan can readily appreciate that any one of the currently available computer-based systems are suitable for use in the present invention. As stated above, the computer-based systems of the present invention comprise a data storage means having stored therein a nucleotide sequence of the present invention and the necessary hardware means and software means for supporting and implementing a search means. As used herein, “data storage means” refers to memory which can store nucleotide sequence information of the present invention, or a memory access means which can access manufactures having recorded thereon the nucleotide sequence information of the present invention.

As used herein, “search means” refers to one or more programs which are implemented on the computer-based system to compare a target sequence or target structural motif with the sequence information stored within the data storage means. Scarch means are used to identify fragments or regions of a known sequence which match a particular target sequence or target motif A variety of known algorithms are disclosed publicly and a variety of commercially available software for conducting search means are and can be used in the computer-based systems of the present invention. Examples of such software includes, but is not limited to, Smnith-Waterman, MacPattern (EMBL), BLASTN and BLASTA (NPOLYPEPTIDEIA). A skilled artisan can readily recognize that any one of the available algorithms or implementing software packages for conducting homology searches can be adapted for use in the present computer-based systems. As used herein, a “target sequence” can be any nucleic acid or amino acid sequence of six or more nucleotides or two or more amino acids. A skilled artisan can readily recognize that the longer a target sequence is, the less likely a target sequence will be present as a random occurrence in the database. The most preferred sequence length of a target sequence is from about 10 to 300 amino acids, more preferably from about 30 to 100 nucleotide residues. However, it is well recognized that searches for commercially important fragments, such as sequence fragments involved in gene expression and protein processing, may be of shorter length.

As used herein, “a target structural motif,” or “target motif,” refers to any rationally selected sequence or combination of sequences in which the sequence(s) are chosen based on a three-dimensional configuration which is formed upon the folding of the target motif. There are a variety of target motifs known in the art. Protein target motifs include, but are not limited to, enzyme active sites and signal sequences. Nucleic acid target motifs include, but are not limited to, promoter sequences, hairpin structures and inducible expression elements (protein binding sequences).

3.15 Triple Helix Formation

In addition, the fragments of the present invention, as broadly described, can be used to control gene expression through triple helix formation or antisense DNA or RNA, both of which methods are based on the binding of a polynucleotide sequence to DNA or RNA. Polynucleotides suitable for use in these methods are preferably 20 to 40 bases in length and are designed to be complementary to a region of the gene involved in transcription (triple helix—see Lee et al., Nucl. Acids Res. 6:3073 (1979); Cooney et al., Science 15241:456 (1988); and Dervan et al., Science 251:1360 (1991)) or to the mRNA itself (antisense—Olmno, J. Neurochem. 56:560 (1991); Oligodeoxynucleotides as Antisense Inhibitors of Gene Expression, CRC Press, Boca Raton, Fla. (1988)). Triple helix-formation optimally results in a shut-off of RNA transcription from DNA, while antisense RNA hybridization blocks translation of an mRNA molecule into polypeptide. Both techniques have been demonstrated to be effective in model systems. Information contained in the sequences of the present invention is necessary for the design of an antisense or triple helix oligonucleotide.

3.16 Diagnostic Assays and Kits

The present invention further provides methods to identify the presence or expression of one of the ORFs of the present invention, or homolog thereof, in a test sample, using a nucleic acid probe or antibodies of the present invention, optionally conjugated or otherwise associated with a suitable label.

In general, methods for detecting a polynucleodide of the invention can comprise contacting a sample with a compound that binds to and forms a complex with the polynucleotide for a period sufficient to form the complex, and detecting the complex, so that if a complex is detected, a polynucleotide of the invention is detected in the sample. Such methods can also comprise contacting a sample under stringent hybridization conditions with nucleic acid primers that anneal to a polynucleotide of the invention under such conditions, and amplifying annealed polynucleotides, so that if a polynucleotide is amplified, a polynucleotide of the invention is detected in the sample.

In general, methods for detecting a polypeptide of the invention can comprise contacting a sample with a compound that binds to and forms a complex with the polypeptide for a period sufficient to form the complex, and detecting the complex, so that if a complex is detected, a polypeptide of the invention is detected in the sample.

In detail, such methods comprise incubating a test sample with one or more of the antibodies or one or more of the nucleic acid probes of the present invention and assaying for binding of the nucleic acid probes or antibodies to components within the test sample.

Conditions for incubating a nucleic acid probe or antibody with a test sample vary. Incubation conditions depend on the format employed in the assay, the detection methods employed, and the type and nature of the nucleic acid probe or antibody used in the assay. One skilled in the art will recognize that any one of the commonly available hybridization, amplification or immunological assay formats can readily be adapted to employ the nucleic acid probes or antibodies of the present invention. Examples of such assays can be found in Chard, T., An Introduction to Radioimmunoassay and Related Techniques, Elsevier Science Publishers, Amsterdam, The Netherlands (1986); Bullock, G. R. et al., Techniques in Immunocytochemistry, Academic Press, Orlando, Fla. Vol. 1 (1982), Vol. 2 (1983), Vol. 3 (1985); Tijssen, P., Practice and Theory of immunoassays: Laboratory Techniques in Biochemistry and Molecular Biology, Elsevier Science Publishers, Amsterdam, The Netherlands (1985). The test samples of the present invention include cells, protein or membrane extracts of cells, or biological fluids such as sputum, blood, serum, plasma, or urine. The test sample used in the above-described method will vary based on the assay format, nature of the detection method and the tissues, cells or extracts used as the sample to be assayed. Methods for preparing protein extracts or membrane extracts of cells are well known in the art and can be readily be adapted in order to obtain a sample which is compatible with the system utilized.

In another embodiment of the present invention, kits are provided which contain the necessary reagents to carry out the assays of the present invention. Specifically, the invention provides a compartment kit to receive, in close confinement, one or more containers which comprises: (a) a first container comprising one of the probes or antibodies of the present invention; and (b) one or more other containers comprising one or more of the following: wash reagents, reagents capable of detecting presence of a bound probe or antibody.

In detail, a compartment kit includes any kit in which reagents are contained in separate containers. Such containers include small glass containers, plastic containers or strips of plastic or paper. Such containers allows one to efficiently transfer reagents from one compartment to another compartment such that the samples and reagents are not cross-contaminated, and the agents or solutions of each container can be added in a quantitative fashion from one compartment to another. Such containers will include a container which will accept the test sample, a container which contains the antibodies used in the assay, containers which contain wash reagents (such as phosphate buffered saline, Tris-buffers, etc.), and containers which contain the reagents used to detect the bound antibody or probe. Types of detection reagents include labeled nucleic acid probes, labeled secondary antibodies, or in the alternative, if the primary antibody is labeled, the enzymatic, or antibody binding reagents which are capable of reacting with the labeled antibody. One skilled in the art will readily recognize that the disclosed probes and antibodies of the present invention can be readily incorporated into one of the established kit formats which are well known in the art.

3.17 Medical Imaging

The novel polypeptides and binding partners of the invention are useful in medical imaging of sites expressing the molecules of the invention (e.g., where the polypeptide of the invention is involved in the immune response, for imaging sites of inflammation or infection). See, e.g., Kunkel et al., U.S. Pat. No. 5,413,778. Such methods involve chemical attachment of a labeling or imaging agent, administration of the labeled polypeptide to a subject in a pharmaceutically acceptable carrier, and imaging the labeled polypeptide in vivo at the target site.

3.18 Screening Assays

Using the isolated proteins and polynucleotides of the invention, the present invention further provides methods of obtaining and identifying agents which bind to a polypeptide encoded by an ORF corresponding to any of the nucleotide sequences set forth in SEQ ID NOs: 1-948, or bind to a specific domain of the polypeptide encoded by the nucleic acid. In detail, said method comprises the steps of:

(a) contacting an agent with an isolated protein encoded by an ORF of the present invention, or nucleic acid of the invention; and

(b) determining whether the agent binds to said protein or said nucleic acid.

In general, therefore, such methods for identifying compounds that bind to a polynucleotide of the invention can comprise contacting a compound with a polynucleotide of the invention for a time sufficient to form a polynucleotide/compound complex, and detecting the complex, so that if a polynucleotide/compound complex is detected, a compound that binds to a polynucleotide of the invention is identified.

Likewise, in general, therefore, such methods for identifying compounds that bind to a polypeptide of the invention can comprise contacting a compound with a polypeptide of the invention for a time sufficient to form a polypeptide/compound complex, and detecting the complex, so that if a polypeptide/compound complex is detected, a compound that binds to a polynucleotide of the invention is identified.

Methods for identifying compounds that bind to a polypeptide of the invention can also comprise contacting a compound with a polypeptide of the invention in a cell for a time sufficient to form a polypeptide/compound complex, wherein the complex drives expression of a receptor gene sequence in the cell, and detecting the complex by detecting reporter gene sequence expression, so that if a polypeptide/compound complex is detected, a compound that binds a polypeptide of the invention is identified.

Compounds identified via such methods can include compounds which modulate the activity of a polypeptide of the invention (that is, increase or decrease its activity, relative to activity observed in the absence of the compound). Alternatively, compounds identified via such methods can include compounds which modulate the expression of a polynucleotide of the invention (that is, increase or decrease expression relative to expression levels observed in the absence of the compound). Compounds, such as compounds identified via the methods of the invention, can be tested using standard assays well known to those of skill in the art for their ability to modulate activity/expression.

The agents screened in the above assay can be, but are not limited to, peptides, carbohydrates, vitamin derivatives, or other pharmaceutical agents. The agents can be selected and screened at random or rationally selected or designed using protein modeling techniques.

For random screening, agents such as peptides, carbohydrates, pharmaceutical agents and the like are selected at random and are assayed for their ability to bind to the protein encoded by the ORF of the present invention. Alternatively, agents may be rationally selected or designed. As used herein, an agent is said to be “rationally selected or designed” when the agent is chosen based on the configuration of the particular protein. For example, one skilled in the art can readily adapt currently available procedures to generate peptides, pharmaceutical agents and the like, capable of binding to a specific peptide sequence, in order to generate rationally designed antipeptide peptides, for example see Hurby et al., Application of Synthetic Peptides: Antisense Peptides,” In Synthetic Peptides, A User's Guide, W. H. Freeman, NY (1992), pp. 289-307, and Kaspczak et al., Biochemistry 28:9230-8 (1989), or pharmaceutical agents, or the like.

In addition to the foregoing, one class of agents of the present invention, as broadly described, can be used to control gene expression through binding to one of the ORFs or EMFs of the present invention. As described above, such agents can be randomly screened or rationally designed/selected. Targeting the ORF or EMF allows a skilled artisan to design sequence specific or element specific agents, modulating the expression of either a single ORF or multiple ORFs which rely on the same EMF for expression control. One class of DNA binding agents are agents which contain base residues which hybridize or form a triple helix formation by binding to DNA or RNA. Such agents can be based on the classic phosphodiester, ribonucleic acid backbone, or can be a variety of sulfliydryl or polymeric derivatives which have base attachment capacity.

Agents suitable for use in these methods preferably contain 20 to 40 bases and are designed to be complementary to a region of the gene involved in transcription (triple helix—see Lee et al., Nucl. Acids Res. 6:3073 (1979); Cooney et al., Science 241:456 (1988); and Dervan et al., Science 251:1360 (1991)) or to the mRNA itself (antisense—Okano, J. Neurochem. 56:560 (1991); Oligodeoxynucleotides as Antisense Inhibitors of Gene Expression, CRC Press, Boca Raton, Fla. (1988)). Triple helix-formation optimally results in a shut-off of RNA transcription from DNA, while antisense RNA hybridization blocks translation of an mRNA molecule into polypeptide. Both techniques have been demonstrated to be effective in model systems. Information contained in the sequences of the present invention is necessary for the design of an antisense or triple helix oligonucleotide and other DNA binding agents.

Agents which bind to a protein encoded by one of the ORFs of the present invention can be used as a diagnostic agent. Agents which bind to a protein encoded by one of the ORFs of the present invention can be formulated using known techniques to generate a pharmaceutical composition.

3.19 Use of Nucleic Acids as Probes

Another aspect of the subject invention is to provide for polypeptide-specific nucleic acid hybridization probes capable of hybridizing with naturally occurring nucleotide sequences. The hybridization probes of the subject invention may be derived from any of the nucleotide sequences SEQ ID NOs: 1-948. Because the corresponding gene is only expressed in a limited number of tissues, a hybridization probe derived from of any of the nucleotide sequences SEQ ID NOs: 1-948 can be used as an indicator of the presence of RNA of cell type of such a tissue in a sample.

Any suitable hybridization technique can be employed, such as, for example, in situ hybridization. PCR as described in U.S. Pat. Nos. 4,683,195 and 4,965,188 provides additional uses for oligonucleotides based upon the nucleotide sequences. Such probes used in PCR may be of recombinant origin, may be chemically synthesized, or a mixture of both. The probe will comprise a discrete nucleotide sequence for the detection of identical sequences or a degenerate pool of possible sequences for identification of closely related genomic sequences.

Other means for producing specific hybridization probes for nucleic acids include the cloning of nucleic acid sequences into vectors for the production of mRNA probes. Such vectors are known in the art and are commercially available and may be used to synthesize RNA probes in vitro by means of the addition of the appropriate RNA polymerase as T7 or SP6 RNA polymerase and the appropriate radioactively labeled nucleotides. The nucleotide sequences may be used to construct hybridization probes for mapping their respective genomic sequences. The nucleotide sequence provided herein may be mapped to a chromosome or specific regions of a chromosome using well known genetic and/or chromosomal mapping techniques. These techniques include in situ hybridization, linkage analysis against known chromosomal markers, hybridization screening with libraries or flow-sorted chromosomal preparations specific to known chromosomes, and the like. The technique of fluorescent in situ hybridization of chromosome spreads has been described, among other places, in Verma et al (1988) Human Chromosomes: A Manual of Basic Techniques, Pergamon Press, New York N.Y.

Fluorescent in situ hybridization of chromosomal preparations and other physical chromosome mapping techniques may be correlated with additional genetic map data. Examples of genetic map data can be found in the 1994 Genome Issue of Science (265:1981f). Correlation between the location of a nucleic acid on a physical chromosomal map and a specific disease (or predisposition to a specific disease) may help delimit the region of DNA associated with that genetic disease. The nucleotide sequences of the subject invention may be used to detect differences in gene sequences between normal, carrier or affected individuals.

3.20 Preparation of Support Bound Oligonucleotides

Oligonucleotides, i.e., small nucleic acid segments, may be readily prepared by, for example, directly synthesizing the oligonucleotide by chemical means, as is commonly practiced using an automated oligonucleotide synthesizer.

Support bound oligonucleotides may be prepared by any of the methods known to those of skill in the art using any suitable support such as glass, polystyrene or Teflon. One strategy is to precisely spot oligonucleotides synthesized by standard synthesizers. Inmobilization can be achieved using passive adsorption (Inouye & Hondo, (1990) J. Clin. Microbiol. 28(6) 1469-72); using UV light (Nagata et al., 1985; Dahlen et al., 1987; Morrissey & Collins, (1989) Mol. Cell Probes 3(2) 189-207) or by covalent binding of base modified DNA (Keller et al., 1988; 1989); all references being specifically incorporated herein.

Another strategy that may be employed is the use of the strong biotin-streptavidin interaction as a linker. For example, Broude et a. (1994) Proc. Natl. Acad. Sci. USA 91(8) 3072-6, describe the use of biotinylated probes, although these are duplex probes, that are immobilized on streptavidin-coated magnetic beads. Streptavidin-coated beads may be purchased from Dynal, Oslo. Of course, this same linking chemistry is applicable to coating any surface with streptavidin. Biotinylated probes may be purchased from various sources, such as, e.g., Operon Technologies (Alameda, Calif.).

Nunc Laboratories (Naperville, Ill.) is also selling suitable material that could be used. Nunc Laboratories have developed a method by which DNA can be covalently bound to the microwell surface termed Covalink NH. CovaLink NH is a polystyrene surface grafted with secondary amino groups (>NH) that serve as bridge-heads for further covalent coupling. CovaLink Modules may be purchased from Nunc Laboratories. DNA molecules may be bound to CovaLink exclusively at the 5′-end by a phosphoramidate bond, allowing immobilization of more than 1 pmol of DNA (Rasmussen et al., (1991) Anal. Biochem. 198(1) 138-42).

The use of CovaLink NH strips for covalent binding of DNA molecules at the 5′-end has been described (Rasmussen et al., (1991). In this technology, a phosphoramidate bond is employed (Chu et al., (1983) Nucleic Acids Res. 11(8) 6513-29). This is beneficial as immobilization using only a single covalent bond is preferred. The phosphoramnidate bond joins the DNA to the CovaLink NH secondary amino groups that are positioned at the end of spacer arms covalently grafted onto the polystyrene surface through a 2 nm long spacer arm. To link an oligonucleotide to CovaLink NH via an phosphoramidate bond, the oligonucleotide terminus must have a 5′-end phosphate group. It is, perhaps, even possible for biotin to be covalently bound to CovaLink and then streptavidin used to bind the probes.

More specifically, the linkage method includes dissolving DNA in water (7.5 ng/ul) and denaturing for 10 min. at 95° C. and cooling on ice for 10 min. Ice-cold 0.1 M 1-methylimidazole, pH 7.0 (1-MeIm 7 ), is then added to a final concentration of 10 mM 1-MeIm 7 . A ss DNA solution is then dispensed into CovaLink NH strips (75 ul/well) standing on ice.

Carbodiimide 0.2 M 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide (EDC), dissolved in 10 mM 1-MeIm 7 , is made fresh and 25 ul added per well. The strips are incubated for 5 hours at 50° C. After incubation the strips are washed using, e.g., Nunc-Immuno Wash; first the wells are washed 3 times, then they are soaked with washing solution for 5 min., and finally they are washed 3 times (where in the washing solution is 0.4 N NaOH, 0.25% SDS heated to 50° C.).

It is contemplated that a further suitable method for use with the present invention is that described in PCT Patent Application WO 90/03382 (Southern & Maskos), incorporated herein by reference. This method of preparing an oligonucleotide bound to a support involves attaching a nucleoside 3′-reagent through the phosphate group by a covalent phosphodiester link to aliphatic hydroxyl groups carried by the support. The oligonucleotide is then synthesized on the supported nucleoside and protecting groups removed from the synthetic oligonucleotide chain under standard conditions that do not cleave the oligonucleotide from the support. Suitable reagents include nucleoside phosphoramidite and nucleoside hydrogen phosphorate.

An on-chip strategy for the preparation of DNA probe for the preparation of DNA probe arrays may be employed. For example, addressable laser-activated photodeprotection may be employed in the chemical synthesis of oligonucleotides directly on a glass surface, as described by Fodor et al. (1991) Science 251(4995) 767-73, incorporated herein by reference. Probes may also be immobilized on nylon supports as described by Van Ness et al. (1991) Nucleic Acids Res. 19(12) 3345-50; or linked to Teflon using the method of Duncan & Cavalier (1 988) Anal. Biochem. 169(1) 104-8; all references being specifically incorporated herein.

To link an oligonucleotide to a nylon support, as described by Van Ness et al. (1991), requires activation of the nylon surface via alkylation and selective activation of the 5′-amine of oligonucleotides with cyanuric chloride.

One particular way to prepare support bound oligonucleotides is to utilize the light-generated synthesis described by Pease et al., (1994) PNAS USA 91(11) 5022-6, incorporated herein by reference). These authors used current photolithographic techniques to generate arrays of immobilized oligonucleotide probes (DNA chips). These methods, in which light is used to direct the synthesis of oligonucleotide probes in high-density, miniaturized arrays, utilize photolabile 5′-protected N-acyl-deoxynucleoside phosphoramidites, surface linker chemistry and versatile combinatorial synthesis strategies. A matrix of 256 spatially defined oligonucleotide probes may be generated in this manner.

3.21 Preparation of Nucleic Acid Fragments

The nucleic acids may be obtained from any appropriate source, such as cDNAs, genomic DNA, chromosomal DNA, iicrodissected chromosome bands, cosmid or YAC inserts, and RNA, including mRNA without any amplification steps. For example, Sambrook et al. (1989) describes three protocols for the isolation of high molecular weight DNA from mammalian cells (p. 9.14-9.23).

DNA fragments may be prepared as clones in M13, plasmid or lambda vectors and/or prepared directly from genomic DNA or cDNA by PCR or other amplification methods. Samples may be prepared or dispensed in multiwell plates. About 100-1000 ng of DNA samples may be prepared in 2-500 ml of final volume.

The nucleic acids would then be fragmented by any of the methods known to those of slill in the art including, for example, using restriction enzymes as described at 9.24-9.28 of Sambrook et al. (1989), shearing by ultrasound and NaOH treatment.

Low pressure shearing is also appropriate, as described by Schriefer et al. (1990) Nucleic Acids Res. 18(24) 7455-6, incorporated herein by reference). In this method, DNA samples are passed through a small French pressure cell at a variety of low to intermediate pressures. A lever device allows controlled application of low to intermediate pressures to the cell. The results of these studies indicate that low-pressure shearing is a useful alternative to sonic and enzymatic DNA iragmentation methods.

One particularly suitable way for fragmenting DNA is contemplated to be that using the two base recognition endonuclease, CviJI, described by Fitzgerald et al. (1992) Nucleic Acids Res. 20(14) 3753-62. These authors described an approach for the rapid fragmentation and fractionation of DNA into particular sizes that they contemplated to be suitable for shotgun cloning and sequencing.

The restriction endonuclease CviJI normally cleaves the recognition sequence PuGCPy between the G and C to leave blunt ends. Atypical reaction conditions, which alter the specificity of this enzyme (CviJI**), yield a quasi-random distribution of DNA fragments form the small molecule pUC19 (2688 base pairs). Fitzgerald et al. (1992) quantitatively evaluated the randomness of this fragmentation strategy, using a CviJI** digest of pUC19 that was size fractionated by a rapid gel filtration method and directly ligated, without end repair, to a lac Z minus M13 cloning vector. Sequence analysis of 76 clones showed that CviJI** restricts pyGCPy and PuGCPu, in addition to PuGCPy sites, and that new sequence data is accumulated at a rate consistent with random fragmentation.

As reported in the literature, advantages of this approach compared to sonication and agarose gel fractionation include: smaller amounts of DNA are required (0.2-0.5 ug instead of 2-5 ug); and fewer steps are involved (no preligation, end repair, chemical extraction, or agarose gel electrophoresis and elution are needed

Irrespective of the manner in which the nucleic acid fragments are obtained or prepared, it is important to denature the DNA to give single stranded pieces available for hybridization. This is achieved by incubating the DNA solution for 2-5 minutes at 80-90° C. The solution is then cooled quickly to 2° C. to prevent renaturation of the DNA fragments before they are contacted with the chip. Phosphate groups must also be removed from genomic DNA by methods known in the art.

3.22 Preparation of DNA Arrays

Arrays may be prepared by spotting DNA samples on a support such as a nylon membrane. Spotting may be performed by using arrays of metal pins (the positions of which correspond to an array of wells in a microtiter plate) to repeated by transfer of about 20 nl of a DNA solution to a nylon membrane. By offset printing, a density of dots higher than the density of the wells is achieved. One to 25 dots may be accommodated in 1 mm 2 , depending on the type of label used. By avoiding spotting in some preselected number of rows and columns, separate subsets (subarrays) may be formed. Samples in one subarray may be the same genomic segment of DNA (or the same gene) from different individuals, or may be different, overlapped genomic clones. Each of the subarrays may represent replica spotting of the same samples. In one example, a selected gene segment may be amplified from 64 patients. For each patient, the amplified gene segment may be in one 96-well plate (all 96 wells containing the same sample). A plate for each of the 64 patients is prepared. By using a 96-pin device, all samples may be spotted on one 8×12 cm membrane. Subarrays may contain 64 samples, one from each patient. Where the 96 subarrays are identical, the dot span may be 1 mm 2 and there may be a 1 mm space between subarrays.

Another approach is to use membranes or plates (available from NUNC, Naperville, Ill.) which may be partitioned by physical spacers e.g. a plastic grid molded over the membrane, the grid being similar to the sort of membrane applied to the bottom of multiwell plates, or hydrophobic strips. A fixed physical spacer is not preferred for imaging by exposure to flat phosphor-storage screens or x-ray films.

The present invention is illustrated in the following examples. Upon consideration of the present disclosure, one of skill in the art will appreciate that many other embodiments and variations may be made in the scope of the present invention. Accordingly, it is intended that the broader aspects of the present invention not be limited to the disclosure of the following examples. The present invention is not to be limited in scope by the exemplified embodiments which are intended as illustrations of single aspects of the invention, and compositions and methods which are functionally equivalent are within the scope of the invention. Indeed, numerous modifications and variations in the practice of the invention are expected to occur to those skilled in the art upon consideration of the present preferred embodiments. Consequently, the only limitations which should be placed upon the scope of the invention are those which appear in the appended claims.

All references cited within the body of the instant specification are hereby incorporated by reference in their entirety.

4.0 EXAMPLES

4.1 Example 1

Novel Nucleic Acid Sequences Obtained from Various Libraries

A plurality of novel nucleic acids were obtained from cDNA libraries prepared from various human tissues and in some cases isolated from a genomic library derived from human chromosome using standard PCR, SBH sequence signature analysis and Sanger sequencing techniques. The inserts of the library were amplified with PCR using primers specific for the vector sequences which flank the inserts. Clones from cDNA libraries were spotted on nylon membrane filters and screened with oligonucleotide probes (e.g., 7-mers) to obtain signature sequences. The clones were clustered into groups of similar or identical sequences. Representative clones were selected for sequencing.

In some cases, the 5′ sequence of the amplified inserts was then deduced using a typical Sanger sequencing protocol. PCR products were purified and subjected to fluorescent dye terminator cycle sequencing. Single pass gel sequencing was done using a 377 Applied Biosystems (ABI) sequencer to obtain the novel nucleic acid sequences. In some cases RACE (Random Amplification of cDNA Ends) was performed to further extend the sequence in the 5′ direction.

4.2 Example 2

Novel Nucleic Acids

The novel nucleic acids of the present invention of the invention were assembled from sequences that were obtained from a cDNA library by methods described in Example 1 above, and in some cases sequences obtained from one or more public databases, The nucleic acids were assembled using an EST sequence as a seed. Then a recursive algorithm was used to extend the seed EST into an extended assemblage, by pulling additional sequences from different databases (i.e., Hyseq's database containing EST sequences, dbEST version 119, gb pri 119, and UniGene version 119) that belong to this assemblage. The algorithm terminated when there was no additional sequences from the above databases that would extend the assemblage. Inclusion of component sequences into the assemblage was based on a BLASTN hit to the extending assemblage with BLAST score greater than 300 and percent identity greater than 95%.

Using PHRAP (Univ. of Washington) or CAP4 (Paracel), a full length gene cDNA sequence and its corresponding protein sequence were generated from the assemblage. Any frame shifts and incorrect stop codons were conrected by hand editing. During editing, the sequence was checked using FASTY and/or BLAST against Genbank (i.e., dbEST version 121, gb pri 121, UniGene version 121, Genpept release 121). Other computer programs which may have been used in the editing process were phredPhrap and Consed (University of Washington) and ed-ready, ed-ext and cg-zip-2 (Hyseq, Inc.). The full-length nucleotide and amino acid sequences, including splice variants resulting from these procedures are shown in the Sequence Listing as SEQ ID NO: 1-948.

Table 1 shows the various tissue sources of SEQ ID NO: 1-948.

The homology for SEQ ID NO: 1-948 were obtained by a BLASTP version 2.0al 19MP-WashU search against Genpept release 120 and the amino acid version of Geneseq released on Oct. 26, 2000, using BLAST algorithm. The results showed homologues for SEQ ID NO: 1-948 from Genpept. The homologues with identifiable functions for SEQ ID NO: 1-948 are shown in Table 2 below.

Using eMatrix software package (Stanford University, Stanford, Calif.) (Wu et al., J. Comp. Biol., Vol. 6 pp. 219-235 (1999) herein incorporated by reference), all the sequences were examined to determine whether they had identifiable signature regions. Table 3 shows the signature region found in the indicated polypeptide sequences, the description of the signature, the eMatrix p-value(s) and the position(s) of the signature within the polypeptide sequence.

Using the pFam software program (Sonnhammer et al., Nucleic Acids Res., Vol. 26(1) pp. 320-322 (1998) herein incorporated by reference) all the polypeptide sequences were examined for domains with homology to certain peptide domains. Table 4 shows the name of the domain found, the description, the p-value and the pFam score for the identified domain within the sequence.

The nucleotide sequence within the sequences that codes for signal peptide sequences and their cleavage sites can be determined from using Neural Network SignalP V1.1 program (from Center for Biological Sequence Analysis, The Technical University of Denmark). The process for identifying prokaryotic and eukaryotic signal peptides and their cleavage sites are also disclosed by Henrik Nielson, Jacob Engelbrecht, Soren Brunak, and Gunnar von Heijne in the publication “Identification of prokaryotic and eukaryotic signal peptides and prediction of their cleavage sites” Protein Engineering, Vol. 10, no. 1, pp. 1-6 (1997), incorporated herein by reference. A maximum S score and a mean S score, as described in the Nielson et as reference, was obtained for the polypeptide sequences. Table 5 shows the position of the signal peptide in each of the polypeptides and the maximum score and mean score associated with that signal peptide.

TABLE 1
		HYSEQ
TISSUE	LIBRARY/	LIBRARY
ORIGIN	RNA SOURCE	NAME	SEQ ID NOS:
adult brain	GIBCO	AB3001	16-17 19 40 66 92-94 97 124 131 134 163
			186 188 208 213 231 268-270 284 288 295
			297 299 311 315-325 340 373 387 396 407
			429 469 489 495 498-499 533 542 545 562
			568 587 589 618-619 643 664 687-688 694-
			695 730 748 836 876 882 884 902 925-926
			948
adult brain	GIBCO	ABD003	2 22-24 29 33 43 45 50-51 66 71 75 77 82
			87-88 91-92 95 131 140 157 179 188-192 200
			208 213 220 225 247 252 257 261 263-265
			277 284 288 295 299 301 315-325 355-356
			373 387-389 392 395-396 407 423 431 443-
			444 450-451 457 459 468 476 489 495 499-
			500 514 520-522 532-533 541-542 545-546
			557-558 562 564 576-577 581-583 588-589
			591 595 597 599 601 610 619 631-632 639
			643-644 654-655 658-660 664 667 676 682
			687-688 693 696 700 704 711 713-714 746
			758 765-766 774-775 780 800 802 804 807
			810 827 829 834 842 850 854-855 866 870-
			871 878 892-893 897 899 910 916 920-921
			929 931-932 934
adult brain	Clontech	ABR001	12 51 87 142 169 178 180 245 263 286 288
			290 295 304 308 311-313 375 379-380 403
			425 428 431 458 486 499 503 512 557-558
			567-568 606 610 641 651 695 704 730 741
			754 766 810 822 827 841 850 864 871 884
			897 917 920 925-927 934 946
adult brain	Clontech	ABR006	2 14-15 22-23 29 32-33 49 66-68 83 99 111-
			112 115 129 131 142 147 153 157 163 169
			189-192 200 205 207 212-214 218 221 229
			234 256-257 263 272 276 279 282 292-299
			301 311 315 340-343 349 376-377 383-386
			388 403 405 407 410 425 438 453-454 460
			463 469 474 489 495 499-500 511 522 531-
			532 539 541 545-546 551 556 563 565 571
			579-583 591 594 606 626 628 631-632 643
			647 651 678 684 691-692 700 717 721 726
			730 732 741 744 754 757 769 772 774 782
			788 793 810 820 827-828 853 867 869 875
			879 897 913 921-922 925-926 933-934 939-
			941 947
adult brain	Clontech	ABR008	1-2 9-10 13 16-18 23 27-28 30-32 37 39 42-
			43 46 49-51 66 70 76 80 83 86-87 95-97 109
			111-112 116-117 124 130-131 133-134 136-
			137 141-142 146-147 152-157 160 162 169
			171 179 184 189-192 195 200-201 206 211-
			212 216-218 239 247-248 250 252 254-258
			261-263 271-272 276 278 282 288 293-295
			297 300 302 307 309 311 314-326 328 333
			337-341 343 347 349 351-354 358 360-361
			367 374 376-378 381 384 388-390 393 395-
			396 400-403 405 407 409 411 414 418-420
			422 427-429 433 438 440-441 445-447 450
			453-455 458 460-461 463 466-470 474 476
			486 491-493 496 498-500 507 511 514 520-
			521 525 527-529 531-532 534-535 542 546
			548-549 551-552 557-558 560 562 564-566
			568 571-572 578-583 586-587 590-591 594
			599 602 606 618-619 621 626 629 631-634
			643-644 647 651 656-660 664 670 672 677
			680 684 687-688 691-695 697 706 709-710
			712-714 716-718 721-722 724-725 727-728
			730 733 740-741 745 751-752 754 761 765
			774 777-779 787 790 792-793 799 801-804
			808 810 812 820 822 824 827 831-832 834
			836 845 850 858-861 868-869 871-672 875-
			876 883 887 891 897 900 904 907 910 913
			917-920 925-927 929 931-934 938-941 946-
			947
adult brain	Clontech	ABR011	51 133 810 892
adult brain	BioChain	ABR012	140 208 311 748 810
adult brain	Invitrogen	ABR013	51 245 311 316-325 436 717 810 936
adult brain	Invitrogen	ABR014	2 51 65 84 86 134 311 316-325 384 422 445
			460 503 525 564 634 651 721 794 804 810
			922
adult brain	Invitrogen	ABR015	37 134 263 272 277 294 311 443 467 500 514
			582-583 619 651 694 850 871-872 883 888
			936
adult brain	Invitrogen	ABR016	19 22 57 134 188 233 271 277 299 373 440
			444 459 469 514 640 717 882 890 920
adult brain	Invitrogen	ABT004	1-2 18 28 51 55-57 67 87-88 115 119 137-
			139 142 163 200 204 213 218 257 263 271
			282 288 299 301 311 341 358 370 378 402
			407 422-423 427 458 460 463 499 504 534-
			535 551 557-558 571 586 605-606 610 618
			627-628 640 643 680 687 691-692 697 701-
			702 715 719-721 725 727 753-754 758 771
			782 810 827 859 871-872 881 913 920 925-
			926 938-941 944 946
cultured	Stratagene	ADP001	2 43 51 73 76 88 97 142 166 181 186 188
preadipocytes			208 257 262-263 267-270 282 311 316-325
			383 386 427-429 459 463 465 493 507 514
			522 545 552 572 643 651 667 700 721 740
			754 758 778 795 872 881 883 888 947
adrenal gland	Clontech	ADR002	3-6 10-11 13 16 20-21 24 27-28 33 38 48-49
			51 53-54 58 66-67 75 88 97 99 124-125 130
			140 157-158 179 188 197-198 200 212-214
			216 218 224 229 231 237 257 267 279 281-
			282 288 302 311 326 362 376-377 381 383
			396 398-403 429 443 453-454 456 459-460
			474 489 515 526 531-532 540 545 550 559
			564 568 577 581 586 589 599 605 610 613
			631-632 643 648 651 667 670 672 681 684
			699 703 706 708 717-718 734 736 751 779
			785-786 795 813 817 837 871 876 887-888
			897 904 907 916 921 924-926 948
adult heart	GIBCO	AHR001	1-2 5-6 14-18 20-21 23 28 32 37 41 45 51
			53 55-56 62 66 69-70 80-81 85 87 91 97 107
			120-121 124 134 140-141 156 163 165-166
			172 188-192 195 197-198 200 208 213 216
			221 229 231 235 261-265 267 271 276 284
			288 302 305 308 311 316-325 328 333-334
			337-338 347 368-369 373 376-377 379-380
			389 396 420 440 445 453-454 459-460 465
			468 478 483-484 489 491-493 495 501 504
			507 514 524 529 533 539 541-543 545 549
			552-553 564 566 568 574 577 581-583 587
			589-591 596 599 602 605 608-609 618-619
			623 625 629-632 643 645 647 651 664 672
			676 678 683-684 707 714 716-717 732 735
			740 743-744 751 754 757 765 775 778 784-
			786 788 807-808 810 826 828-829 842 850
			860 876 878-880 890 894 897 899 902 916
			923-927 933 939-941
adult kidney	GIBCO	AKD001	1-2 5-6 13 16-17 19-23 26 28 33 38-39 43
			45 48-51 55-57 60 66-67 69-73 79 82-83 87
			90 94 96-97 100 103 126 131 134 140 148-
			149 157 163 166 179 184 186 188-192 200-
			203 213-216 220-221 224 226-229 232 235
			245 252 257 261-263 268-270 272-274 276-
			277 279 282 288 290 294 299 308 311 316-
			325 332 335 339-340 358 360-363 373 375
			379-380 386 388-389 392 395-396 402 413
			421 423-424 428-429 431 436 440 444 450
			454 457 459-460 468-469 476 489 492-493
			499 504 511 513-514 520-521 524-526 531
			533 538-542 544-547 552 564 567-568 574
			577-578 582-583 590-591 595-596 598 602
			607 610 613 618-619 622 631-632 639-642
			644 647 651 654-655 658-659 664 667-669
			673 678 680-682 684 687 689 693 696 706-
			707 712 714-715 717-718 721 729-731 734-
			736 740 744 748 754 760 771 774 782 784
			789 795 807 809-810 819 825 834 836-837
			842 850 859 870 872 876 878-879 884 887
			890 895 897-899 902 905 910 919-921 925-
			926 933 936 944
adult kidney	Invitrogen	AKT002	1 14-15 28 30 35-37 53-54 73 88 112 114
			129 134 137 140 149 157 166 172 186-188
			191-192 203-204 213 235 245 257 262-263
			266 268-270 273-274 288-289 297 299 302
			310-313 315-325 335 340 358 373 378-381
			395 413 423 441 450 453 456 459-460 470
			477 491-494 500 513 540 542 545 554 556
			564 567 587 591 619 622 627 633 643 668-
			669 677 684 689 693 701-702 704 714 729-
			730 754 758 760 777 781 785-786 788-789
			807 836-837 840 849-850 872 876 881 890
			895 905-906 913 923 925-926 931-933 944
adult lung	GIBCO	ALG001	5-6 16 28 38 51 74 97 122 124 134 140 163
			188-192 200 218 221 262-263 268-272 294
			311 316-325 379-380 429 463 468 493 511
			520-522 537-538 542 545 568-569 595 622
			643-644 664 667 711 714 721 730 754 775
			850 860 863 879 887 897 925-926 944
lymph node	Clontech	ALN001	43 98 131 140 163 188 221 245 277 299 311
			491 515 546 564 593 603 610 615 630 682
			694 707 717 800 831 850 878 880 936 939-
			941 947
young liver	GIBCO	ALV001	3-4 17 20-21 32 43 55-56 70 100 134 137
			163 172 174 179 186 188-192 200 213 216
			219 221 229 232 252 275 301 311 315-325
			378 381 392 441 459-460 497 499-500 514
			524 526 533 539 550 568 571 588-589 595
			619 622 631-632 642 658-659 664 677 680
			693 700 707 713 719 743 754 757-758 766
			807 834 863 867 876 884 887 904 907
adult liver	Invitrogen	ALV002	5-6 28 35-36 52 54 70 72 86-87 103 112 127
			134 140 159 179 188 200-201 213 218-219
			225 239-240 257 263 271 275 311 315 367
			373 388 392 444 459-460 464 468 497-499
			512 527 532 542 545 562 599 605 629 640
			657 680 684 687-688 706 713 715 717-718
			721 742 754 758 771 791-793 818 829 843
			854-855 871 878-879 887 921 933-934
adult liver	Clontech	ALV003	159 179 189-192 201 219 257 349 392 568
			664 753 796 887 934
adult ovary	Invitrogen	AOV001	2-7 10 13 18-22 25 27-28 30-31 33 38-39
			41-43 45 48 50-51 53-56 62 66-67 69 72 74-
			75 80 83 85-87 93 95 99-101 107 112-115
			120 124-126 129 131 134-137 140 142-143
			147-148 162-163 172 178 188 191-192 200
			204 208 212-213 220-221 225 229 231 235
			237 246-247 252-253 258 261-262 264-265
			267-272 276-279 282 287-288 290 293-294
			299 307-308 311 316-327 332 337-338 340-
			342 349 360-362 373-374 379-381 386 388-
			389 393 396 399 403 413-414 423 425 427
			429 431 441 444-445 450 452 454-455 457
			459-460 462 467-470 475 477 483-484 489
			491-493 495-496 500 504-505 507 515 518-
			519 522-523 527-529 531 533 537-540 542
			545-546 548 551-552 555 564 568 570-571
			577 579 581-583 589-596 599-600 605 610
			613 615-616 619 623 625-627 630-636 639
			641-644 647 649 651 654-655 664 668-669
			672 676 678 680-682 684 687 694 701-704
			706-707 715-717 721-722 727 729 731-732
			734-735 738 740 743 748-749 753-754 758
			764 771 775 777-782 784 800 802 807 821-
			822 824 828 834 836-837 842 846-847 850
			860-863 866 870-871 876-880 882 884 887
			890-891 897 899 901 906 910 913 920-921
			923-924 933 939-941 944 947
adult placenta	Clontech	APL001	34 68 102 263 444 493 520-521 534-535 689
			706 754 797-798
placenta	Invitrogen	APL002	2 14-15 43 55-56 66-67 134 184 213 221 229
			252 257 263 277 287 394 443 529 532 618
			622 684 742 754 810 829 883 902
adult spleen	GIBCO	ASP001	2 14-15 20-22 29 38 43 48 51 53-56 65 67
			72 74 84 87 131-132 134 137 140 172 188-
			192 200 212 221 256 263 271 282 308 311
			316-325 343 383 389 423 436 441 443 459-
			460 467 469 495 499-500 505 514 520-522
			524 529 537 539 545 552 585 619 631-632
			639 643 664 673 707 723 735 742-744 758
			771 799 810 817 836 850 878 925-926 934
			936
testis	GIBCO	ATS001	1 3-4 14-16 28 31 45-46 66 85 90 95 97 103
			112 128-130 134 140 163 166 188 191-192
			199-200 213 226-228 261-265 267-271 284
			302 311 316-325 327 379-380 391 413 421
			428 444 454 457 459-460 467 491 493 495
			500 505 519 525 529 532 534-535 545 552
			556 566 568 575 596 599 613 616-617 647
			649 651 680 684 703 707 716 719 721 727
			734 738 740 744 748 758 765-766 774 777
			782 802 810 817 827-828 834 842 846-847
			850 862-863 871-872 878 880 892 901 916-
			917 921
adult bladder	Invitrogen	BLD001	5-6 8 20-21 28 72 91 122 126 130 166 188
			197 200 213-214 225 257 262 315-325 341
			409 486 491 572 593 622 650 673 691-692
			810 813 861 870 877 883 887 904
bone marrow	Clontech	BMD001	8 13-16 28 38 43 45-48 50-51 57 62-63 65
			67 84-85 97 100 104 118 122-124 131 134
			140 163 188 214 216 221 224 231 245 252
			261-263 268-270 273-274 279 288 290 311
			373 378 389-391 395 414 428 431 436 440-
			441 443 451 455 459-460 465 469-470 475
			495 497-498 502 507 514-519 529 537-538
			542 546 550 552 556 560-561 563-564 568
			576-577 580 587 589 596 601-602 610-613
			619-620 626 642-643 647 651 664 666 668
			676 678 681-682 684 696 704 706-707 715
			727 730 732-735 740 748 753 758 761 764
			771 775 780 794 800-801 830 834 836 842
			850 863 871-872 878-879 882 884 888 897
			900-901 904 910 921 923 929 934 947
bone marrow	Clontech	BMD002	1-2 5-6 10 13 16-21 27 31 38 42-43 46 57
			65-66 76 80 84 87 97 99 110 112 118 131
			134 137 140 145 161 163 165 172 195 206
			208 221 229 231 237 244 247 252 256 267-
			270 272 276 278-279 282 284 288 294 301
			304 307 311 316-327 333-334 337-338 345-
			347 352 360-361 368 373 376-378 381 383
			388 414 436 441 443 450 452 454-455 457
			469-470 483-484 486 490 498 516 519-521
			524 530-531 539 542-543 545-546 551 553
			555 559 564 571 576-577 580 585 591 594
			602 604-605 607-608 610-612 619-621 625-
			626 629 631-632 639-640 644 650-651 664-
			665 684 687-688 693 699 703 714 723-724
			727 733 735 740 742 745 748 750-752 754-
			755 777-780 784 787 794-795 802 809 817
			824 827 831-832 834 846-847 850-851 854-
			855 861 867 875 878 883 886 891 894 897
			900 902 910 914 919 921 925-926 929 936
			939-941 944
bone marrow	Clontech	BMD004	65
bone marrow	Clontech	BMD007	65 76 84 245 516
adult colon	Invitrogen	CLN001	19-21 53 55-57 72 88 133-134 168 213 245
			252 311-313 316-325 340 443 459 469 483-
			484 486 497 515-516 597 606 622 643 667
			676 706 718 742-743 753 766 829 833 872
			887 902 923 929
Mixture of 16	Various	CTL016	52 137 189-192 316-325 529 591
tissues-	Vendors*
mRNAs*
Mixture of 16	Various	CTL021	65 84 169 189-192 311 316-325 406 676 727
tissues-	Vendors*		782 850
mRNAs*
Mixture of 16	Various	CTL028	65
tissues-mRNAs*	Vendors*
adult cervix	BioChain	CVX001	3-4 14-16 20-23 25 33 42-43 45 48-50 54 57
			67 69 75 85 87 91 95-97 107 110 114 124
			126-127 131 134 137 140 150 157 163 165
			172 185-188 200 204 212-213 216 225 229
			245 252 257 261-263 266-270 276 282 288
			290 301-302 308 316-325 327 340 363-364
			372-373 378 383 388-392 394 396 409 413-
			414 421 428-429 438-440 443-444 454 456-
			457 459 463 467 475 486 489 493 495 507
			514-515 522 534-537 556 568 572 574 577
			582-583 587 594 600 608 610 613 622 626
			633 639 643 647-648 651 653 667 680 683
			685-686 693 696 703-704 706 711 721 723-
			725 727 730-731 734-735 742-743 748 754-
			757 762 771 776 785-786 788 794 800 802
			807 809-810 817 827 829 834-835 842 850
			857 860 862-863 868 870 873 876-877 879-
			880 884 887 891 897 904-905 910 916-917
			921 925-926 933 937 947
diaphragm	BioChain	DIA002	305 311
endothelial	Stratagene	EDT001	1-2 7-8 14-16 19-22 24 28-29 32-33 41 43
cells			45 51 57 61 74 83 87-88 97 105 112 116-117
			131 134 137 140 148 165 172 179 188-192
			197-198 208 212-213 220-221 225 229 231
			237 246 252 256-258 261-265 268-272 276-
			277 279 281-282 284 286 288 294 297 299
			302 307-308 311-313 326 334-335 340 355-
			356 358 360-361 364 375 383 386 389 392
			403 413 423-424 429 440 443 445 451 453
			455-456 459-460 462-463 465-466 468-470
			475 491 495 497-499 504 514 520-522 524-
			526 528 532-536 539-540 546 551-552 554
			556 564 566-567 571 574-577 581-583 587
			591-593 597-599 601 607 615 618 622 625
			633 639 641-644 651 667 677 680 684 691-
			692 701-702 704 716-717 720-721 726 732-
			733 735 743-744 754 758 765 785-786 795
			802 806 809 819 826 828-830 832 834 836
			846-847 850 867 871 877-878 890-891 897
			902 907 921 923 925-926 944 946
esophagus	BioChain	ESO002	188
fetal brain	Clontech	FBR001	33 49 51 126 134 197-198 264-265 360-361
			413 460 647 810 819 871
fetal brain	Clontech	FBR004	137 156 205 282 284 405 424 480 489 701-
			702 820 921
fetal brain	Clontech	FBR006	2 9-10 18-19 22 28 30-32 37 39-40 42-43
			46-47 49 57 66-67 76 80 83 96 109 112 116-
			117 120 124 131 133-134 136 142-143 146
			152 155 160 162 165 169 173 184 189-198
			200-201 205 215-216 238 244 248 254-255
			257-258 260-263 272-274 276-277 282 288
			293-294 307 309 311 314-328 343 347 351-
			352 354 357-358 360-361 373-375 378-381
			390 392 400-401 403 405 407 410-411 413
			420 424 429 445 450 452-453 458 460 463
			467-469 472 474 477 479 483-484 491 499
			507 520-521 525 527 529 531 533 538 545
			551 562 564 566 571 574 579 581-583 587
			591 599-600 604 606 611 626 629 631-632
			638 643 651 654-655 657-660 672-673 676-
			677 684 689 693-694 697 699 709 714-715
			717 720-721 732-733 735 744 748 751-752
			754 761 763 767 772 775 777-779 781 785-
			786 790 792 802 804 808 810 820 824 826
			838-840 850 858-860 864 866 872-873 881
			891-892 901-902 904 910-911 913 917-918
			920 925-926 933 939-941 946-947
fetal brain	Clontech	FBRs03	316-325 684
fetal brain	Invitrogen	FBT002	2-4 20-21 45 51 53 57 88 93 125-126 134
			166 184 186 188 200 213 224 263 276-278
			307 311 341 373 375 418-419 423 427 432
			450 452 459-460 470 492 498-499 507 514
			522 534-535 545 550-552 571 577 610 714
			721 743 754 795 827 861 866 872-873 887
			896 925-926 934 939-941 946 948
fetal heart	Invitrogen	FHR001	2-4 10 13 16-18 29 31-32 37-38 43 46 49 51
			53 55-56 67-68 75 80 85 87 97 115 120 137
			152 156 160-161 163 168-169 174 178 189-
			192 196 200 216 220 225 252 262 276-277
			282 288 301-302 305 311 315-325 333 343
			351 357-358 360-361 368-372 378 424 436
			440-441 445 453 460 469 478 483-484 495
			520-521 527 533 538 541-543 546 556 564-
			566 568 576 581 587 594-595 601-602 606
			609 612 615 633 638 640 643 653-655 664-
			665 672-673 677 684 691-693 697 704 707
			709 717 735 738 744 746 748-749 751-752
			754 761 777 779 781-782 785-786 797-798
			820 824 826 829 834 838 841-847 850 875
			877-878 893-894 897 901 910 913 925-927
			936 946
fetal kidney	Clontech	FKD001	8 14-15 32 43 50 68 96 106 126 131 134 140
			186 188 226-228 233 279 282 311 339 428
			440 450 456 468 552 618 651 700 726 735
			748 751 781 794 797-798 826 878 887 899
fetal kidney	Clontech	FKD002	50 83 96 131 134 143 163 172 193-194 201
			203 215 263 273-274 311 316-325 339 360-
			363 374 376-377 379-380 388 394 400-401
			403 407 425 440 451 454 493 525 536-538
			540-542 572 580 582-583 587 605-606 621
			631-632 647 673 689 706 709 714 726 735
			761 774 777 799 809 845-848 858 872 875
			878-879 882 895 918 927
fetal kidney	Invitrogen	FKD007	66 214
fetal lung	Clontech	FLG001	65 179 213 223 340 360-361 491 564 577 591
			627 646 650 712 715 744 758 939-941
fetal lung	Invitrogen	FLG003	49 54 75 97 137 148 152 188 197-199 213-
			214 225 240 256 288 316-325 369 378 392
			423 429 464 496 526 580-581 586 591 693
			706 726-727 766 878 913 925-926 939-941
fetal lung	Clontech	FLG004	388 921
fetal liver-	Columbia	FLS001	1-23 25-39 41-43 45-46 49-65 76 83-84 87-
spleen	University		88 91 94 97 100 112 118 122 126 129-130
			134 137 140 148 163 168 172 179 186 188-
			189 191-192 197-198 200-201 213-214 216
			221 225-229 231-232 235 242 252-253 256-
			258 262-271 277 284 287-288 297 299 307-
			309 311 315-326 330 355-356 360-362 370
			373 378-380 388 392 394 396 400-402 413
			426 428 436 440-441 443-447 450 454-455
			457 459-460 463-465 467-469 475 477-478
			489-509 511-514 519-521 525-527 529-535
			537 539 542 545 551-552 555-556 559 561
			567 569 571 576-577 580-583 586-587 589
			591-592 595 598-602 605 607 610 612-613
			618-619 623 625-626 631-632 638-640 642-
			644 646-647 649 651-652 654-655 667 673
			676 680-681 683-684 700 703-704 706 711
			715-718 720-721 726 732-735 740 742-744
			748 754 756-758 763-765 771 774-775 777-
			782 785-786 790 793-795 797-798 806-808
			815 818 824-826 829-830 834-835 837 841
			846-847 849-850 856 860-861 866 870-872
			876 878 881-883 888 894 897-898 902 905
			907 910 919 924-926 929 933 942 947
fetal liver-	Columbia	FLS002	1 3-4 11-12 14-17 20-23 26-29 32-34 38 41
spleen	University		43 45-47 49 51-52 55-62 65-67 76 83-85 87-
			88 90-91 95 97-99 104-105 112 114-117 126
			130 133 150 163 165 172 178 186-187 193-
			194 200-202 208 213 221 225 229 232-235
			244-246 248-253 256-257 262-265 267-271
			273-274 284 287 299 311 315 326 335 337-
			338 343 355-356 358 375-378 381 392 394
			400-402 414 416 426 428-429 440-442 444-
			447 453-455 457 459 461 464-465 467 476-
			477 483-484 489-490 492 495 497-500 504
			506-507 509 511-514 519 522 524 526 532-
			535 537 539-540 542 545 551-552 556 567
			569 574 576-577 581 589-590 592 599 601-
			602 605 607 610 612-613 619-620 625 627
			629 631-632 638 640-641 646 648-649 654-
			655 667 670 683-684 687-688 693 696 700
			703-706 713 716-721 726 734-735 740 742
			744 748 754 758 771 775 777-778 780-782
			785-787 790-792 794-799 801-802 806 808
			818 824 829 835 849-850 852-855 857 870-
			871 876 882-884 886 888 890 894 897-898
			900 902-903 907 919 921 923-926 929-930
			933 938-942
fetal liver-	Columbia	FLS003	30 34 67 85 88 89 130 172 188-189 191-192
spleen	University		213 229 231 257 311 315-325 329 331 335
			362 391 394 400-402 423 441 455 457 461
			476 498 500 511 523 531-532 537 542 576
			587 592 612-613 625 649 665-666 703 719
			731 733 740 744 771 775 777-778 787 797-
			798 819 824 826 850 854-856 861 863 870
			879 884 897 923 931-933 947
fetal liver	Invitrogen	FLV001	2 7 19 28 35-37 47 52 54-56 66 95 134 139
			179 188-192 200 213 218 263 272 288 294
			305 311 315 349 378-381 388 392 403 426
			443 454 459-460 469 496-499 514 527 529
			532 534-535 555 586 605 640 644 658-659
			673 680 687 698 713 715 720-721 723 726
			754 758 778 795 817-818 829 853 861 868
			870-871 897 903 933
fetal liver	Clontech	FLV002	52 189-192 219 297 308 335 364 378 427 828
fetal liver	Clontech	FLV004	2 19 28-29 37 39 49 52-53 55-56 62 65-66
			76 87 124 134 137 139 142 179 188 195 208
			216 219 244 252 263 268-270 272 277 287-
			288 294 303 305 311 315-325 339 355-356
			358 360-361 368 374 378 403 441 454-455
			460 477 483-484 497 514 520-521 542 553
			582-583 587 591 594 611-613 620-621 638-
			639 654-655 658-659 681 684 687-688 709
			721 730 738 744 752 754 781 793 802 813
			818 826 832 836 854-855 876 878 893 897
			900 910 924 933 944
fetal muscle	Invitrogen	FMS001	28 65 115 121 126 134 137 156 168 172-173
			181 213-214 225 263 267 305 340-341 360-
			361 440 459 516 534-535 543 564 586 606
			609-610 623 650 676 683 754 766 853 871
			886 894 930 934-935 948
fetal muscle	Invitrogen	FMS002	19-21 41 49 51 53 57 75 96 101 103 112 134
			136 156 171 184 188 191-192 212 216 250
			262 267 276 305 311 342 348-350 355-356
			360-361 374 392 403 411 415 423 425 457
			469 491 495 499 508 515 517 534-536 543
			546 564 566 576 580 582-583 587 594 599
			609 611 615 618 623 644 647 658-659 664
			668-669 677 683 691-692 696 703 735 743
			754 766 788 802 817 826 828 850 877-879
			894 910 925-926 935
fetal skin	Invitrogen	FSK001	3-7 18 24 27 29 35-37 51 53 55-56 66-67 76
			90 95 97 122 126 134 136-137 166-167 181
			188-192 208 213-214 224-225 245 250 252
			257 260 262 268-271 273-274 282 284 297
			302 312-313 315-326 341 367 373 375 378
			383 387-388 390 394 423 429 440-441 450
			454-455 457 459 463-464 470 472 475 486
			489 492 495 498-500 511 514 524 527 530-
			532 534-535 541-542 545 550 552 555-556
			565 571 586 589 591-592 602 604 606 610
			613 618 622-623 626 631-632 640 651 654-
			655 672-673 685-686 693 701-702 704 706
			717-718 720 723 727-728 744 754 762 764-
			766 768-771 795 809 814 821 824 827 843
			853 868 870-874 887-888 890 897 902 907
			925-926 928 930 933-934 939-941 944-945
			947
fetal skin	Invitrogen	FSK002	2 5-6 19 29 34 51 57 59 88 97 101 124 131
			134 143 163 166 172 189 191-192 196 212
			216 222-223 231 250 257 263 268-272 282
			284 287-288 294 297 299 302 304 310-311
			316-325 328 333 340 352 360-361 365-367
			372 379-380 388 390 400-401 403 410-411
			440 449-450 454 457 463 470 478 491 495
			500 505 515 520-521 524 532 534-535 541
			555 560 562 564 572 576 581 592 595 599
			611 622 626 630 636 640 642 650 664 677
			683 691-693 696 699 701-702 708-709 715
			721 723 728 735 744 747-748 750 754 766
			779 782 799 803 807 813 820 824 826 834
			846-847 867-868 872-874 878-879 890-891
			897 901 904 907 910 912 916 918 925-926
			933 944-945
fetal spleen	BioChain	FSP001	311 748
umbilical cord	BioChain	FUC001	1-2 29 32 46 67 83 87 94 134 136 140 148
			160 163 166 172 181 186-192 197-198 208
			213 216 225-231 237 252 261-265 267-270
			279 282 288 295 302 308 311 316-326 339-
			340 365 376-377 379-380 384 392-397 421
			423 428 433 440 445 452 459 461 463-464
			470 472 489 491 495 497 500 507 517-518
			522 525-526 528 534-535 540 545-546 556-
			558 564 566 568 571-572 577 592 599 601
			605 610 618 623 644 651 661 668-669 673
			678 680 685-686 696 706 709 718 735-736
			748 754 769 772-777 782 792 797-799 802
			807 809 815 817 824 850 854-855 870 876
			881 888 891 897 899 901 913 921 928 930-
			932
fetal brain	GIBCO	HFB001	2 12 16-17 19 23 27-28 32-33 39 41-45 49
			87-89 94 97 100 107 112 130-131 134 142
			157 163 172 188-192 200 216 224-225 231
			237 242 246 252 258 261 263-265 271 273-
			274 276-277 288 295 299 301 307 311 314-
			326 328 341 355-356 373 375 387 389 392
			395 424-425 431 438 445 450-452 457 459-
			460 468-469 475 489 491 495 500 504 511
			514 520-529 531 533 540-542 545 552 554
			557-558 566 576-577 579-584 587 591 596
			598-599 606 613 626 631-632 643 651 664
			668 673 676 680 693-694 696 703-704 716-
			717 721 727 735 738 740 744 748 757-758
			769 774 778 780-781 810 827-828 830 850
			869 871-872 876 878-879 884 890 892 897
			899 904 906-907 913 916 918 920 924 928
			934 938 946
macrophage	Invitrogen	HMP001	49 97 208 252 301 306 311 316-325 337-338
			345-346 416 512 522 572 670 716 743 785-
			786 802 888 919 923
infant brain	Columbia	IB2002	2-4 12 14-15 20-21 23-24 27 29 31-32 39 41
	University		46 48-49 51 53 55-56 66 75-76 86-88 93 95
			101 105 108-109 116-117 125 127 129 131
			136 145 163 166 170 180-181 186 188-189
			191-192 200-201 207-208 212-214 216 220
			224 229 231 245 247 252 257 259 264-265
			267 271 279 282 288 293 295 299-300 311
			314-326 337-338 340 349 367 373 375 388
			390-393 396 402 405 407 418-421 424 428-
			429 431 433 436 450 452-453 457 459 463
			468 489 495 496-500 507 511 522-524 526
			528-530 532 541-542 545-546 552 557-558
			562 564-566 571 577-583 587 589-591 599-
			601 606 608 613-614 619 631-632 647 654-
			655 658-659 667 676 684 691-693 696 700
			704 711 718 721 723 725 740-741 743-744
			748 754 775 777-778 780 788 792-793 795
			802 805 808 819 826-829 834 836 838 861
			863 869-870 875 879 881-882 884 887 890-
			891 893 897 902 920-921 925-926 934 938-
			941 946
infant brain	Columbia	IB2003	2 27 37 39 43 48-49 51 53 85-87 97 106 113
	University		124 126-127 131 142 166 170 188 200-201
			208 214-215 220 224 226-228 231 251 257
			263 267 271-272 279 288 293 299 311 314-
			326 337-339 349 360-361 367 386 392 397
			400-402 407 410 418-419 424-425 427 429
			452 454 460 475 489 495 497-500 507 522-
			523 525 529 532 539 542 545-546 551-552
			557-558 564-565 578 582-583 585 591 601
			606 625-626 631-633 643-644 673 690-693
			701-702 706 711 721 723 734 740-741 743-
			744 748 751 754 761 778 788 795 802 808
			819 826-827 829 837 843 869-871 875 878-
			880 884 896-897 902 920 933-934 946
infant brain	Columbia	IBM002	32 43 66 340 387 541 562 693 712 751 795
	University		829 871 920 925-927 929
infant brain	Columbia	IBS001	2 29 37 39 76 142 163 392 455 495 499 606
	University		681 741 754 778
lung,	Stratagene	LFB001	2-4 22 28 32-33 47 51 79 120 129 134 140
fibroblast			163 172 188 208 220-221 231 252 257 263
			276-277 284 307 375 378-380 396 423 428
			440 450 459 463 486 491 493 495 499 539
			571 591 601 607 613 615 618 625 639 651
			684 716-717 721 727 735 748 782 828 850
			870-871
lung tumor	Invitrogen	LGT002	2 5-8 13 16-17 29-31 35-39 43 46 57 67 72
			76 78 81 85 87 90 94 97 100 110 119 122
			130-131 134 137 140 146 149 167 172 174
			179 188 197-198 201 213 216 218 220-221
			223 231 245-246 251-252 256-257 262-263
			267-270 277 284 288 296 299 301-302 311
			316-325 340 354 373 379-380 388 392 395
			400-401 410 413 421 431 436 441-443 445
			451 455 457 460 463-464 467 469 475 478
			489 491 493 497 499 504 507 514 518-519
			524 529 534-535 537 542 545-546 548 552
			555 559 568 578 581-583 592 597 602-603
			605-607 613 615 619 621-622 636-637 642-
			643 646-647 654-655 679-681 684 687-689
			693 701-702 704 706 711 713 715-716 718
			727 732-734 738 748 753-754 757-758 760
			762 766 769 774 782 785-786 802 817 829
			834 850 853 859-860 866-867 870-871 878-
			879 887 890 899 902 904 910 917 923 925-
			926 936-937
lymphocytes	ATCC	LPC001	2 16 19-21 25 31-32 49 53 55-56 63 67 85-
			87 90 97 120 122 137 140 163 165 168 172
			188 197-198 215-216 221 229 231 236-238
			248 252 256 272-274 283-284 288 294 299
			316-326 343 368 374 378 395 423 431 454-
			455 467 469 476 478 491 495 498 505 512
			515 517-518 520-522 524 526 529 531-532
			537 539 542 545-546 551 556 571 577 580
			589-590 592 596 601 622 631-632 640 642
			654-655 664 666 668-669 673 684 703 708
			716 718 721 723 727 733 735 743-744 754
			758 765 771 775 777-779 783-786 797-799
			810 816-817 828 834 845 859 861 863 870
			878-879 881 884 887 890 897 904 907 910
			912-913 918 923 929 939-942 945-947
leukocyte	GIBCO	LUC001	2-4 7-8 13-17 20-23 31-33 38 43 48-49 51
			53-57 63 66-68 74-78 85-88 93 97 122 124
			129 131-132 134 137 140 163 166 168 171-
			172 175 188-192 197 200 208-213 216 221
			223 231 236 242 252 257-258 261-263 268-
			270 272 277 279 287-288 294 307 311 314
			316-326 329 337-339 341 373-374 376-377
			381 388-392 396 400-401 413-414 423 436
			441 450 454-455 459 463 465 467 489 491-
			493 495 498-499 504-505 507 514 518 520-
			522 524 526 529 531 533 536-537 539-540
			545 552-554 556 568 571 577 580 585 589-
			590 596 599 602 605 607 610 612-613 615
			618-619 621-622 625 638 640 642-644 664
			667 677-678 684 690-693 696 700 703-704
			707 713 715-718 721 727 734-735 738 740-
			746 748 753-754 758 775 778 780 789 794
			797-798 801-802 815 817 825 827 829 834
			836 846-847 850 859-861 863-864 866-867
			871-872 878 884 886-888 891 896-897 902
			904 910 913 916 921 923-926 929-932 936
			943
leukocyte	Clontech	LUC003	12 14-15 18 32 111 134 137 172 221 277 280
			311 316-325 436 454 467 549 552 568 585
			603 643 691-692 698-699 734 744 751 754
			784 797-798 861 897 916 923
Melanoma from	Clontech	MEL004	3-4 16 20-21 43 46 48 97 103 147 163 188
cell line ATCC			191 213 216 221 231 241 245-246 260 262-
CRL #1424			263 316-325 381 407 431 504 525 527 542
			556 568 577 589 596 607 613 676 693 714
			735 737-739 744 758-760 775 822 850 863
			878 887 897
mammary gland	Invitrogen	MMG001	1-8 14-18 20-21 25 28-29 37 39 43 49 51-57
			60 66-67 72 75-76 87 95 97 103-104 106 112
			115 119 122 127 130 134 137 139 142 150
			166-168 172 175 184 186 188-189 191-192
			200 213-214 222-224 226-229 240 252 257-
			259 263 267 271 276 278 282 287-288 299
			301-302 305 307-308 311-313 316-325 327-
			328 332 340-341 358 360-362 369 373 378
			381 383 388 390-392 397 402-403 409 415-
			416 423 425 428-429 433 436 444 454-456
			459-460 464 467 469 481 483-484 486 493
			495 498-499 515 524-525 529-530 532-537
			541-542 545 551-552 562 582-583 586-587
			593 599-600 602 604-605 610 618-619 622
			625-627 634 644 646-647 652 654-655 662
			673 676 680 684 687-688 691-692 701-703
			715 717 721 723 726 735 743 751 754 758
			765-766 771 777-778 789 803 805 807 809-
			811 821 827 829 850 860 887-888 892 896
			898 901-902 905 911 913 917 925-926 930
			936 939-942
induced neuron	Stratagene	NTD001	2 16 32 51 66 88 97 124 130 134 137 172
cells			188-189 191-192 231 252 257 260 277 291
			373 424 431 454 460 489 495 523 525 582-
			583 591 631-632 643 649 670 695 725-726
			735 765 789 797-798 837 850 878 884 888
			890 913 929 946
retinoid acid	Stratagene	NTR001	2 5-6 20-22 136-137 188-194 197-198 224
induced			311 375 381 410 457 462 475 495 531 546
neuronal cells			548 552 599 618 678 743 752 819 828 890
			895 897 930 934 938 944 946
neuronal cells	Stratagene	NTU001	2 5-6 20-21 55-56 87 137 188-192 197-198
			215-216 260 287 291 310-311 316-325 365
			375 423 457 459 470 499 532 542 564 576
			598-599 623 643 651-652 673 721 726 743
			745 752 754 765 780 787 789 822 829 870
			875 888 896 917 919 929
pituitary gland	Clontech	PIT004	41-42 83 85 97 134 193-194 204 208 213 224
			257-258 263-265 285-286 308 311 360-361
			413 443 445 491 514 529 532 639 644 647
			682 701-702 716 781 822 829 836 850 933
			939-941 947
placenta	Clontech	PLA003	16 31 34 49 66 80 87 97 101-102 134 158
			165 172 179 184 188 197-198 209-210 218
			220 229 235 249 256 267-270 277 287-288
			302 307 332 360-361 365 388 394 414 441
			444 454 457 460 493 498-500 505-506 509
			529 531-532 550 559-560 564 572 587 601
			625 630-632 638 672 682-684 689 706 708
			726 733 735 744 754 761 784-786 793 863
			875 897 924 929 937
prostate	Clontech	PRT001	7-8 51 85 87 97 100 122 134 139 214 216
			221 231 257 271 276 335 337-338 392 400-
			401 431 440 459 477 530 534-535 546 556
			582-583 599 622 631-632 639 651 663-664
			673 683 707 715 735 740 765 773-774 777
			810 823 897 909 919 934 939-941 947
rectum	Invitrogen	REC001	18 54 66 134 137 169 188 200 213 225 251
			263 288 311-313 316-325 340 388 423 429
			441 454 459 514 532 542 610 626 646 651
			657 715 719 723 728 735 740 758 766 785-
			786 823 829 833 836 886 942
salivary gland	Clontech	SAL001	31 49 78 95 134 136-137 143 176 188 208
			223 244 268-270 284 308 311 316-325 388-
			389 391 436 441 459 476 514-515 520-521
			532 543 568 589 596 610 619 684 691-692
			713 718 727 736 754 777 824 836 864 867
			878 883 897 901-902 916-917 933 938-941
salivary gland	Clontech	SALs03	460
skin fibroblast	ATCC	SFB001	379-380 850
skin fibroblast	ATCC	SFB002	742 850
skin fibroblast	ATCC	SFB003	87
small intestine	Clontech	SIN001	27-29 31 38 40 46 48 51 54 57-58 62 65 67
			75 77 85 97 110 112 116-117 119 131-132
			134 137 140 161 163 166 168 177 188 197-
			198 208 213 220 224 229 246 257 261-262
			264-265 276-277 288 295 297 299 311 316-
			326 328-330 337-338 340 360-361 373 375
			382 390-391 410 413 428-429 436 438 440
			453-454 459 468 476-477 497 507 511 522
			531 536 538 542 545-546 548 552 556 564
			570-571 576 580-581 586-587 591 596 599
			605 610 613 619 625-626 643-644 651-653
			664-666 668-670 677 680 684 693 700-702
			706-707 713-715 723-724 729-730 735 740
			746 748 753-754 757-758 764 777-778 784-
			786 818 822 824 826-829 833-837 842 862-
			863 865-867 877-878 886 897 900-902 906
			913 916 921 925-926 936 939-941
skeletal muscle	Clontech	SKM001	42 98 156 163 191-192 200 261 305 311 395
			415 462 468 504 531 543 566 582-583 585
			594 680 740 853 875 927 933 935
skeletal muscle	Clontech	SKM002	850
spinal cord	Clontech	SPC001	18 23 33 37 42 51 67 87 92 94 97 100 140
			162 184 188 191-192 208 213 220 231 248
			262 268-271 273-274 282 287-288 290 307
			311 316-325 358 364 376-377 383 387 389-
			390 402 412 422 444 455 476 483-484 489
			504 522 534-535 556 562 587-588 591 597
			603-604 618-619 643 651 667-670 677 693
			703-704 717-718 727 746 757 773 808 810
			827 834-835 837 850 871-872 875 904 910
			931-932 939-941
adult spleen	Clontech	SPLc01	33 38 57 67 75 87 134 142 163 216 221 229
			244 257 304 307 311 316-325 340 355-356
			378 441 468 525 538 545 560 564 599 721
			754 766 780 794 827 841 850 866
stomach	Clontech	STO001	18 65 88 163 188 208 213 261 272 277 286
			294 336 373 396 412 459 514 553 602 610
			647 651-652 671 673 714 774 790 831 833
			842 850 876
thalamus	Clontech	THA002	2 87 96 103 106 189-192 208 252 258 295
			308 311 367 376-377 383-384 445 455 459-
			460 498 529 587 598 602 629 654-655 705-
			706 715 717 723 754 775 810 817 822 864
			867 881 892 927 930
thymus	Clontech	THM001	3-4 8 18 28 54 57 63 65 68 84 97 100 116-
			117 122 134 142 151 169 171-172 188 195
			197-198 201 213 221 237 245 261 287 311
			316-325 360-361 376-377 423 441 444 459
			489 491-493 495 498 504 507 514 527 532
			534-536 539 553 556 568 571-572 590 595-
			596 599 610 618 622 631-632 643 647 651
			654-655 664 687-688 691-693 703 715 721
			733-735 748 760 762 765 781 794 799 802
			831 834 836 842 850 860-861 863 871 878
			885 896-897 903 910 923 925-926 928 939-
			941
thymus	Clontech	THMc02	2-4 17 20-22 37-38 42-43 46 63 65-68 76 88
			95 103 118 120 124 134 137 140-141 143 163
			165 171 179 182 189-194 198 200 212-215
			221 226-228 231 244 257 262 266 276-277
			287-288 297 299 307 316-325 341 352 358
			360-361 373 376-377 379-381 389 391 394-
			396 403 410-411 436 440 445 450 459 463-
			464 469 478 491 495 500 507 511 519-521
			530 532 539 542 550 555 560 563 576 581
			587 595 601 610-611 613-614 618 622 625-
			626 631-632 638 642-644 657 664 667 670
			673 680 683 687 691-693 699 715-716 721
			740 743-744 747-754 761 763-765 771 777
			780-781 784-787 790 794 805 811 820 826
			831 834 841 845 861 867-868 878 881 883
			891 893-894 896-897 902 904 910 912-914
			918 923 936-941 946-947
thyroid gland	Clontech	THR001	1-2 18-21 27 32 38 42 46 49 51 53-56 66 72
			77-78 87-88 97 115 119 124 130-131 134 136
			152 163-165 172 183 188-192 202 212-213
			216 221 224 229 235 241-243 252 257-258
			261 263-265 267 277 279 297 301 305 308
			311 316-325 327 357 363 373 376-377 381
			383 389 397 400-401 410 413-414 427-428
			443-444 446-447 457 459 463 467-469 475
			482 489 495 499-500 504 509 513 519-522
			526 529 533 537-538 542 545-546 548 556
			564 567-568 582-583 589 592 599 605 608
			611 621 623 630 642-644 648 651-652 654-
			655 664 672-676 684-686 691-694 700 706-
			708 713 717-718 721-722 725 729 731 734-
			735 740 748 753-754 760 764 766 771 774
			777 781 792 797-800 802 805 826 828-829
			834 842 850 861 863 868 876 879 897 899
			901 910 913 929 937 939-941
trachea	Clontech	TRC001	20-21 38 112 161 163 188 263 267 327 413
			420 457 459-460 471 514 540-541 552 572
			574 622 639 654-655 676-677 691-692 707
			725 743 748 765 777-778 862 868 897 905
			908 944
uterus	Clontech	UTR001	51 67 126 130 133 140 188-192 229 267 329
			373 440 491 514 599 685-686 693 713 716-
			717 735 897 905 911 939-941
*The 16 tissue-mRNAs and their vendor source, are as follows: 1) Normal adult brain mRNA (Invitrogen), 2) normal adult kidney mRNA (Invitrogen), 3) normal adult liver mRNA (Invitrogen), 4) normal fetal brain mRNA (Invitrogen), 5) normal fetal kidney mRNA (Invitrogen), 6) normal fetal liver mRNA (Invitrogen), 7) normal fetal skin mRNA (Invitrogen), 8) human adrenal gland mRNA (Clontech), 9) human bone marrow mRNA (Clontech), 10)
# human leukemia lymphablastic mRNA (Clontech), 11) human thymus mRNA (Clontech), 12) human lymph node mRNA (Clontech), 13) human spinal cord mRNA (Clontech), 14) human thyroid mRNA (Clontech), 15) human esophagus mRNA (BioChain), 16) human conceptional umbilical cord mRNA (BioChain).

TABLE 2
			SMITH-
SEQ	ACCESSION		WATERMAN	%
ID NO:	NUMBER	DESCRIPTION	SCORE	IDENTITY
1	Z99162	Schizosaccharomyces pombe putative transporter	134	30
2	U44839	Homo sapiens UHX1 protein	3719	100
3	AF031939	Mus musculus RalBP1-associated EH domain protein Reps1	3687	94
4	AF031939	Mus musculus RalBP1-associated EH domain protein Reps1	1887	57
5	U69490	Mus musculus p56lck-associated adapter protein Lad	173	37
6	U69490	Mus musculus p56lck-associated adapter protein Lad	173	37
7	AL161538	Arabidopsis thaliana disease resistance N like protein	398	33
8	W88660	Secreted protein encoded by gene 127 clone HSUBW09.	175	97
9	D50617	Saccharomyces cerevisiae YFL025C	306	34
10	AB028070	Homo sapiens activator of S phase Kinase	193	42
11	Y27676	Human secreted protein encoded by gene No. 110.	474	100
12	Y30721	Amino acid sequence of a human secreted protein.	355	98
13	AF257330	Homo sapiens COBW-like protein	566	97
14	AF089812	Mus musculus ubiquitin-conjugating enzyme HR6A	378	98
15	AF089812	Mus musculus ubiquitin-conjugating enzyme HR6A	688	90
16	Y94959	Human secreted protein clone mc300_1 protein sequence SEO ID NO: 124.	204	97
17	AF212247	Homo sapiens CDA08	2664	92
18	AF064868	Rattus norvegicus brain-enriched guanylate kinase-associated	452	45
		protein 1; BEGA1
19	W67863	Human secreted protein encoded by gene 57 clone HFEBF41.	551	98
20	AL132954	Arabidopsis thaliana putative protein	201	25
21	AL132954	Arabidopsis thaliana putative protein	361	34
22	Y48359	Human prostate cancer-associated protein 56.	403	98
23	AF202892	Mus musculus Kif21a	5679	92
24	W75143	Human secreted protein encoded by gene 23 clone HBMCT32.	148	100
25	Y35921	Extended human secreted protein sequence, SEQ ID NO. 170.	548	99
26	Y27587	Human secreted protein encoded by gene No. 21.	448	100
27	AF190900	Homo sapiens kelch-like protein C3IP1	1767	74
28	X12517	Homo sapiens C protein (AA 1-159)	903	99
29	AY014403	Homo sapiens kinesin-like protein RBKIN1	9290	99
30	AF240783	Mus musculus ELKL motif kinase 2 short form	194	48
31	Y02661	Human secreted protein encoded by gene 12 clone HFTCU19.	858	81
32	AF271070	Homo sapiens amino acid transporter system A1	2466	100
33	W75151	Human secreted protein encoded by gene 34 clone HTEGA81.	507	100
34	AF206329	Mus musculus polydom protein	3886	81
35	AF162224	Mus musculus angiopoietin-related protein 3	138	23
36	Y35996	Extended human secreted protein sequence, SEQ ID NO. 361.	503	97
37	AF022891	Drosophila melanogaster Fuzzy	248	30
38	X76775	Homo sapiens HLA-DMA	1091	100
39	Y16045	Arabidopsis thaliana leucine-rich repeat protein	160	33
40	AC003077	Homo sapiens 60% similar to AB002297 (PID:g2224539)	2629	61
41	W74887	Human secreted protein encoded by gene 160 clone HCELB21.	203	100
42	AC005390	Homo sapiens R31180_1	1550	66
43	M98450	Oryctolagus cuniculus casein kinase-II beta	668	100
44	Y10840	Amino acid sequence of a human secreted protein.	349	100
45	AL050231	Drosophila melanogaster alternatively spliced form	203	46
46	U58280	Mus musculus second largest subunit of RNA polymerase I	4956	89
47	AJ224819	Homo sapiens tumor suppressor	457	42
48	AF205935	Mus musculus MGA protein	3477	77
49	AF095352	Homo sapiens RAB-like protein 2B	718	99
50	L20321	Homo sapiens protein serine/threonine kinase	394	41
51	Y13374	Homo sapiens putative prenylated protein	397	87
52	AJ302031	Rattus norvegicus putative alpha 1B-glycoprotein	519	40
53	AF145681	Drosophila melanogaster BcDNA.LD23181	828	47
54	AJ132192	Mus musculus HS1 binding protein 3	1454	75
55	AF145681	Drosophila melanogaster BcDNA.LD23181	692	53
56	AB036800	Drosophila melanogaster egg-derived tyrosine phosphatase	828	47
57	Y87327	Human signal peptide containing protein HSPP-104 SEQ ID NO: 104.	584	100
58	Y36237	Human secreted protein encoded by gene 14.	177	100
59	Y87310	Human signal peptide containing protein HSPP-87 SEQ ID NO: 87.	370	100
60	AF062476	Mus musculus retinoic acid-responsive protein; STRA6	1437	74
61	Y38394	Human secreted protein encoded by gene No. 9.	213	100
62	AL139421	Homo sapiens dJ717I23.1	3267	100
		(novel protein similar to Xenopus laevis Sojo protein)
63	AL359782	Trypanosoma brucei possible (hhv-6) u1102, variant a dna,	142	50
		complete virion genome.
64	Y19561	Amino acid sequence of a human secreted protein.	514	100
65	L12690	Homo sapiens neutrophil peptide-1	493	100
66	AF274057	Rattus norvegicus GRIP-asaociated protein 1 long form	3814	92
67	AL162458	Homo sapiens bA465L10.2	6467	99
		(novel C2H2 type zinc finger protein similar to chicken FZF-1)
68	Y87100	Human secreted protein sequence SEQ ID NO: 139.	267	100
69	Y86320	Human secreted protein HPRBC80, SEQ ID NO: 235.	361	100
70	Z99162	Schizosaccharomyces pombe putative transporter	169	27
71	S67057	Cricetulus migratorius = Armenian hamsters, Peptide,	158	71
		223 aa serum amyloid P, SAP, female protein, FP = pentraxin
72	Y36183	Human secreted protein #55.	449	96
73	U08813	Oryctolagus cuniculus 597 aa protein related to Na/glucose cotransporters	1231	85
74	AB009883	Nicotiana tabacum KED	202	23
75	AX015323	Homo sapiens hFATP1	3367	99
76	AB013361	Homo sapiens DPM2	153	100
77	Y25732	Human secreted protein encoded from gene 22.	212	100
78	M37033	Homo sapiens CD53 glycoprotein	116	95
79	Y36098	Extended human secreted protein sequence, SEQ ID NO. 483.	193	100
80	AB021644	Homo sapiens gonadotropin inducible transcription repressor-4	886	60
81	U68267	Mus musculus myosin binding protein H	1202	66
82	Y36332	Human secreted protein encoded by gene 109.	268	100
83	AF254956	Homo sapiens candidate tumor suppressor protein	2030	99
84	L12690	Homo sapiens neutrophil peptide-1	493	100
85	AF295378	Homo sapiens MAGEF1	754	50
86	AF144627	Mus musculus SLIT1	283	32
87	AF208536	Homo sapiens nucleotide binding protein; NBP	1372	100
88	Y59657	Secreted protein 108-003-5-0-A8-FL.	689	100
89	Y27626	Human secreted protein encoded by gene No. 60.	352	100
90	AF035268	Homo sapiens phosphatidylserine-specific phospholipase A1	498	41
91	Y12861	Human 5′ EST secreted protein SEQ ID NO: 451.	448	100
92	Y53049	Human secreted protein clone cj378_3 protein sequence SEQ ID NO: 104.	463	100
93	Y41354	Human secreted protein encoded by gene 47 clone HUFCJ30.	288	100
94	W74777	Human secreted protein encoded by gene 48 clone HFCAI74.	245	100
95	AB040610	Homo sapiens glycoprotein beta-Gal 3′-sulfotransferase	388	47
96	D32215	Danio rerio emx2 homeoprotein	1242	92
97	Y94959	Human secreted protein clone mc300_1 protein sequence SEQ ID NO: 124.	204	97
98	AF024496	Caenorhabditis elegans contains similarity to Plasmodium falciparum	390	30
		glycophorin-binding protein homolog 2 (GB:X69769)
99	A09779	Homo sapiens interferon-gamma receptor segment binding interferon-gamma	600	99
100	AE002030	Deinococcus radiodurans thermoresistant gluconokinase	246	52
101	AB052620	Mus musculus DDM36	4361	89
102	M69245	Homo sapiens pregnancy-specific beta-1-glycoprotein	613	70
103	X92841	Homo sapiens MHC class I chain-related protein A	588	100
104	AF136401	Rattus norvegicus TRP2	410	88
105	AL031709	Homo sapiens c316G12.4	561	93
		(novel protein similar to API1 and API2
		(apoptosis inhibitor 1 and 2 (MIHB, MIHC, IAP1, IAP2)))
106	Y38389	Human secreted protein encoded by gene No. 4.	152	90
107	Y27582	Human secreted protein encoded by gene No. 16.	320	100
108	AF130079	Homo sapiens PRO2852	231	60
109	J02818	Oryctolagus cuniculus cytochrome P-450p-2	893	46
110	Y07894	Human secreted protein fragment encoded from gene 43.	169	48
111	AF119297	Homo sapiens neuroendocrine-specific protein-like protein 1	240	97
112	X56203	Plasmodium falciparum liver stage antigen	254	23
113	Y76200	Human secreted protein encoded by gene 77.	262	100
114	Y36270	Human secreted protein encoded by gene 47.	359	100
115	AJ133120	Rattus norvegicus Proline rich synapse associated protein 2	3938	93
116	AL031709	Homo sapiens c316G12.4	299	84
		(novel protein similar to API1 and API2
		(apoptosis inhibitor 1 and 2 (MIHB, MIHC, IAP1, IAP2)))
117	AL031709	Homo sapiens c316G12.4	582	100
		(novel protein similar to API1 and AP12
		(apoptosis inhibitor 1 and 2 (MIHB, MIHC, IAP1, IAP2)))
118	AB018542	Homo sapiens CD98 light chain	1829	86
119	AB012692	Homo sapiens CAC-1	132	41
120	AB021644	Homo sapiens gonadotropin inducible transcription repressor-4	2050	57
121	AF110776	Homo sapiens adrenal gland protein AD-003	542	53
122	AL391688	Homo sapiens bA524D16A.2.1	683	42
		(novel protein similar to mouse granuphilin-a)
123	AB042624	Homo sapiens SIRP-b2	247	50
124	AF019980	Dictyostelium discoideum ZipA	280	22
125	U58917	Homo sapiens IL-17 receptor	266	30
126	AK021852	Homo sapiens unnamed protein product	884	99
127	AY007378	Homo sapiens G-protein beta subunit-like protein	1731	99
129	AF195522	Trypanosoma cruzi B-cell mitogen precursor	345	39
130	Z81587	Caenorhabditis elegans contains similarity to Pfam domain: PF01363	276	42
		(FYVE zinc finger), Score = 65.2, E-value = 1.9e-17, N = 1
131	AF188700	Homo sapiens actin filament associated protein	844	43
132	M77678	Mus musculus NKR-P1 gene-40 protein	118	26
133	AF217226	Homo sapiens zinc finger protein ZNF286	574	94
134	L06505	Homo sapiens ribosomal protein L12	841	99
135	Y12902	Human 5′ EST secreted protein SEQ ID NO: 492.	134	100
136	L11672	Homo sapiens zinc finger protein	3059	60
137	U47924	Homo sapiens B-cell receptor associated protein	925	100
138	AC006271	Homo sapiens BC319430_7	163	49
139	AE000799	Methanothermobacter thermoautotrophicus O-linked GlcNAc transferase	151	37
141	Y13037	Human secreted protein encoded by 5′ EST SEQ ID NO: 51.	187	100
142	AJ3271735	Homo sapiens sprouty (Drosophila) homolog 3	170	32
143	AF228917	Rattus norvegicus small rec	454	56
144	U41552	Caenorhabditis elegans Contains similarity to Pfam domain: PF00122	127	50
		(E1-E2_ATPase), Score = 36.1, E-value = 8.1e-10, N = 4
145	AL049569	Homo sapiens dJ37C10.3 (novel ATPase)	328	45
146	AF109888	Macaca mulatta sodium-calcium exchanger circular exon 2 transcript	147	32
147	AF217227	Homo sapiens zinc finger protein ZNF287	1146	43
148	B24426	Human PR01286 protein sequence SEQ ID NO: 199.	466	100
149	AB019120	Rattus norvegicus seven transmembrane receptor	1015	37
150	AF062476	Mus musculus retinoic acid-responsive protein; STRA6	190	72
151	X64223	Mus musculus Fc-E receptor II (Fc-ERII/CD23)	138	34
152	X83543	Homo sapiens APXL	412	41
153	AF217289	Homo sapiens cadherin 20	4170	99
154	Y36310	Human secreted protein encoded by gene 87.	250	100
155	D50577	Mesocricetus auratus carboxylesterase precursor	441	50
156	AB027004	Homo sapiens protein phosphatase	229	41
157	AF305071	Mus musculus calsenilin-like protein	1281	99
158	Y13126	Human secreted protein encoded by 5′ EST SEQ ID NO: 140.	160	96
159	AB016215	Cyprinus carpio complement C3-Q2	132	35
160	AF294278	Homo sapiens PR-domain-containing protein 16	6646	99
161	AF305210	Homo sapiens concentrative Na+− nucleoside cotransporter hCNT3	3609	100
163	U10281	Sus scrofa gastric mucin	165	22
164	M13095	Rattus sp. 0-44 protein	661	98
165	AY008763	Homo sapiens sentrin/SUMO-specific protease	537	53
166	S72304	Mus ap. LMW G-protein	763	94
167	AF238315	Homo sapiens HZFw1 protein	2251	99
168	M22414	Homo sapiens ribonuclease inhibitor precursor	222	30
169	AL035702	Homo sapiens dJ593C16.1	3010	58
		(ras GTPase activating protein)
170	AL110500	Caenorhabditis elegans Y87G2A.13	146	22
171	AB028860	Mus musculus mDj10	189	37
172	M83653	Homo sapiens cytoplasmic phosphotyrosyl protein phosphatase	648	100
173	AB041601	Mus musculus unnamed protein product	255	61
174	M35012	Drosophila melanogaster non-muscle myosin heavy chain	200	24
175	D88577	Mus musculus Kupffer cell receptor	904	46
176	Y30847	Human secreted protein encoded from gene 37.	239	100
177	Y01390	Secreted protein encoded by gene 8 clone HTXDJ88.	301	100
178	M23725	Homo sapiens M2-type pyruvate kinase	152	78
179	X72875	Homo sapiens complement factor B	3527	100
180	Y08420	Homo sapiens nicotinic acetylcholine receptor alpha7 subunit precursor	1794	100
181	M24766	Homo sapiens alpha-2 type IV collagen	3756	99
183	AL110500	Caenorhabditis elegans Y87G2A.13	180	25
184	AF286598	Homo sapiens angiostatin binding protein 1	1447	62
185	Y13084	Human secreted protein encoded by 5′ EST SEQ ID NO: 98.	130	62
186	AF151067	Homo sapiens HSPC233	892	58
187	AL133283	Homo sapiens bA31M2.1	1629	100
		(novel protein similar to the GLI family of zinc finger proteins)
188	J03799	Homo sapiens laminin-binding protein	1331	94
189	M27132	Homo sapiens ATP synthase beta subunit precursor	734	98
190	M27132	Homo sapiens ATP synthase beta subunit precursor	206	100
191	M27132	Homo sapiens ATP synthase beta subunit precursor	709	98
192	M27132	Homo sapiens ATP synthase beta subunit precursor	1127	89
193	U76638	Homo sapiens BRCA1-associated RING domain protein	4101	100
194	U76638	Homo sapiens BRCA1-associated RING domain protein	394	100
195	S57688	Thermotoga maritima EF-G	289	75
196	U09453	Cricetulus griseus UDP-N-acetylglucosamine:	239	85
		dolichyl phosphate N-acetylglucosamine 1-phosphate transferase
197	D86821	Streptomyces coelicolor PrfB	151	40
198	Y36068	Extended human secreted protein sequence, SEQ ID NO. 453.	495	94
199	U38980	Homo sapiens hPMSR6	408	92
200	Y08999	Homo sapiens Sop2p-like protein	572	99
201	AJ401272	Canis familiaris Band4.1-like5 protein	2233	95
202	U09608	Homo sapiens cell surface protein	292	84
203	Y64747	Human 5′ EST related polypeptide SEQ ID NO: 908.	471	96
204	Y66754	Membrane-bound protein PR01187.	656	100
205	U52111	Homo sapiens Ca2+/Calmodulin-dependent protein kinase I	1680	99
206	AF279265	Homo sapiens putative anion transporter 1	186	39
207	X91911	Homo sapiens rtvp-1	446	40
208	X56390	Canis familiaris rac2	508	100
209	AK026888	Homo sapiens unnamed protein product	1536	100
210	AK026888	Homo sapiens unnamed protein product	927	98
211	AB044805	Homo sapiens 6-phosphofructo-2-kinase heart isoform	2452	100
212	AB013897	Homo sapiens HKR1	3083	99
213	U37351	Mus musculus Paneth cell enhanced expression PCEE	746	89
214	AJ278475	Homo sapiens transport-secretion protein 2.1 (TTS-2.1)	2179	98
215	U58749	Caenorhabditis elegans coded for by C. elegans cDNA yk8c7.5;	844	63
		coded for by C. elegans cDNA yk47c5.5;
		coded for by C. elegans cDNA yk76b5.5;
		coded for by C. elegans CDNA yk8c7.3;
		coded for by C. elegans CDNA yk47c5.3;
		strong similarity to catalytic domains of ser/thr protein kinases
216	AF000195	Caenorhabditis elegans Contains similarity to Pfam domain:	1072	47
		PF00169 (PH), Score = 20.6, E-value = 1.9e-05, N = 1
217	AB016768	Mus musculus thrombospondin type 1 domain	189	40
218	AB026256	Homo sapiens organic anion transporter OATP-B	1928	99
219	X56692	Homo sapiens C-reactive protein	327	100
220	AF064801	Homo sapiens multiple membrane spanning receptor TRC8	603	29
221	AF191545	Homo sapiens aminopeptidase	5048	99
222	X66171	Homo sapiens CMRF-35 antigen	228	33
223	Z38061	Saccharomyces cerevisiae mal5, stal, len: 1367, CAI: 0.3,	241	22
		AMYH_YEAST P08640 GLUCOAMYLASE S1 (EC 3.2.1.3)
224	AC005306	Homo sapiens R27216_1	1560	100
225	AF176532	Mus musculus F-box protein FBX17	1059	81
226	AF097432	Homo sapiens GROS1-L protein	2147	99
227	AF097432	Homo sapiens GROS1-L protein	1900	96
226	AF097432	Homo sapiens GROS1-L protein	3881	99
229	Z29094	Caenorhabditis elegans contains similarity to Pfam domain:	388	32
		PF01699 (Sodium/calcium exchanger protein),
		Score = 268.7, E-value = 2.5e-77, N = 2
230	Y66669	Membrane-bound protein PRO839.	366	100
231	X75931	Bos taurus Cleavage and Polyadenylation	4033	98
		specificity factor (CPSF) 100 kD subunit
232	AF152562	Homo sapiens angiopoietin-related protein 3	1210	99
233	L19686	Homo sapiens macrophage migration inhibitory factor	564	94
234	AL008723	Homo sapiens dj90G24.4 (SAAT1	3408	100
		(low affinity sodium glucose cotransporter
		(sodium:solute symporter family)))
235	U50927	Rattus norvegicus zinc transporter ZnT-2	1040	85
236	AF099973	Mus musculus schlafen2	982	53
237	U67557	Methanococcus jannaschii cell division control protein 48 (cdc48),	1050	41
		AAA family
238	AF207661	Homo sapiens sodium bicarbonate cotransporter-like protein	5645	100
239	AF284337	Homo sapiens SEBOX	1209	100
240	AF081669	synthetic construct VU91B calmodulin	109	42
241	U96166	Streptococcus cristatus srpA	286	18
242	AE005024	Halobacterium sp. NRC-1 Vng0821c	130	37
243	AK023335	Homo sapiens unnamed protein product	2344	99
244	AK024464	Homo sapiens FLJ00057 protein	3033	99
245	U55376	Caenorhabditis elegans coded for by C. elegans CDNA cm21e6;	752	40
		coded for by C. elegans CDNA cm01e2;
		similar to melibiose carrier protein (thiomethylgalactoeide permease II)
246	AK022660	Homo sapiens unnamed protein product	1173	99
247	D16235	Bos taurus PLC alpha	156	24
248	B08894	Human secreted protein sequence encoded by gene 4 SEQ ID NO: 51.	235	67
249	AJ289709	HERV-H/env62 envelope protein	754	39
250	AJ242540	Volvox carteri f. nagariensis hydroxyproline-rich glycoprotein DZ-HRGP	209	63
251	Y66747	Membrane-bound protein PRO1158.	609	100
252	AL031532	Schizosaccharomyces pombe ubiquitin conjugating enzyme	236	41
253	W74899	Human secreted protein encoded by gene 172 clone HODCW06.	197	100
254	D43633	Oryzias latipes G protein-coupled seven-transmembrane receptor	462	36
255	D43633	Oryzias latipes G protein-coupled seven-transmembrane receptor	823	42
256	AJ277750	Homo sapiens UBASH3A protein	1035	44
257	Y12711	Homo sapiens putative progesterone binding protein	187	42
258	W74939	Human secreted protein encoded by gene 49 clone HAGBI17.	211	100
259	D16432	Mus musculus murine homologue of CD63/ME491	504	99
260	AB031051	Homo sapiens organic anion transporter OATP-E	863	37
261	J03998	Plasmodium falciparum glutamic acid-rich protein	139	29
262	U80953	Caenorhabditis elegans weakly similar in serine repeat region to rat	336	32
		thyroxine-binding globulin (PIR:A39567)
		and to D. melanogaster ecdysone-inducible protein E75-C
		(SP:E75C_DROME, P13055)
263	AF155662	Homo sapiens putative 16.7 kDa protein	766	99
264	Z46237	Saccharomyces cerevisiae putative protein	245	27
265	Z46237	Saccharomyces cerevisiae putative protein	245	27
266	AF208795	Ictalurus punctatus NCC receptor protein 1	286	50
267	AJ249901	Mus musculus secreted modular calcium-binding protein 2	2332	95
268	X05908	Homo sapiens lipocortin (AA 1-346)	967	100
269	X05908	Homo sapiens lipocortin (AA 1-346)	1511	100
270	X05908	Homo sapiens lipocortin (AA 1-346)	967	100
271	AC003038	Homo sapiens R30923_1	2992	100
273	U37143	Homo sapiens cytochrome P450 monooxygenase CYP2J2	942	44
274	U37143	Homo sapiens cytochrome P450 monooxygenase CYP2J2	555	45
275	AF154933	Sus scrofa complement component C3	374	39
276	Z54342	Caenorhabditis elegans contains similarity to Pfam domain:	570	39
		PF00328 (Histidine acid pliosphatase), Score = 511.8,
		E-value = 2e-152, N = 1
277	M20259	Homo sapiens thymosin beta-10	169	97
278	Y91386	Human secreted protein sequence encoded by gene 41 SEQ ID NO: 107.	558	100
279	AF151110	Mus musculus COP1 protein	2268	99
280	AF117959	Homo sapiens CDK4-binding protein p34SEI1	114	68
281	Y91370	Human secreted protein sequence encoded by gene 25 SEQ ID NO: 91.	293	100
282	AF079446	Dictyostelium discoideum developmental protein DG1067	520	38
283	M60618	Homo sapiens nuclear autoantigen	300	84
284	D83777	Homo sapiens expressed ubiquitously with strong expression in brain	912	51
285	L77864	Homo sapiens stat-like protein	290	96
286	Y02775	Human secreted protein encoded by gene 12 clone HFTCU19.	288	98
287	L21998	Homo sapiens mucin	389	24
288	AF184275	Mus musculus F-box protein FBX18	4409	92
289	AF053356	Homo sapiens leucin rich neuronal protein	264	37
290	AC016829	Arabidopsis thaliana putative O-linked GlcNAC transferase	377	27
291	Y73414	Human secreted protein clone yb101_1 protein sequence SEQ ID NO: 50.	472	100
292	W88615	Secreted protein encoded by gene 82 clone HNGBT31.	525	97
293	AJ278018	Homo sapiens calsyntenin-2	5080	100
294	Y41460	Fragment of human secreted protein encoded by gene 42.	1034	100
295	D79995	Homo sapiens similar to pig tubulin-tyrosine ligase.	415	42
296	AF092878	Homo sapiens zinc RING finger protein SAG	323	92
297	AL137784	Homo sapiens dJ199J3.1	561	100
		(novel protein similar to ubiquitin carboxyl - terminal hydrolase 16
		(EC 3.1.2.15))
298	Z70310	Caenorhabditis elegans contains similarity to Pfam domain:	520	37
		PF00013 (KH domain), Score = 42.8, E-value = 3 .7e-12, N = 1;
		PF00023 (Ank repeat), Score = 428.2, E-value = 2.4e-125, N = 19
299	Y48600	Human breast tumour-associated protein 61.	288	98
300	AB007889	Homo sapiens KIAA0429	386	46
301	U64601	Caenorhabditis elegans Gene probably begins in the next cosmid	412	50
302	U27109	Homo sapiens prepromultimerin	208	24
304	AF278532	Homo sapiens beta-netrin	3347	99
305	X51957	Homo sapiens muscle-specific enolase	555	94
306	Y07895	Human secreted protein fragment encoded from gene 44.	537	100
307	AE001045	Archaeoglobus fulgidus proliferating-cell nucleolar antigen P120, putative	144	41
308	AB033882	Coturnix japonica protein kinase C inhibitor	433	60
309	AC006284	Arabidopsis thaliana putative ankyrin	198	44
310	AF093673	Cricetulus griseus layilin	576	45
311	J02642	Homo sapiens glyceraldehyde 3-phosphate dehydrogenase (EC 1.2.1.12)	1685	97
312	AB012692	Homo sapiens CAC-1	560	94
313	AB012692	Homo sapiens CAC-1	484	73
314	Y17571	Homo sapiens aralar2	300	48
315	X15324	Homo sapiens angiotensinogen	1925	97
316	X97321	Homo sapiens HLA-C protein	1099	98
317	M24036	Homo sapiens MHC HLA-B8 chain	1033	92
318	U04245	Homo sapiens MHC class I antigen	940	95
319	M32318	Homo sapiens HLA protein allele B57	982	88
320	U41057	Homo sapiens HLA class I A locus antigen A*68new	1801	93
321	M32321	Homo sapiens HLA protein allele A25	1163	96
322	AJ250917	Homo sapiens human leucocyte antigen B	315	88
323	D64147	Homo sapiens HLA-Cw*0602	1121	96
324	AB005048	Homo sapiens A26null allele	977	85
325	M24043	Homo sapiens MHC HLA-A1 chain	1123	97
326	AF190900	Homo sapiens kelch-like protein C3IP1	2982	100
327	AB022023	Bos taurus nonmuscle myosin heavy chain B	158	22
328	X85019	Homo sapiens UDP-GalNAC:polypeptide N-acetylgalactosaminyl	1431	50
		transferase
329	W58985	Homo sapiens adult brain clone BV141_2 encoded protein.	201	100
330	Y14455	Human secreted protein encoded by gene 45 clone HCFBJ91.	284	100
331	M25757	Bos taurus GTP:AMP phosphotransferase (EC 2.7.4.10)	127	73
332	AF037402	Bos taurus butyrophilin	225	25
333	X99211	Drosophila melanogaster ubiquitin-specific protease	846	68
334	Y36160	Human secreted protein #32.	500	100
335	W67869	Human secreted protein encoded by gene 63 clone HHGDB72.	454	91
336	X15334	Homo sapiens creatine kinase B	162	71
337	AL109658	Homo sapiens dJ776F14.1 (ortholog of mouse P47)	310	50
338	AL109658	Homo sapiens dJ776F14.1 (ortholog of mouse P47)	797	55
339	AJ001309	Homo sapiens DnaJ protein	659	100
340	AF096870	Homo sapiens estrogen-responsive B box protein	465	28
341	L33243	Homo sapiens polycystic kidney disease 1 protein	20117	99
342	Z83850	Homo sapiens mouse NIK serine threonine protein kinase like; match:	1582	95
		proteins P97820 CE02384
343	AF201084	Secale cereale secalin precursor	149	29
344	L16685	Caenorhabditis elegans homology with breakpoint cluster region protein;	549	41
		putative
345	AL162458	Homo sapiens bA465L10.4	2489	94
		(matrix metalloproteinase 9 (gelatinase B, 92 kD gelatinase,
		92 kD type IV collagenase) (CLG4B))
346	AX011001	Homo sapiens MMP-9	2375	100
347	X89416	Homo sapiens protein phosphatase 5	451	87
348	Y64786	Human 5′ EST related polypeptide SEQ ID NO: 947.	200	100
349	AC005167	Arabidopsis thaliana putative ubiquitin-conjugating enzyme	509	44
350	AF074901	Caenorhabditis elegans hemicentin precursor	1205	29
351	U12623	Rattus norvegicus cyclic nucleotide gated cation channel	2811	93
352	AF245516	Drosophila melanogaster Ran binding protein 9	140	40
353	AF300649	Homo sapiens regulator of G-protein signaling	926	100
354	X16396	Homo sapiens precursor polypeptide (AA −29 to 315)	855	80
355	S70290	Homo sapiens glutamine synthetase, GS {EC 6.3.1.2}	1787	100
356	X59834	Homo sapiens glutamate-ammonia ligase	435	89
357	X84801	Homo sapiens ZNF165	467	55
358	AL365234	Arabidopsis thaliana putative protein	224	34
359	AJ272034	Homo sapiens putative capacitative calcium channel	4470	100
360	AJ007798	Homo sapiens stromal antigen 3, (STAG3)	278	74
361	AJ007798	Homo sapiens stromal antigen 3, (STAG3)	292	82
362	AF193807	Homo sapiens Rh type B glycoprotein	2176	99
363	AC006963	Homo sapiens similar to Kelch proteins; similar to BAA77027	895	37
		(PID:g4650844)
364	Y86297	Human secreted protein HLDCE79, SEQ ID NO: 212.	530	100
365	AB039903	Homo sapiens interferon-responsive finger protein 1 long form	4302	99
366	AF143003	Perca flavescens lysyl oxidase related protein homolog	410	55
367	AF151840	Homo sapiens CGI-82 protein	1166	71
368	AB036834	Drosophila melanogaster MAP kinase phosphate	188	41
369	M30262	Homo sapiens preprocardiodilatin-atrial natriuretic factor	771	100
370	AF135253	Mus musculus fibulin-2	386	41
371	AF112361	Schmidtea mediterranea opsin	109	32
372	X89416	Homo sapiens protein phosphatase 5	387	100
373	M96256	Homo sapiens rapamycin binding protein	1169	100
374	AF121859	Homo sapiens sorting nexin 9	660	43
375	Y07566	Homo sapiens RIT (Ric-related gene expressed in many tissues)	266	28
376	AF208291	Homo sapiens protein kinase HIPK2	6242	99
377	AF170301	Mus musculus nuclear body associated kinase 1a	5972	97
378	AL133230	Homo sapiens dJ530I15.2	1170	99
		(novel protein similar to placental protein DIFF40)
379	AB001928	Homo sapiens cathepsin V	656	98
380	AB001928	Homo sapiens cathepsin V	725	90
381	Y87329	Human signal peptide containing protein HSPP-106 SEQ ID NO: 106.	692	94
382	AL356276	Homo sapiens bA367J7.2.1	479	51
		(novel Immunoglobulin domains containing protein (isoform 1))
383	AK024551	Homo sapiens unnamed protein product	824	100
384	U70851	Caenorhabditis elegans similar to S. cerevisiae protein transport protein	612	39
		SEC7 (SP:P11075)
385	D83348	Rattus norvegicus long type PB-cadherin	3904	92
386	AF162149	Mycoplasma bovis variable surface lipoprotein	216	46
387	U96149	Mus musculus perforatorial protein PERF 15	481	68
388	U12535	Homo sapiens epidermal growth factor receptor kinase substrate	1051	46
389	U00059	Saccharomyces cerevisiae Yhr116wp	114	41
390	U70369	Mus musculus hematopoietic-specific IL-2 deubiquitinating enzyme	873	47
391	AF247679	Xenopus laevis putative N-terminal acetyltransferase	978	57
392	AB007646	Arabidopsis thaliana UVB-resistance protein UVR8	315	31
393	AF026246	Homo sapiens HERV-E envelope glycoprotein	361	91
394	AF198489	Homo sapiens LBP-32	1747	57
395	Y91356	Human secreted protein sequence encoded by gene 11 SEQ ID NO: 77.	801	100
396	Y94978	Human secreted protein clone pw337_6 2nd protein sequence	444	100
		SEQ ID NO: 238.
397	AC005996	Homo sapiens similar to Xenopus laevis gamma-crystallin 6;	818	100
		similar to AF071563 (PID:g3930581)
398	Y76216	Human secreted protein encoded by gene 93.	225	97
399	Y11447	Human 5′ EST secreted protein SEQ ID No 269.	210	97
400	U12392	Haematobia irritans putative ATPase	917	49
401	U12392	Haematobia irritans putative ATPase	609	49
402	AF002109	Arabidopsis thaliana putative ABC transporter	487	40
403	AF110645	Homo sapiens candidate tumor suppressor p33 ING1 homolog	860	69
404	J03941	Mus musculus ferritin heavy chain	947	100
405	AL023513	Homo sapiens dJ268D13.1.1	808	46
		(seizure related gene 6 (mouse)-like (KIAA0927)
		(isoform 1))
407	D87458	Homo sapiens Similar to Human estrogen-responsive finger protein,	2575	99
		efp (A49656)
408	AJ278475	Homo sapiens transport-secretion protein 2.1 (TTS-2.1)	346	98
409	AF282886	Homo sapiens heparanase-like protein HPA2b	2785	100
410	AF015454	Xenopus laevis ER1	777	42
411	AF163762	Homo sapiens zinc metalloendopeptidase	5940	99
412	Z34277	Homo sapiens mucin	978	95
413	Y76179	Human secreted protein encoded by gene 56.	634	100
414	AF168362	Rattus norvegicus protein associating with small stress protein PASS1	232	68
415	AF156777	Homo sapiens ASB-1 protein	486	39
416	AK024169	Homo sapiens unnamed protein product	752	100
417	Y08625	Human secreted protein BL341 4.	367	100
418	AB026803	Mus musculus synaptotagmin VI	2153	96
419	AB026803	Mus musculus synaptotagmin VI	2098	94
420	AJ278474	Sus scrofa cytochrome P450	1316	50
421	W78135	Human secreted protein encoded by gene 10 clone HPNGQ80.	385	100
422	U70476	Rattus norvegicus cationic amino acid transporter-1	646	62
423	AF188634	Drosophila melanogaster F protein	546	55
424	AJ132889	Mus musculus kinesin like protein 9	3618	88
425	D86983	Homo sapiens similar to D. melanogaster peroxidasin (U11052)	328	32
426	AL035464	Homo sapiens dJ1043E3.1 (novel protein)	945	99
427	ALC34380	Homo sapiens dJ50O24.4	1131	67
		(novel protein with DHHC zinc finger domain)
428	AJ400877	Homo sapiens CEGP1 protein	5605	100
429	AF060570	Mus musculus rig-1 protein	1545	73
430	Y02697	Human secreted protein encoded by gene 48 clone HTNBR95.	165	100
431	Y73386	HTRM clone 3279329 protein sequence.	529	100
432	AF279890	Homo sapiens 2P domain potassium channel TREK2	2760	100
433	AB023658	Rattus norvegicus Ca/calmodulin-dependent protein	2468	93
		kinase kinase alpha, CaM-kiriase kinase alpha
434	J05056	Oryctolagus cuniculus oxysterol-binding protein	181	42
435	Y99437	Human PRO1508 (UNQ761) amino acid sequence SEQ ID NO: 336.	672	83
436	AF043179	Homo sapiens T cell receptor beta chain	681	73
437	AF151042	Homo sapiens HSPC208	585	79
438	Y87328	Human signal peptide containing protein HSPP-105 SEQ ID NO: 105.	681	100
439	Y66734	Membrane-bound protein PRO1097.	297	70
440	D78572	Mus musculus membrane glycoprotein	262	28
441	L20315	Mus musculus MPS1 protein	2716	77
442	Y19588	Amino acid sequence of a human secreted protein.	329	100
443	AF306550	Sinorhizobium meliloti (p)ppGpp synthetase	213	36
444	M38379	Arabidopsis thaliana calmodulin-1	108	37
445	Y12512	Human 5′ EST secreted protein SEQ ID NO: 543.	320	98
446	AC013482	Arabidopsis thaliana T26F17.15	309	37
447	AC013482	Arabidopsis thaliana T26F17.15	289	35
448	AL035703	Homo sapiens dJ61A9.1 (tyrosine kinase)	5326	99
449	X59720	Saccharomyces cerevisiae YCR017C, len: 953	731	30
450	AL035526	Arabidopsis thaliana extensin-like protein	154	33
451	U16282	Homo sapiens ELL	147	29
452	M80537	Drosophila melanogaster fat protein	207	28
453	AB029334	Halocynthia roretzi HrPET-1	687	38
454	AL031349	Schizosaccharomyces pombe putative vesicular transport protein	643	28
455	AB041533	Homo sapiens sperm antigen	3737	98
456	U27109	Homo sapiens prepromultimerin	208	24
457	AJ010949	Mus musculus calcium channel alpha-2-delta-C subunit	225	22
458	AJ271643	Homo sapiens putative acid-sensing ion channel	2893	100
459	G02479	Human secreted protein, SEQ ID NO: 6560.	442	100
460	D16111	Homo sapiens human homologue of rat phosphatidylethanolamine	846	98
		binding protein
461	AF180470	Mus musculus Kiaa0575	1139	57
462	Z54270	Caenorhabditis elegans F11C1.4	183	33
463	AB019527	Homo sapiens LDOC1 protein	129	31
464	U58105	Mus musculus Murine homolog of human ftp-3	303	36
465	AF172449	Homo sapiens opioid growth factor receptor	325	47
466	AL035106	Homo sapiens dJ998C11.1 (continues in Em:AL445192 as bA269H4.1)	2438	58
467	AC004561	Arabidopsis thaliana putative proline-rich protein	256	29
468	AF000198	Caenorhabditis elegans weak similarity to HSP90	144	28
469	X59720	Saccharomyces cerevisiae YCR016W, len: 290	128	26
470	AF027219	Homo sapiens ZNF202 beta	620	32
471	AF122924	Xenopus laevis Wnt inhibitory factor-1	484	44
472	X67704	Drosophila melanogaster sperm protein	111	32
473	AF317889	Homo sapiens NOX5	3015	99
474	L46815	Mus musculus DNA binding protein Rc	6027	77
475	AF190665	Mus musculus LMBR1 long form	2391	96
476	AF041382	Drosophila melanogaster microtubule binding protein D-CLIP-190	244	33
477	AL033545	Arabidopsis thaliana putative protein	138	49
478	Z98745	Homo sapiens dJ29K1.2	1123	56
479	AL118502	Homo sapiens bA371L19.1 (novel protein)	2471	100
480	Z94160	Homo sapiens dJ63G5.3 (putative Leucine rich protein)	953	100
481	X98263	Homo sapiens M-phase phosphoprotein 6	469	93
482	B08904	Human secreted protein sequence encoded by gene 14 SEQ ID NO: 61.	239	100
483	AY007233	Homo sapiens phosphoinositol 3-phosphate binding protein-1	3830	99
484	AY007233	Homo sapiens phosphoinositol 3-phosphate binding protein-1	4152	99
485	Y94947	Human secreted protein clone cw1292_8 protein sequence SEQ ID NO: 100.	408	100
486	J03614	Oryctolagus cuniculus myosin heavy chain	225	26
487	AB026256	Homo sapiens organic anion transporter OATP-B	188	92
488	AJ277748	Rattus norvegicus NTPDase6	501	96
489	U02289	Caenorhabditis elegans GTPase-activating protein	551	36
490	B08894	Human secreted protein sequence encoded by gene 4 SEQ ID NO: 51.	211	69
491	AF238862	Xenopus laevis Churchill protein	495	71
492	D87076	Homo sapiens similar to human bromodomain protein BR140 (JC2069)	107	30
493	A11959	synthetic construct PP4X	1122	99
494	Y87252	Human signal peptide containing protein HSPP-29 SEQ ID NO: 29.	391	100
495	AF015565	Dictyostelium discoideum VacA	245	31
496	AF305427	Mus musculus cAMP-dependent protein kinase regulatory subunit	940	78
497	K02581	Homo sapiens thymidine kinase (EC 2.7.1.21)	1216	99
498	AB048364	Mus musculus osteoblast differentiation promoting factor	1017	46
499	AJ006239	Homo sapiens dihydropteridine reductase	933	100
500	Y59795	Human normal ovarian tissue derived protein 72.	554	100
501	AL109965	Homo sapiens dJ1121G12.1.2	537	54
		(A novel protein containing a putative PHD finger domain, isoform 2)
502	Y12021	Human 5′ EST secreted protein SEQ ID NO: 334.	265	97
503	G01878	Human secreted protein, SEQ ID NO: 5959.	348	92
504	AL023777	Schizosaccharomyces pombe putative RNA-binding protein	318	39
505	M11902	Mus musculus proline-rich salivary protein	109	45
506	J01998	AKV murine leukemia virus gag-pol polyprotein	621	42
		(tag amber codon at 2250-2252 inserts Gln in Mo-MuLV)
507	AC012396	Arabidopsis thaliana tumor-related protein, putative	154	45
508	X06160	Homo sapiens precursor polypeptide (AA −24 to 140)	742	97
509	Y99351	Human PRO1481 (UNQ750) amino acid sequence SEQ ID NO: 41.	1725	100
510	M22760	Homo sapiens cytochrome c oxidase subunit Va	461	97
511	AF199008	Mus musculus PALS1	3396	96
512	AF145021	Mus musculus exportin 4	3814	98
513	AF119837	Cyprinus carpio hexokinase I	2020	75
514	Z69043	Homo sapiens translocon-associated protein delta subunit precursor	459	98
515	AF276512	Homo sapiens RNA polymerase II elongation factor ELL3	2071	99
516	M34379	Homo sapiens elastase/medullasin precursor (EC 3.4.21.37)	1320	96
517	Y11385	Human 5′ EST secreted protein SEQ ID No 207.	220	100
518	AF045022	Bos taurus phosphatidic acid-preferring phospholipase A1	4254	91
519	U00031	Caenorhabditis elegans Contains similarity to Pfam domain: PF00957	980	47
		(synaptobrevin), Score = 100.3, E-value = 1.2e-26, N = 1
520	X92098	Homo sapiens transmembrane protein	597	89
521	X92098	Homo sapiens transmembrane protein	1003	96
522	X57346	Homo sapiens HS1	1218	95
523	AF207901	Xenopus laevis cingulin	317	24
524	AL035678	Arabidopsis thaliana putative protein	246	36
525	Y30721	Amino acid sequence of a human secreted protein.	231	100
526	AL035659	Homo sapiens dJ979N1.1 (dJ979N1.1)	1822	47
527	AL035548	Schizosaccharomyces pombe putative ribose methyltransferase	263	38
528	AB015617	Homo sapiens ELKS	3464	72
529	L25665	Homo sapiens GTP-binding protein	2227	99
530	Y11794	Human 5′ EST secreted protein SEQ ID No: 394.	375	100
531	Y14685	Arabidopsis thaliana polynucleotide phosphorylase	1375	40
532	X52574	Mus musculus GTP binding protein	4571	91
533	AB040119	Homo sapiens mitochondrial import receptor Tom22	718	100
534	U23822	Danio rerio collagen II A1	295	43
535	U39621	Gallus gallus type V collagen	371	46
536	X79682	Felis catus neuronal protein	617	96
537	AF099935	Homo sapiens MDC-3.13 isoform 2	532	55
538	AF145672	Drosophila melanogaster BcDNA.GH12174	230	36
539	AL035709	Arabidopsis thaliana putative protein	572	37
540	AL022104	Schizosaccharcmyces pombe putative pre-mrna splicing factor rna helicase	431	31
541	AF113615	Homo sapiens FH1/FH2 domain-containing protein FHOS	1138	55
542	U85494	Zea mays LON1 protease	1797	47
543	X54162	Homo sapiens 64 Kd autoantigen	587	38
544	AB037158	Homo sapiens DSCR6a	165	40
545	Y07754	Human secreted protein fragment encoded from gene 11.	1723	100
546	AL009171	Drosophila melanogaster 62D9.o	184	33
547	M18963	Homo sapiens islet regenerating protein	817	89
548	U29463	Homo sapiens cytochrome b561	1125	91
549	Z25420	Gallus gallus class II INCENP protein	237	26
550	AL031324	Schizosaccharomyces pombe membrane atpase	1150	41
551	AF145609	Drosophila melanogaster BcDNA.GH02833	666	41
552	AF302079	Homo sapiens HSP22-like protein interacting protein 17	459	100
553	AJ242730	Homo sapiens polyhomeotic 2	451	59
554	U88167	Caenorhabditis elegans contains similarity to C2 domains	428	45
555	AF156779	Homo sapiens ASB-4 protein	513	33
556	AF305081	Homo sapiens tankyrase-related protein	5992	98
557	AB028968	Homo sapiens KIAA1045 protein	2090	99
558	AB028968	Homo sapiens KIAA1045 protein	1448	99
559	AJ250998	Mucor circinelloides carotenoid regulatory protein	381	40
560	AL109804	Homo sapiens dJ1009E24.1.1	6334	99
		(A novel protein similar to the mouse sialoadhesin, a macrophage sialic acid
		binding receptor, isoform 1)
561	Y76539	Human ovarian tumor EST fragment encoded protein 35.	261	98
562	L07765	Homo sapiens carboxylesterase	209	51
563	Z38061	Saccharomyces cerevisiae mal5, stal, len: 1367, CAI: 0.3,	206	21
		AMYH_YEAST P08640 GLUCOAMYLASE S1 (EC 3.2.1.3)
564	AY009133	Homo sapiens FYVE-finger-containing Rab5 effector protein Rabenosyn-5	4027	99
565	AF287478	Lytechinus variegatus embryonic blastocoelar extracellular matrix	1116	35
		protein precursor
566	AF223427	Xenopus laevis RRM-containing protein SEB-4	793	88
567	AP000693	Homo sapiens partial CDS	470	56
568	AJ131244	Homo sapiens Sec24A protein	5357	98
569	Y12049	Human 5′ EST secreted protein SEQ ID NO: 362.	253	92
570	AF255342	Homo sapiens putative pheromone receptor V1RL1 long form	1859	99
571	Y99355	Human PRO1295 (UNQ664) amino acid sequence SEQ ID NO: 54.	1265	100
572	ACC02397	Mus musculus C9	678	51
573	M28821	Mus musculus Tcte-1 peptide	1884	74
574	AF000196	Caenorhabditis elegans strong similarity to the SNF2/RAD54 family of	185	36
		helicases; partial CDS
575	L34587	Homo sapiens RNA polymerase II elongation factor SIII, p15 subunit	426	100
576	AL035419	Homo sapiens dJ1100H13.4 (putative RhoGAP domain containing protein)	212	35
577	D83004	Homo sapiens ubiquitin-conjugating enzyme E2 UbcH-ben	385	100
578	Y87271	Human signal peptide containing protein HSPP-48 SEQ ID NO: 48.	474	86
579	U96626	Mus musculus chondroadherin	341	29
580	Z85986	Homo sapiens dJ108K11.3	1468	99
		(similar to yeast suppressor protein SRP40)
581	AF061529	Mus musculus rjs	230	32
582	AF228738	Homo sapiens profilin IIa	750	100
583	AF228738	Homo sapiens profilin IIa	624	87
584	AF181640	Drosophila melanogaster BcDNA.GH09817	190	47
585	AB037901	Homo sapiens gene amplified in squamous cell carcinoma-1	1340	54
586	AF149285	Caenorhabditis elegans Osm-3	806	62
587	AF056021	Xenopus laevis p80 katanin	146	31
588	U80931	Caenorhabditis elegans strong similarity to class-III of pyridoxal-	866	49
		phoshate-dependent aminotransferases
589	AK026962	Homo sapiens unnamed protein product	1725	99
590	AB011483	Arabidopsis thaliana	288	28
591	AF152243	Mus musculus putative E1-E2 ATPase	3942	98
592	AJ243460	Leishmania major proteophosphoglycan	204	25
593	AX021519	Homo sapiens unnamed protein product	1317	99
594	AL121581	Homo sapiens dJ1022E24.4	1162	69
		(A novel protein weakly similar to protein-L-isoaspartate
		o-methyltransferase (EC 2.1.1.77))
595	AL031177	Homo sapiens dJ889M15.3 (novel protein)	177	44
596	AL035601	Arabidopsis thaliana putative protein	301	28
597	AF202118	Homo sapiens HOX D1 protein	1726	100
598	AC007228	Homo sapiens BC37295_1	955	42
599	Z69727	Schizosaccharomyces pombe probable ribosomal protein	249	34
600	L08134	Rattus norvegicus glycoprotein	278	24
601	AK025598	Homo sapiens unnamed protein product	3738	99
602	X89453	Rattus norvegicus DRPLA	173	27
603	AF137030	Homo sapiens transmembrane protein 2	1285	57
604	AL356014	Arabidopsis thaliana putative protein	436	44
605	G03490	Human secreted protein, SEQ ID NO: 7571.	450	96
606	AY007380	Homo sapiens F-box protein FBX30	1211	100
607	D38076	Homo sapiens Ran-BP1 (Ran-binding protein 1)	916	100
608	U27837	Diphyllobothrium dendriticum actin	1033	50
609	M25750	Oryctolagus cuniculus sarcolumenin precursor	2307	97
610	AF180920	Homo sapiens cyclin L ania-6a	630	51
611	AF168990	Homo sapiens putative GTP-binding protein	2742	99
612	U77942	Homo sapiens syntaxin 7	421	98
613	U00046	Caenorhabditis elegans similar to yeast heat shock protein STI1	362	27
614	AB001928	Homo sapiens cathepsin V	226	100
615	AB016687	Arabidopsis thaliana SMC-like protein	326	22
616	AL163279	Homo sapiens homolog to cAMP response element binding and	181	53
		beta transducin family proteins
617	AF176524	Mus musculus F-box protein FBL10	525	50
618	AF070656	Homo sapiens HSPC002	731	86
619	Y00339	Homo sapiens carbonic anhydrase II (AA 1-260)	910	88
620	M29913	Homo sapiens eosinophil peroxidase	3757	100
621	AC006768	Caenorhabditis elegans contains similarity to Mycoplasma genitalium	1216	54
		glycerol-3-phospate dehydrogenase (SW: P47285)
622	AB033595	Mus musculus gasdermin	369	31
623	AF132449	Mus musculus smoothelin small isoform S1	646	54
624	AF247039	porcine adenovirus 3 163R*	141	31
625	L43065	Saccharomyces cerevisiae suppresses the respiratory deficiency of a yeast	403	36
		pet mutant
626	AF071172	Homo sapiens HERC2	483	35
627	Y40090	Peptide sequence derived from a human secreted protein.	921	98
628	AP001306	Arabidopsis thaliana contains similarity to cell wall-plasma membrane	717	36
		linker protein-gene_id: MKA23.5
629	AE004543	Pseudomonas aeruginosa probable MFS transporter	207	32
630	AK025204	Homo sapiens unnamed protein product	1925	99
631	AL050321	Homo sapiens dJ717M23.1 (novel gene)	2443	98
632	AL050321	Homo sapiens dJ717M23.1 (novel gene)	3988	99
634	U87804	Caulobacter crescentus GidA	396	65
635	AF004161	Oryctolagus cuniculus peroxisomal Ca-dependent solute carrier	154	52
636	U78090	Rattus norvegicus potassium channel regulator 1	682	91
637	AC002394	Homo sapiens Gene product with similarity to dynein beta subunit	536	41
638	AL109640	Homo sapiens dJ543J19.5 (CGI-107 Protein)	478	98
639	AB017065	Arabidopsis thaliana contains similarity to small nuclear	125	34
		ribonucleoprotein-gene id: MFC16.18
640	AK024498	Homo sapiens FLJ00106 protein	927	99
641	AF187064	Homo sapiens p75NTR-associated cell death executor; NADE	303	57
642	AB017507	Homo sapiens Apg12	724	100
643	AF035632	Rattus norvegicus syntaxin 12	1231	90
644	U48852	Cricetulus griseus HT protein	974	57
645	AB015798	Homo sapiens DnaJ homolog	1572	93
646	AB031046	Homo sapiens HMG-box transcription factor TCF-3	3096	100
647	U78547	Chlamydomonas reinhardtii PF20	209	38
648	AK024452	Homo sapiens FLJ00044 protein	2019	63
649	Y13117	Human secreted protein encoded by 5′ EST SEQ ID NO: 131.	268	100
650	AB023419	Mus musculus mSox7	1803	87
651	AF176688	Rattus norvegicus sodium/calcium/potassium exchanger NCKX1	184	34
652	AK026622	Homo sapiens unnamed protein product	4385	99
653	AFC16712	Mus musculus testicular condensing enzyme	191	36
654	AF135440	Mus musculus huntington yeast partner C	4306	94
655	AF135440	Mus musculus huntington yeast partner C	3709	86
656	AK022184	Homo sapiens unnamed protein product	680	100
657	U02467	Lilium longiflorum meiotin-1	182	33
658	M17099	Oryctolagus cuniculus progesterone-induced protein	1832	93
659	AF113132	Homo sapiens phosphoserine aminotransferase	1673	100
660	AF294790	Mus musculus RING-finger protein MURF	137	27
661	X56203	Plasmodium falciparum liver stage antigen	157	22
662	Y64994	Human 5′ EST related polypeptide SEQ ID NO: 1155.	372	100
663	U82626	Rattus norvegicus basement membrane-associated chondroitin proteoglycan	146	22
		Bamacan
664	AF155739	Mus musculus axotrophin	3007	85
665	AB037596	Mus musculus beta-1,6-N-acetylglucosaminyltransferase B	1257	73
666	AF168132	Homo sapiens RU1	2485	55
667	AF132961	Homo sapiens CGI-27 protein	1197	100
668	AB025258	Mus musculus granuphilin-a	729	42
669	AL391688	Homo sapiens bA524D16A.2.1	762	43
		(novel protein similar to mouse granuphilin-a)
670	AP001745	Homo sapiens human cDNA DKFZp586F0422, Accession No. AL050173	3747	100
671	X58454	Homo sapiens DS dopamine receptor	1024	85
672	AB017059	Arabidopsis thaliana FH protein interacting protein FIP2	212	35
673	AF255908	Streptococcus pneumoniae PspA	177	29
674	Y02669	Human secreted protein encoded by gene 20 clone HMKAH10.	288	100
675	AB024986	Oryza sativa cyclin	142	48
676	X70681	Xenopus laevis zinc finger protein	144	33
677	AL390026	Homo sapiens dJ336K20B.1 (novel protein based on FGENESH)	964	100
678	AC026815	Oryza sativa putative ATP-dependent RNA helicase (5′-partial)	437	43
679	AF196779	Homo sapiens JM10 protein	1182	98
680	U40802	Caenorhabditis elegans similar to other protein phosphatases 1, 2A and 2B	325	36
681	AJ238854	Rattus norvegicus type A/B hnRNP p40	1659	91
682	Y36083	Extended human secreted protein sequence, SEQ. ID NO. 468.	430	98
683	U37283	Homo sapiens microfibril-associated glycoprotein-2 MAGP-2	846	94
684	AJ235270	Rickettsia prowazekii 50S RIBOSOMAL PROTEIN L10 (rplJ)	122	23
685	W67828	Human secreted protein encoded by gene 22 clone HFEAF41.	510	100
686	W67828	Human secreted protein encoded by gene 22 clone HFEAF41.	403	85
687	AK026226	Homo sapiens unnamed protein product	1937	100
688	AK026226	Homo sapiens unnamed protein product	578	100
689	D26549	Bos taurus bovine adseverin	3527	92
690	AK024644	Homo sapiens unnamed protein product	245	67
691	X63546	Homo sapiens oncogene	1083	78
692	X63546	Homo sapiens oncogene	1613	83
693	AB029334	Halocynthia roretzi HrPET-1	614	37
694	AF286473	Mus musculus retinitis pigmentosa GTPase regulator	181	35
695	AB002405	Homo sapiens LAK-4p	346	34
696	Y14448	Human secreted protein encoded by gene 38 clone HFGAH44.	316	100
697	AL133216	Homo sapiens bA291L22.2	256	100
		(similar to CDC10 (cell division cycle 10, S. cerevisiae , homolog))
698	AF065389	Homo sapiens tetraspan NET-4	1111	74
699	AF161390	Homo sapiens HSPC272	281	81
700	Z83114	Caenorhabditis elegans K09B11.2	233	37
701	Y41360	Human secreted protein encoded by gene 53 clone H3PAD75.	490	100
702	Y41360	Human secreted protein encoded by gene 53 clone HJPAD75.	233	92
703	AL049481	Arabidopsis thaliana putative protein	829	53
704	U67949	Caenorhabditis elegans contains similarity t to sugar and other	486	36
		transporters (Pfam: sugar_tr.hmm.score: 13.46)
705	J02883	Homo sapiens colipase precursor	243	100
706	X66286	Gallus gallus tensin	1009	73
707	G01118	Human secreted protein, SEQ ID NO: 5199.	360	100
708	D50857	Homo sapiens DOCK180 protein	1060	51
709	AE003511	Drosophila melanogaster CG14194 gene product	542	54
710	L26247	Homo sapiens isolog of yeast sui1 and rice gos2; putative	470	86
711	Y12952	Amino acid sequence of a human secreted peptide.	362	98
712	U41849	Saccharomyces cerevisiae Yta6p	156	23
713	AF168362	Rattus norvegicus protein associating with small stress protein PASS1	220	35
714	Y48256	Human prostate cancer-associated protein 42.	242	75
715	AF053130	Mus musculus unconventional myosin MYO15	579	36
716	AF083385	Homo sapiens 30 kDa splicing factor; SPF 30	133	40
717	AF182417	Homo sapiens MDS016	406	100
718	AF065389	Homo sapiens tetraspan NET-4	584	43
719	W78132	Human secreted protein encoded by gene 7 clone HPEBD85.	246	100
720	AK023971	Homo sapiens unnamed protein product	1358	100
721	AF288813	Mus musculus synembryn	2384	86
722	X56203	Plasmodium falciparum liver stage antigen	188	19
723	AF190501	Homo sapiens leucine-rich repeat-containing G protein-coupled receptor 6	4286	99
724	AF022789	Homo sapiens ubiquitin hydrolyzing enzyme I	1689	88
725	AF176531	Mus musculus F-box protein FBX16	1412	80
726	U10991	Homo sapiens G2	756	30
727	AE001795	Thermotoga maritima glycerol kinase	845	38
728	AJ006692	Homo sapiens ultra high sulfer keratin	868	71
729	Y76177	Human secreted protein encoded by gene 54.	866	99
730	AL162295	Arabidopsis thaliana guanine nucleotide exchange factor-like protein	261	24
731	AF016036	Drosophila tsacasi repressor-like protein	220	27
732	Z54327	Caenorhabditis elegans contains similarity to Pfam domain: PF00070	339	32
		(pyridine nucleotide-disulphide oxidoreductase),
		Score = 82.7, E-value = 4e-24, N = 2
733	AC011713	Arabidopsis thaliana Contains similarity to gb|AF092102 G-protein	303	26
		beta subunit git5p from Schizosaccharomyces pombe and contains
		2 PF|00400WD doinain, G-beta repeat domains.
734	AL109965	Homo sapiens dJ1121G12.1.2	323	60
		(A novel protein containing a putative PPD finger domain, isoform 2)
735	AB000910	Sus scrofa ribosomal protein	483	87
736	Y07931	Human secreted protein fragment encoded from gene 80.	213	100
737	M95718	Oryctolagus cuniculus keratin	233	65
738	AL034382	Schizosaccharomyces pombe putative Trp-Asp repeat protein	453	35
739	Y38401	Human secreted protein encoded by gene No. 16.	233	88
740	U18917	Saccharomyces cerevisiae Yer157wp	338	26
741	W64471	Human secreted protein from clone DF989_3.	545	97
742	AJ000474	Homo sapiens cytidine deaminase	389	100
743	AL132954	Arabidopsis thaliana putative protein	361	34
744	AF026292	Homo sapiens chaperonin containing t-complex polypeptide 1,	890	100
		eta subunit; CCT-eta
745	Y60251	Human endometrium tumour EST encoded protein 311.	392	98
746	U07817	Dictyostelium discoideum glutamine-asparagine rich protein	148	30
747	M58529	Homo sapiens pro-alpha-2 type V collagen	158	100
748	AL118506	Homo sapiens dJ591C20.5 (KIAA1196)	952	45
749	AF181645	Drosophila melanogaster BcDNA.GH12144	105	54
750	AF067136	Homo sapiens protein phosphatase-1 regulatory subunit 7 alpha2	147	38
751	AJ006973	Homo sapiens TOM1	638	82
752	AF145664	Drosophila melanogaster BcDNA.GH11110	766	37
753	Z70310	Caenorhabditis elegans contains similarity to Pfam domain: PF00013	510	34
		(KH domain), Score = 42.8, E-value = 3.7e-12, N = 1;
		PF00023 (Ank repeat), Score = 428.2, E-value = 2.4e-125, N = 19
754	AF276514	Mus musculus 105-kDa kinase-like protein	3769	88
755	Y30734	Amino acid sequence of a human secreted protein.	258	96
756	AF102129	Rattus norvegicus KPL2	1436	79
757	L15313	Caenorhabditis elegans putative	829	58
758	AE003584	Drosophila melanogaster CG7289 gene product	529	34
759	Y66693	Membrane-bound protein PRO1004.	569	92
760	AC007766	Homo sapiens R26610_1	1574	80
761	AF073958	Homo sapiens cytokine-inducible SH2 protein 6	1142	54
762	Z71181	Caenorhabditis elegans contains similarity to Pfam domain: PF00561	498	34
		(alpha/beta hydrolase fold)
		Score = 83.7, E-value = 1.2e-21, N = 1
763	AC005724	Arabidopsis thaliana putative C3HC4-type RING zinc finger protein	166	45
764	AF052433	Strongylocentrotus purpuratus katanin p80 subunit	170	30
765	X56044	Mus musculus protein Htf9C	759	44
766	U56418	Homo sapiens lysophosphatidic acid acyltransferase-beta	834	100
767	AK024388	Homo sapiens unnamed protein product	675	100
768	W67828	Human secreted protein encoded by gene 22 clone HFEAF41.	278	100
769	AF055993	Homo sapiens mSin3A associated polypeptide p30	637	71
770	AK024512	Homo sapiens unnamed protein product	454	93
771	AF095844	Homo sapiens melanoma differentiation associated protein-5	687	40
772	AF169257	Homo sapiens sodium/calcium exchanger NCKX3	3084	99
773	D00824	Gallus gallus alpha 1 chain of type XII collagen	302	30
774	X82557	Rattus norvegicus myelin and lymphocyte protein	257	39
775	G03200	Human secreted protein, SEQ ID NO: 7281.	548	98
776	AK024793	Homo sapiens unnamed protein product	1730	100
777	AB019397	Homo sapiens DNA topoisomerase II binding protein	108	23
778	AK025033	Homo sapiens unnamed protein product	817	99
779	AB028069	Homo sapiens activator of S phase Kinase	474	36
780	AF273052	Homo sapiens CTCL tumor antigen se70-2	1294	100
781	AF217512	Homo sapiens uncharacterized bone marrow protein BM036	130	100
782	AF121857	Homo sapiens sorting nexin 7	2019	100
783	AF305941	Homo sapiens LIM2	933	100
784	AF153085	Homo sapiens phosphoprotein pp75	178	31
785	AF106584	Caenorhabditis elegans contains similarity to homeobox domains	255	32
		(Pfam: PF00046, score = 16.2, E = 0.015, N = 1)
786	AF106584	Caenorhabditis elegans contains similarity to homeobox domains	244	31
		(Pfam: PF00046, score = 16.2, E = 0.015, N = 1)
787	AJ278508	Mus musculus MDM2 binding protein	567	76
788	Z21507	Homo sapiens human elongation factor-1-delta	269	96
789	AF096286	Mus musculus pecanex 1	1498	52
790	AC004882	Homo sapiens similar to calmodulin; similar to P24044 (PID: g115520)	525	60
791	AJ133500	Xenopus laevis p33 ringo	547	49
792	U41264	Caenorhabditis elegans coded for by C. elegans CDNA cm13gl;	212	50
		Similar to bumetanide-sensitive Na—K—Cl cotransporter
793	X16078	Torpedo californica 4-acetamido-4′-isothiocyanostilbene-2,2′-disulphonic	423	38
		acid-binding protein
794	AK026447	Homo sapiens unnamed protein product	833	100
795	AL035417	Homo sapiens dJ891H21.1 (HYPOTHETICAL 43.1 KD PROTEIN)	465	100
796	M83751	Homo sapiens arginine-rich protein	525	62
797	AB037834	Homo sapiens KIAA1413 protein	7273	99
798	AB037834	Homo sapiens KIAA1413 protein	6449	98
799	AC011717	Arabidopsis thaliana putative carnitine/acylcarnitine translocase;	450	37
		50581-51656
800	AC069143	Arabidopsis thaliana Contains similarity to a transposable element Tip100	305	25
		protein for transposase from Ipomoea purpurea gb|4063769 and is a
		member of the transmembrane 4 family PF|00335.
801	AY008372	Homo sapiens oxysterol binding protein-related protein 3	4671	100
802	AC004557	Arabidopsis thaliana F17L21.20	404	39
803	AF182218	Homo sapiens epidermal lipoxygenase	3830	99
804	U90353	Strongyloides stercoralis IgG and IgE immuncreactive antigen recognized	108	35
		by sera from patients with strongyloidiasis
805	D13989	Homo sapiens human rho GDI	818	99
806	U00482	Homo sapiens gamma-subunit of rod cGMP-phosphodiesterase	472	100
807	Y19599	SEQ ID NO 317 from WO9922243.	590	98
808	D50617	Saccharomyces cerevisiae YFL042C	227	31
809	X58430	Homo sapiens homeobox protein	478	100
810	M98539	Homo sapiens prostaglandin D2 synthase	464	97
811	AJ004872	Homo sapiens TCR beta chain	1296	92
812	G00329	Human secreted protein, SEQ ID NO: 4410.	524	100
813	AL031431	Homo sapiens dJ462O23.2 (novel protein)	824	45
814	M37760	Mus musculus serine 2 ultra high sulfur protein	633	55
815	AL121673	Homo sapiens bA305P22.2 (novel protein)	2396	100
816	AB027568	Mus musculus thiamin pyrophosphokinase	664	91
817	M37190	Homo sapiens ras inhibitor	792	40
818	AL109659	Homo sapiens dJ1024N4.1	2858	100
		(novel Sodium:solute symporter family member similar to SLC5A1
		(SGLT1))
819	AB000170	Sus scrofa endopeptidase 24.16 type M1	3472	94
820	AF058789	Rattus norvegicus SynGAP-a	6651	99
821	AK024408	Homo sapiens unnamed protein product	1040	100
822	Y08260	Mus musculus cytoplasmic polyadenylation element-binding protein (CPEB)	2453	95
823	Y36090	Extended human secreted protein sequence, SEQ ID NO. 475.	420	95
824	AC003682	Homo sapiens R28530_2	1398	48
825	D89052	Homo sapiens proton-ATPase-like protein	356	100
826	AB039903	Homo sapiens interferon-responsive finger protein 1 long form	978	61
827	U96963	Mus musculus p140mDia	471	26
828	AJ251641	Mus musculus syncoilin	656	89
829	Z98595	Schizosaccharomyces pombe coronin-like protein	196	24
830	AF116660	Homo sapiens PRO1430	299	100
831	AJ277442	Homo sapiens xylosyltransferase II	4625	100
832	D86081	Mus musculus S-II-T1	290	36
833	L21671	Mus musculus Eps8	547	31
834	Z34801	Caenorhabditis elegans Similarity with drosphila MSP-300 protein	283	25
		(PIR acc. no. S30431) , contains similarity to Pfam domain: PF01465
		(GRIP domain), Score = 90.2, E-value = 1.4e-23, N = 1
835	AF264750	Homo sapiens ALR-like protein	21361	100
836	AF176814	Mus musculus Ab1-philin 2	247	41
837	AL121771	Homo sapiens dJ548G19.1.2	166	24
		(novel protein (ortholog of mouse zinc finger protein ZFP64)
		(translation of cDNA NT2RP4001938 (Em: AK001744)) (isoform 2))
838	G00405	Human secreted protein, SEQ ID NO: 4486.	397	98
839	AF130357	Mus musculus domesticus similar to RNA binding protein	222	53
840	AJ272034	Homo sapiens putative capacitative calcium channel	4470	100
841	G02337	Human secreted protein, SEQ ID NO: 6418.	377	98
842	AF286475	Takifugu rubripes retinitis pigmentosa GTPase regulator-like protein	226	22
843	AB030183	Mus musculus contains transmembrane (TM) region	1159	89
844	Y18890	Human endogenous retrovirus K gag protein	273	47
845	AC004890	Homo sapiens similar to HUB1; similar to BAA24380 (PID: g2789430)	435	79
846	Y66666	Membrane-bound protein PRO1013.	1858	99
847	Y66666	Membrane-bound protein PRO1013.	1658	93
848	AF119913	Homo sapiens PRO3077	620	100
849	Y36233	Human secreted protein encoded by gene 10.	302	96
850	D14530	Homo sapiens ribosomal protein	341	100
851	AC011713	Arabidopsis thaliana Is a member of the PP|01553 Acyltransferase family.	162	39
852	Y36154	Human secreted protein #26.	296	98
853	AC004877	Homo sapiens zinc finger-like; similar to P52742 (PID: g1731411)	2830	100
854	M34163	Mus musculus low affinity IgE receptor (FC-epsilon-RII)	293	29
855	X73579	Rattus norvegicus CD23	333	31
856	M34427	Homo sapiens T-plastin	414	98
857	Y19456	Amino acid sequence of a human secreted protein.	307	90
858	AF067136	Homo sapiens protein phosphatase-1 regulatory subunit 7 betal	199	47
859	AC023279	Arabidopsis thaliana F12K21.21	441	38
860	G04069	Human secreted protein, SEQ ID NO: 8150.	491	100
861	AP001297	Arabidopsis thaliana gene _id:F14O13.28˜	488	28
862	Y19587	Amino acid sequence of a human secreted protein.	160	96
863	AB039861	Schizosaccharomyces pombe Esol	201	31
864	U34932	Rattus norvegicus Fos-related antigen	2062	84
865	Y60152	Human endometrium tumour EST encoded protein 212.	379	100
866	AK027028	Homo sapiens unnamed protein product	2294	99
867	AF039718	Caenorhabditis elegans contains similarity to lupus LA protein homologs	329	44
868	AK024480	Homo sapiens FLJ00074 protein	1001	100
869	AF320909	Homo sapiens MAGE-E1	634	100
870	AF126484	Homo sapiens CARD4	181	30
871	Z83844	Homo sapiens dJ37E16.5	651	46
		(novel protein similar to nitrophenylphosphatases from various organisms)
872	AF128406	Homo sapiens nuclear prelamin A recognition factor	1121	47
873	Y16790	Homo sapiens keratin type I	2029	99
874	AJ006692	Homo sapiens ultra high sulfer keratin	1073	88
875	U43281	Saccharomyces cerevisiae Lpg21p	323	40
876	AF078844	Homo sapiens hqp0376 protein	362	81
877	AB025258	Mus musculus granuphilin-a	3193	90
878	D37991	Homo sapiens beta-signal sequence receptor	847	99
879	AL035427	Homo sapiens dJ769N13.1 (KIAA0443 protein.)	510	28
880	AL109925	Homo sapiens dJ534K7.2 (novel protein)	3323	100
881	AF254411	Homo sapiens ser/arg-rich pre-mRNA splicing factor SR-A1	6833	100
882	AL137082	Arabidopsis thaliana putative protein	310	42
883	M97188	Strongylocentrotus purpuratus tektin A1	292	40
884	Y10837	Amino acid sequence of a human secreted protein.	434	100
885	Z12172	Homo sapiens putative homeotic protein	1388	79
886	AJ000479	Homo sapiens putative G-Protein coupled receptor, EDG6	177	30
887	X56044	Mus musculus protein Htf9C	2360	75
888	AC006963	Homo sapiens similar to Kelch proteins; similar to BAA77027 (PID: g4650844)	268	27
889	AF205357	Drosophila melanogaster extracellular matrix protein papilin	503	33
890	AB027757	Cicer arietinum NADPH oxidoreductase homolog	719	45
891	AL022140	Arabidopsis thaliana serine/threonine protein kinase like protein	153	30
892	AF286473	Mus musculus retinitis pigmentosa GTPase regulator	208	28
893	AL050328	Homo sapiens bA145L22.2	2072	99
		(novel KRAB box containing C2H2 type zinc finger protein)
894	AL096856	Arabidopsis thaliana putative protein	223	44
895	AF245517	Homo sapiens vacuolar proton pump 116 kDa accessory subunit	4378	100
896	AB000113	Rattus norvegicus cationic amino acid transporter 3	2644	83
897	AF142406	Babesia bigemina 200 kDa antigen p200	588	24
898	AF062476	Mus musculus retinoic acid-responsive protein; STRA6	2184	75
899	Y48507	Human breast tumour-associated protein 52.	308	98
900	U79745	Homo sapiens monocarboxylate transporter homologue MCT6	465	46
901	AF145661	Drosophila melanogaster BcDNA.GH10646	376	24
902	AC009519	Arabidopsis thaliana F1N19.17	241	29
903	Y27576	Human secreted protein encoded by gene No. 10.	394	96
904	AF192968	Homo sapiens high-glucose-regulated protein 8	2041	67
905	AP001743	Homo sapiens putative gene, ankirin like, possible dual specifity	4306	99
		Ser/Thr/Tyr kinase domain
906	U71205	Mus musculus rit	252	35
907	M77003	Mus musculus glycerol-3-phosphate acyltransferase	3366	93
908	AF286368	Homo sapiens eppin-1	222	54
909	AC006439	Arabidopsis thaliana putative ADP-ribosylation factor	261	34
910	AF230808	Homo sapiens zinc finger transcription factor Pegasus	2279	99
911	AF134804	Mus musculus putative zinc finger transcription factor OVO1	698	52
912	AF049907	Homo sapiens zinc finger transcription factor	530	31
913	AF217319	Mus musculus putative repair and recominbination helicase RAD26L	3218	89
914	X80473	Mus musculus rab19	1008	88
915	U88315	Caenorhabditis elegans weak similarity to Plasmodium vivax	609	36
		reticulocyte-binding protein 2 (GI: 160628)
916	AF273052	Homo sapiens CTCL tumor antigen se70-2	1237	99
917	U34932	Rattus norvegicus Fos-related antigen	2258	84
918	AF116911	Mus musculus thymic dendritic cell-derived factor 1	305	92
919	AF205935	Mus musculus MGA protein	4822	84
920	AL157413	Homo sapiens bA526K17.1 (novel protein).	1076	100
921	Y10823	Amino acid sequence of a human secreted protein.	183	100
922	Z97653	Homo sapiens c380A1.1b (novel protein)	1106	100
923	Z24725	Homo sapiens mitogen inducible gene mig-2	1461	56
924	AF113917	Homo sapiens NADP+-dependent isocitrate dehydrogenase	1428	100
925	AF064604	Homo sapiens KE03 protein	1025	57
926	AF064604	Homo sapiens KE03 protein	972	58
927	AL033545	Arabidopsis thaliana putative protein	84	35
928	AJ007014	Homo sapiens AMMECR1 protein	733	64
929	AJ276316	Homo sapiens zinc finger protein 304	1007	51
930	AC004077	Arabidopsis thaliana putative katanin	877	57
931	D16226	Oryctolagus cuniculus one of the members of sodium-glucose	2008	71
		cotransporter family
932	D16226	Oryctolagus cuniculus one of the members of sodium-glucose	3026	83
		cotransporter family
933	AF234676	Sus scrofa adipose differentiation-related protein	336	29
934	U70732	Homo sapiens glutamate pyruvate transaminase	1821	68
935	AB027004	Homo sapiens protein phosphatase	462	48
936	AF043180	Homo sapiens T cell receptor beta chain	1097	69
937	AC006528	Arabidopsis thaliana putative DNA replication licensing factor	742	41
938	L08483	Drosophila melanogaster ring canal protein	660	31
939	AL022238	Homo sapiens dJ1042K10.4 (novel protein)	758	46
940	AF275151	Rattus norvegicus androgen receptor-related apoptosis-associated protein	619	55
		CBL27
941	AF275151	Rattus norvegicus androgen receptor-related apoptosis-associated protein	619	55
		CBL27
942	AE000854	Methanothermobacter thermoautotrophicus Na+/H+-exchanging protein:	498	35
		Na+/H+ antiporter
943	D88894	Homo sapiens brain acyl-CoA hydrolase	1742	100
944	AP001072	Oryza sativa Similar to Arabidopsis thaliana DNA chromosome 4,	248	33
		BAC clone F22K18, putative protein. (AL035356)
945	AB033744	Mus musculus type II cytokeratin	835	74
946	AP186461	Rattus norvegicus ring finger protein Fxy	351	30
947	AK025539	Homo sapiens unnamed protein product	2555	99
948	J03407	Homo sapiens rfp transforming protein	507	40

TABLE 3
SEQ ID NO:	ACCESSION NO.	DESCRIPTION	RESULTS*
2	BL00972	Ubiquitin carboxyl-	BL00972A 11.93 2.500e-20 267-285
		terminal hydrolases	BL00972D 22.55 5.179e-17 828-853
		family 2 proteins.	BL00972E 20.72 8.650e-13 855-877
			BL00972C 16.48 7.120e-11 411-426
			BL00972B 9.45 7.923e-10 353-363
3	BL00018	EF-hand calcium-	BL00018 7.41 5.696e-09 331-344
		binding domain proteins.
4	BL00018	EF-hand calcium-	BL00018 7.41 5.696e-09 331-344
		binding domain proteins.
5	PR00401	SH2 DOMAIN SIGNATURE	PR00401B 12.94 1.000e-08 340-351
6	PR00401	SH2 DOMAIN SIGNATURE	PR00401B 12.94 1.000e-08 367-378
7	BL00625	Regulator of chromosome	BL00625A 16.21 7.787e-16 308-337
		condensation (RCC1)	BL00625A 16.21 7.369e-15 190-219
		proteins.	BL00625B 17.69 1.514e-13 302-336
			BL00625B 17.69 2.286e-13 184-218
			BL00625B 17.69 3.957e-13 132-166
			BL00625A 16.21 5.690e-13 138-167
			BL00625A 16.21 5.731e-11 360-389
			BL00625B 17.69 3.333e-10 354-388
9	BL00120	Lipases, serine proteins.	BL00120B 11.37 9.486e-12 166-181
15	BL00183	Ubiquitin-conjugating	BL00183 28.97 1.329e-10 45-93
		enzymes proteins.
18	PR00049	WILM'S TUMOUR	PR00049D 0.00 6.034e-09 262-277
		PROTEIN SIGNATURE
19	PR00049	WILM'S TUMOUR	PR00049D 0.00 6.034e-09 74-89
		PROTEIN SIGNATURE
20	BL00790	Receptor tyrosine	BL00790D 12.41 8.297e-09 804-829
		kinase class V proteins.
21	BL00790	Receptor tyrosine	BL00790D 12.41 8.297e-09 878-903
		kinase class V proteins.
23	PR00380	KINESIN HEAVY CHAIN	PR00380D 9.93 2.080e-22 321-343
		SIGNATURE	PR00380A 14.18 1.486e-21 79-101
			PR00380B 12.64 6.571e-18 217-235
			PR00380C 13.18 6.927e-13 269-288
25	BL01242	Formamidopyrimidine-	BL01242F 17.92 5.300e-11 32-66
		DNA glycosylase proteins.
27	PF00651	BTB (also known as BR-C/Ttk)	PF00651 15.00 2.500e-14 46-59
		domain proteins.
28	DM00215	PROLINE-RICH PROTEIN 3.	DM00215 19.43 3.898e-09 99-132
29	PR00380	KINESIN HEAVY CHAIN	PR00380A 14.18 9.250e-25 93-115
		SIGNATURE	PR00380D 9.93 4.857e-19 302-324
			PR00380B 12.64 4.429e-18 212-230
			PR00380C 13.18 1.692e-16 247-266
30	BL00107	Protein kinases ATP-binding	BL00107A 18.39 3.368e-18 36-67
		region proteins.
32	BL00594	Aromatic amino acids	BL00594A 16.75 9.376e-09 76-120
		permeases proteins.
34	BL00790	Receptor tyrosine	BL00790E 29.58 1.111e-12 614-662
		kinase class V proteins.	BL00790E 29.58 3.111e-12 668-716
			BL00790E 29.58 7.000e-10 560-608
38	BL00290	Immunoglobulins and	BL00290A 20.89 4.150e-12 126-149
		major histocompatibility
		complex proteins.
39	PR00019	LEUCINE-RICH REPEAT	PR00019A 11.19 6.087e-10 93-107
		SIGNATURE	PR00019B 11.36 7.840e-09 90-104
42	PD01443	INHIBITOR CALPAIN	PD01443D 8.36 4.670e-09 815-837
		CALPASTATIN REPEAT
		THIOL PROT.
43	BL01101	Casein kinase II	BL01101A 16.07 1.000e-40 9-54
		regulatory subunit proteins.	BL01101B 10.94 9.000e-31 72-97
46	BL01166	RNA polymerases beta	BL01166G 18.10 2.500e-34 824-866
		chain proteins.	BL01166H 19.05 9.410e-30 936-986
			BL01166D 17.37 4.396e-19 612-642
			BL01166E 13.47 8.244e-17 682-706
			BL01166C 12.21 9.357e-12 431-456
47	BL00518	Zinc finger, C3HC4 type	BL00518 12.23 7.000e-09 25-34
		(RING finger), proteins.
48	BL00038	Myc-type, ′helix-	BL00038A 13.61 6.625e-11 284-300
		loop-helix′
		dimerization domain proteins.
49	BL00905	GTP1/OBG family proteins.	BL00905D 15.00 4.214e-10 125-140
50	BL00107	Protein kinases ATP-	BL00107B 13.31 7.300e-15 64-80
		binding region proteins.
53	BL00383	Tyrosine specific	BL00383E 10.35 5.263e-09 328-339
		protein phosphatases proteins.
55	BL00383	Tyrosine specific	BL00383E 10.35 5.263e-09 246-257
		protein phosphatases proteins.
56	BL00383	Tyrosine specific	BL00383E 10.35 5.263e-09 328-339
		protein phosphatases proteins.
62	BL00226	Intermediate filaments proteins.	BL00226B 23.86 5.919e-09 560-608
64	PR00322	G10 PROTETN	PR00322E 6.62 1.720e-10 30-40
65	BL00269	Mammalian defensins proteins.	BL00269C 16.52 6.786e-26 110-139
			BL00269A 8.53 2.607e-20 45-65
			BL00269B 19.17 5.500e-17 72-101
66	BL01160	Kinesin light chain	BL01160B 19.54 8.297e-10 6-60
		repeat proteins.
67	BL00028	Zinc finger, C2H2	BL00028 16.07 5.846e-11 476-493
		type, domain proteins.	BL00028 16.07 6.192e-11 989-1006
71	BL00289	Pentaxin family proteins.	BL00289E 18.00 4.375e-13 22-37
74	BL00348	p53 tumor antigen proteins.	BL00348F 23.19 4.571e-09 140-183
75	BL00455	Putative AMP-binding	BL00455 13.31 6.684e-13 248-264
		domain proteins.
78	BL00421	Transmembrane 4	BL00421E 20.97 1.851e-09 17-47
		family proteins.
80	PD01066	PROTEIN ZINC FINGER	PD01066 19.43 2.149e-29 6-45
		ZINC-FINGER METAL-
		BINDINC NU.
81	PR00014	FIBRONECTIN TYPE III	PR00014D 12.04 2.059e-10 215-230
		REPEAT SIGNATURE
84	BL00269	Mammalian defensins proteins.	BL00269C 16.52 6.786e-26 133-162
			BL00269A 8.53 2.607e-20 68-88
			BL00269B 19.17 5.500e-17 95-124
85	PD01876	ANTIGEN MELANOMA-	PD01876C 21.73 1.231e-20 75-128
		ASSOCIATED MULTIGENE
		FAMILY TUM.
86	PD02870	RECEPTOR	PD02870B 18.83 8.835e-11 326-359
		INTERLEUKIN-1
		PRECURSOR.
87	PR00988	URIDINE KINASE	PR00988A 6.39 6.276e-12 386-376
		SIGNATURE
90	BL00120	Lipases, serine proteins.	BL00120C 12.62 9.053e-12 95-106
96	BL00027	′Homeobox′ domain proteins.	BL00027 26.43 5.500e-30 169-212
100	BL01128	Shikimate kinase proteins.	BL01128A 18.84 8.200e-14 7-41
101	PR00014	FIBRONECTIN TYPE III	PR00014C 15.44 1.783e-09 211-230
		REPEAT SIGNATURE	PR00014A 8.22 3.045e-09 373-383
			PR00014C 15.44 6.087e-09 309-328
102	DM00372	CARCINOEMBRYONIC	DM00372C 23.69 4.919e-12 67-103
		ANTIGEN PRECURSOR
		AMINO-TERMINAL
		DOMAIN.
105	BL01282	BIR repeat proteins.	BL01282B 30.49 1.000e-11 194-233
109	PR00464	E-CLASS P450 GROUP	PR00464A 20.47 9.591e-16 149-170
		II SIGNATURE	PR00464C 18.84 1.000e-15 324-353
			PR00464D 17.40 6.250e-15 353-371
			PR00464B 20.41 1.844e-12 205-224
110	PD02382	RECEPTOR CHAIN	PD02382A 17.43 9.321e-09 99-115
		PRECURSOR TRANSME.
112	BL00795	Involucrin proteins.	BL00795C 17.06 6.442e-10 905-950
115	DM00215	PROLINE-RICH PROTEIN 3.	DM00215 19.43 6.644e-09 603-636
116	BL01282	BIR repeat proteins.	BL01282B 30.49 1.000e-11 137-176
117	BL01282	BIR repeat proteins.	BL01282B 30.49 1.000e-11 187-226
118	BL00218	Amino acid permeases proteins.	BL00218D 21.49 7.324e-11 226-271
			BL00218E 23.30 3.475e-09 307-347
119	BL00994	Bacterial export	BL00994A 15.15 1.086e-09 71-118
		FHIPEP family proteins.
120	PD01066	PROTEIN ZINC FINGER	PD01066 19.43 8.385e-33 6-45
		ZINC-FINGER METAL-
		BINDING NU.
121	PD01427	TRANSFERASE	PD01427B 22.45 1.545e-11 117-158
		METHYLTRANSFERASE BI.
122	PF00168	C2 domain proteins.	PR00168C 27.49 1.750e-09 202-228
127	PR00962	LETHAL(2) GIANT	PR00962D 10.40 3.054e-10 178-202
		LARVAE PROTEIN
		SIGNATURE
130	DM01970	0 kw ZK632.12	DM01970B 8.60 2.478e-13 310-323
		YDR313C ENDOSOMAL III.
131	PF00774	Dihydropyridine sensitive	PF00774D 10.59 8.396e-09 339-365
		L-type calcium channel
		(Beta subuni.
132	BL00615	C-type lectin domain proteins.	BL00615A 16.68 3.160e-11 129-147
133	PD01066	PROTEIN ZINC FINGER	PD01066 19.43 2.705e-11 47-86
		ZINC-FINGER METAL-
		BINDING NU.
134	BL00359	Ribosomal protein	BL00359B 23.07 7.462e-24 160-201
		L11 proteins.	BL00359C 22.18 6.586e-22 215-249
			BL00359A 20.66 4.000e-21 124-160
136	PD02462	PROTEIN BOLA	PD02462A 22.48 1.220e-09 104-139
		TRANSCRIPTION
		REGULATION AC.
137	PR00679	PROHIBITIN SIGNATURE	PR00679F 8.03 6.478e-28 178-202
			PR00679C 14.44 7.677e-22 107-126
			PR00679E 12.82 5.171e-19 153-173
			PR00679D 11.91 9.053e-18 130-147
			PR00679G 6.13 7.882e-17 201-218
			PR00679B 13.63 2.444e-10 84-104
138	PR00245	OLFACTORY RECEPTOR	PR00245E 12.40 8.286e-12 45-60
		SIGNATURE
139	PD00126	PROTEIN REPEAT	PD00126A 22.53 6.885e-10 99-120
		DOMAIN TPR NUCLEA.
140	BL01145	Ribosomal protein	BL01145A 13.73 1.000e-12 3-45
		L34e proteins.
145	BL00154	E1-E2 ATPases	BL00154D 12.57 7.387e-09 95-106
		phosphorylation site proteins.
147	PD00066	PROTEIN ZINC-FINGER	PD00066 13.92 7.923e-15 439-452
		METAL-BINDI.	PD00066 13.92 2.800e-14 411-424
			PD00066 13.92 2.800e-14 467-480
			PD00066 13.92 5.800e-14 495-508
			PD00066 13.92 5.800e-14 523-536
			PD00066 13.92 8.200e-14 355-368
			PD00066 13.92 5.500e-13 579-592
			PD00066 13.92 3.143e-12 551-564
			PD00066 13.92 4.857e-12 383-396
149	BL00649	G-protein coupled	BL00649C 17.82 9.542e-12 400-426
		receptors family 2 proteins.
152	BL00479	Phorbol esters/diacylglycerol	BL00479B 12.57 8.875e-09 886-902
		binding domain proteins.
153	PR00205	CADHERIN SIGNATURE	PR00205B 11.39 5.655e-16 255-273
			PR00205A 14.73 1.000e-12 180-196
			PR00205B 11.39 4.927e-10 475-493
			PR00205C 13.65 9.438e-10 515-530
155	BL00122	Carboxylesterases	BL00122A 12.04 3.152e-15 86-107
		type-B serine proteins.	BL00122D 12.53 7.097e-14 197-213
			BL00122B 16.84 1.346e-13 148-159
			BL00122C 7.91 9.550e-10 168-179
157	BL00018	EF-hand calcium-	BL00018 7.41 2.800e-10 217-230
		binding domain proteins.	BL00018 7.41 8.650e-10 133-146
160	PD00066	PROTEIN ZINC-FINGER	PD00066 13.92 6.143e-12 927-940
		METAL-BINDI.	PD00066 13.92 7.000e-09 343-356
161	DM01857	5 kw NUCLEOSIDE	DM01857B 14.94 6.471e-19 284-312
		TRANSPORT DEPENDENT	DM01B57E 18.90 7.313e-18 488-527
		NA.	DM01857F 12.86 7.045e-15 548-575
			DM01857C 15.62 4.500e-14 312-344
			DM01857A 20.25 1.667e-13 207-250
			DM01857D 16.80 3.165e-12 372-410
164	DM01840	kw SPAC24B11.09	DM01840B 22.04 1.844e-40 59-103
		R07E5.13.	DM01840A 10.95 9.571e-13 31-43
166	BL01115	GTP-binding nuclear	BL01115A 10.22 3.438e-14 53-97
		protein ran proteins.
167	PF00622	Domain in SPla and	PF00622B 21.00 2.500e-13 265-287
		the Ryanodine Receptor.
168	PR00019	LEUCINE-RICH REPEAT	PR00019A 11.19 5.050e-11 66-80
		SIGNATURE	PR00019B 11.36 6.850e-10 63-77
169	BL00509	Ras GTPase-activating	BL00509B 10.28 5.263e-10 429-440
		proteins.
172	PR00720	MAMMALIAN LMW	PR00720C 12.41 1.099e-27 88-109
		PHOSPHOTYROSINE	PR00720B 10.61 4.789e-20 71-87
		PROTEIN PHOSPHATASE	PR00720A 16.54 2.000e-17 28-41
		SIGNATURE	PR00720E 10.01 1.342e-16 117-139
			PR00720D 17.32 1.778e-15 110-127
173	PD00131	ATP-BINDING	PD00131B 34.97 7.987e-09 108-162
		TRANSPORT
		TRANSMEMBR.
175	BL00615	C-type lectin domain	BL00615A 16.68 9.526e-13 573-591
		proteins.
179	BL00134	Serine proteases,	BL00134A 11.96 5.781e-15 493-510
		trypsin family,	BL00134B 15.99 4.194e-14 675-699
		histidine proteins.
180	BL00236	Neurotransmitter-gated	BL00236D 25.66 4.000e-30 64-106
		ion-channels proteins.
181	BL00604	Synaptophysin/	BL00604F 5.96 7.718e-10 367-412
		synaptoporin proteins.
184	PR00042	FOS TRANSFORMING	PR00042E 9.69 7.652e-09 234-258
		PROTEIN SIGNATURE
187	PD00066	PROTEIN ZINC-FINGER	PD00066 13.92 9.400e-14 365-378
		METAL-BINDI.	PD00066 13.92 6.143e-12 335-348
			PD00066 13.92 2.174e-11 395-408
188	BL00962	Ribosomal protein S2	BL00962D 22.51 5.500e-35 131-175
		proteins.	BL00962C 15.90 9.591e-17 106-124
			BL00962B 36.15 9.060e-15 40-94
189	BL00152	ATP synthase alpha	BL00152A 15.38 5.109e-14 128-154
		and beta subunits proteins.
191	BL00152	ATP synthase alpha	BL00152B 21.40 4.273e-37 124-162
		and beta subunits proteins.	BL00152A 15.38 8.364e-23 67-93
192	BL00152	ATP synthase alpha	BL00152B 21.40 2.000e-32 185-223
		and beta subunits proteins.	BL00152A 15.38 8.364e-23 128-154
193	PR00493	BREAST CANCER TYPE I	PR00493G 7.57 1.184e-10 652-673
		SUSCEPTIBILITY
		PROTEIN SIGNATURE
195	BL00301	GTP-binding	BL00301A 12.41 1.750e-12 72-84
		elongation factors proteins.
197	BL00745	Prokaryotic-type	BL00745C 13.66 7.398e-18 59-106
		class I peptide
		chain release
		factors signat.
198	BL00745	Prokaryotic-type class I peptide	BL00745C 13.66 4.706e-12 59-106
		chain release factors signat.
201	BL00660	Band 4.1 family	BL00660B 17.33 4.800e-27 136-176
		domain proteins.	BL00660A 31.50 7.911e-20 52-105
			BL00660C 23.36 2.241e-19 215-259
			BL00660E 23.41 9.647e-13 301-324
205	PR00109	TYROSINE KINASE	PR00109B 12.27 1.882e-12 155-174
		CATALYTIC DOMAIN
		SIGNATURE
207	PR00837	ALLERGEN V5/TPX-1	PR00837C 17.21 4.064e-11 155-172
		FAMILY SIGNATURE	PR00837A 14.77 4.960e-10 78-97
			PR00837B 11.64 1.310e-09 133-147
208	BL01115	GTP-binding nuclear	BL01115A 10.22 8.909e-13 4-48
		protein ran proteins.
211	BL00175	Phosphoglycerate	BL00175D 27.67 4.000e-40 367-419
		mutase famiiy	BL00175C 23.75 6.870e-28 316-348
		phosphohistidine	BL00175A 15.42 8.200e-19 252-272
		proteins.	BL00175B 12.60 8.714e-17 299-312
212	PD00066	PROTEIN ZINC-FINGER	PD00066 13.92 1.000e-14 547-560
		METAL-BINDI.	PD00066 13.92 2.200e-14 353-366
			PD00066 13.92 3.400e-14 241-254
			PD00066 13.92 6.400e-14 325-338
			PD00066 13.92 1.500e-13 297-310
			PD00066 13.92 6.500e-13 465-478
			PD00066 13.92 7.500e-13 437-450
			PD00066 13.92 8.500e-13 409-422
			PD00066 13.92 2.714e-12 269-282
			PD00066 13.92 3.571e-12 381-394
			PD00066 13.92 7.577e-10 519-532
213	PD02331	CYCLIN CELL CYCLE	PD02331C 13.84 1.913e-11 9-36
		DIVISION PROTE.
215	BL00239	Receptor tyrosine	BL00239B 25.15 3.915e-15 100-148
		kinase class II proteins.
216	BL01013	Oxysterol-binding	BL01013D 26.81 9.135e-22 501-545
		protein famiiy	BL01013A 25.14 4.600e-14 220-256
		proteins.	BL01013C 9.97 4.906e-12 330-340
			BL01013B 11.33 3.017e-11 287-298
219	BL00289	Pentaxin family protiens.	BL00289A 30.36 6.850e-26 25-56
			BL00289E 18.00 6.684e-14 78-93
220	PR00217	43 KD POSTSYNAPTIC	PR00217C 10.91 7.527e-09 547-563
		PROTEIN SIGNATRRE
221	PR00756	MEMBRANE ALANYL	PR00756D 10.58 1.529e-21 367-383
		DIPEPTIDASE (M1)	PR00756B 14.06 5.737e-16 253-269
		FAMILY SIGNATURE	PR00756A 12.90 1.237e-13 205-221
			PR00756E 11.91 4.094e-13 386-399
			PR00756C 11.60 6.108e-11 331-342
222	DM01688	2 POLY-IG RECEPTOR.	DM01688I 14.97 6.279e-09 75-123
224	PR00308	TYPE I ANTIFREEZE	PR00308C 3.83 2.523e-10 40-50
		PROTEIN SIGNATURE	PR00308C 3.83 8.892e-10 41-51
			PR00308C 3.83 8.892e-10 42-52
			PR00308B 4.28 6.671e-09 40-52
231	BL00300	SRP54-type proteins	BL00300C 25.57 6.000e-09 215-269
		GTP-binding domain proteins.
232	BL00514	Fibrinogen beta and gamma	BL00514C 17.41 9.463e-19 233-270
		chains C-terminal domain	BL00514E 14.28 7.750e-12 293-310
		proteins.	BL00514D 15.35 9.824e-11 274-287
			BL00514G 15.98 4.273e-10 356-386
			BL00514H 14.95 6.217e-09 391-416
233	BL01158	Macrophage migration	BL01158A 21.81 4.130e-30 2-47
		inhibitory factor	BL01158B 17.07 4.316e-29 47-74
		family proteins.
234	BL00456	Sodium: solute	BL00456A 22.59 6.250e-40 46-101
		symporter family	BL00456C 24.55 6.586e-40 184-239
		proteins.	BL00456B 18.94 8.125e-25 122-152
			BL00456D 6.92 5.500e-10 476-486
237	PR00830	ENDOPEPTIDASE LA (LON)	PR00830A 8.41 4.780e-14 241-261
		SERINE PROTEASE (S16)
		SIGNATURE
238	PR00165	ANION EXCHANGER	PR00165I 10.02 8.412e-14 829-849
		SIGNATURE	PR00165A 9.84 6.423e-13 495-518
			PR00165B 15.26 9.090e-11 520-541
			PR00165F 10.39 6.663e-10 639-658
239	BL00027	′Homeobox′ domain proteins.	BL00027 26.43 7.943e-14 65-108
241	BL00115	Eukaryotic RNA polymerase II	BL00115Z 3.12 2.047e-10 469-518
		heptapeptide repeat proteins.
244	BL01215	Mrp family proteins.	BL01215A 9.75 2.436e-09 466-493
245	PR00303	PREPROTEIN	PR00303G 10.45 8.759e-09 88-111
		TRANSLOCASE SECY
		SUBUNIT SIGNATURE
249	PF00429	ENV polyprotein	PF00429 31.08 8.015e-16 415-465
		(coat polyprotein).
250	BL00415	Synapsins proteins.	BL00415N 4.29 7.115e-10 224-268
252	BL00183	Ubiquitin-conjugating	BL00183 28.97 4.326e-22 81-129
		enzymes proteins.
254	BL00237	G-protein coupled	BL00237A 27.68 4.214e-16 108-148
		receptors proteins.	BL00237C 13.19 3.323e-11 245-272
			BL00237B 5.28 2.227e-09 182-194
255	BL00237	G-protein coupled	BL00237A 27.68 4.214e-16 108-148
		receptors proteins.	BL00237C 13.19 3.323e-11 280-307
			BL00237B 5.28 2.227e-09 217-229
259	PR00259	TRANSMEMBRANE FOUR	PR00259B 14.81 3.769e-21 50-77
		FAMILY SIGNATURE	PR00259C 16.40 4.000e-20 77-106
			PR00259A 9.27 3.600e-16 12-36
260	BL00282	Kazal serine protease inhibitors	BL00282 16.88 7.207e-14 562-585
		family proteins.
261	PF00922	Vesiculovirus phosphoprotein.	PF00922A 19.17 7.724e-09 88-122
266	PR00049	WILM'S TUMOUR	PR00049D 0.00 7.143e-10 17-32
		PROTEIN SIGNATURE
267	BL00612	Osteonectin domain proteins.	BL00612E 13.12 3.947e-11 379-424
268	BL00223	Annexins repeat proteins	BL00223A 15.59 1.000e-33 147-181
		domain proteins.	BL00223A 15.59 1.435e-16 75-109
			BL00223C 24.79 3.928e-15 134-189
269	BL00223	Annexins repeat proteins	BL00223B 28.47 1.000e-40 188-238
		domain proteins.	BL00223A 15.59 1.000e-33 119-153
			BL00223A 15.59 1.435e-16 47-81
			BL00223C 24.79 3.928e-15 106-161
270	BL00223	Annexins repeat proteins	BL00223A 15.59 1.000e-33 119-153
		domain proteins.	BL00223A 15.59 1.435e-16 47-81
			BL00223C 24.79 3.928e-15 106-161
273	BL00086	Cytochrome P450 cysteine	BL00086 20.87 8.615e-27 423-455
		heme-iron ligand proteins.
274	PR00385	P450 SUPERFAMILY	PR00385A 14.97 5.696e-13 295-313
		SIGNATURE
275	BL00477	Alpha-2-macroglobulin	BL00477A 13.50 9.182e-19 70-99
		family thiolester region
		proteins.
277	BL00500	Thymosin beta-4 family	BL00500 9.77 2.565e-28 2-42
		proteins.
279	PR00320	G-PROTEIN BETA WD-40	PR00320A 16.74 4.971e-10 231-246
		REPEAT SIGNATURE	PR00320C 13.01 8.200e-10 231-246
			PR00320B 12.19 9.486e-10 231-246
			PR00320B 12.19 3.475e-09 188-203
			PR00320B 12.19 4.600e-09 315-330
			PR00320C 13.01 4.900e-09 315-330
288	PF00580	UvrD/REP helicase.	PR00580D 13.15 8.920e-13 670-684
			PF00580E 13.89 2.800e-11 867-886
			PF00580F 8.62 9.438e-10 913-926
289	PR00019	LEUCINE-RICH REPEAT	PR00019B 11.36 1.000e-09 64-78
		SIGNATURE	PR00019A 11.19 8.000e-09 90-104
290	PD00126	PROTEIN REPEAT	PD00126A 22.53 5.500e-10 229-250
		DOMAIN TPR NUCLEA.
295	DM01206	CORONAVIRUS	DM01206B 10.69 4.759e-09 464-484
		NUCLEOCAPSID PROTEIN.
297	BL00972	Ubiquitin carboxyl-terminal	BL00972A 11.93 8.054e-15 191-209
		hydrolases family 2 proteins.
298	PF00023	Ank repeat proteins.	PF00023A 16.03 9.500e-12 347-363
			PF00023A 16.03 8.500e-10 283-299
			PF00023A 16.03 8.875e-10 184-200
300	BL00415	Synapsins proteins.	BL00415Q 2.23 8.297e-09 13-49
302	BL01113	C1q domain proteins.	BL01113B 18.26 2.500e-13 841-877
304	BL01248	Laminin-type EGF-like	BL01248 11.02 7.171e-12 258-271
		(LE) domain proteins.	BL01248 11.02 7.943e-12 325-338
305	BL00164	Enolase proteins.	BL00164A 11.58 2.800e-28 41-64
307	BL01153	NOL1/NOP2/sun family	BL01153D 19.69 8.322e-14 102-128
		proteins.	BL01153C 13.67 6.507e-10 51-65
308	BL00892	HIT family proteins.	BL00892B 16.86 1.000e-20 130-154
			BL00892A 18.17 6.657e-20 64-95
309	PF00791	Domain present in ZO-1 and	PF00791B 28.49 4.146e-10 73-128
		Unc5-like netrin receptors.
310	BL00615	C-type lectin domain proteins.	BL00615B 12.25 5.200e-12 166-180
311	BL00071	Glyceraldehyde 3-phosphate	BL00071B 21.70 1.000e-40 80-126
		dehydrogenase proteins.	BL00071C 11.81 1.000e-40 146-181
			BL00071D 19.39 3.118e-25 184-239
			BL00071E 11.48 4.600e-24 308-329
			BL00071A 5.81 2.607e-14 5-17
314	PR00926	MITOCHONDRIAL	PR00926F 17.75 2.688e-10 15-38
		CARRIER PROTEIN	PR00926D 10.53 6.625e-10 21-40
		SIGNATURE
315	PR00654	ANGIOTENSINOGEN	PR00654A 15.64 1.540e-26 23-44
		SIGNATURE	PR00654D 10.48 3.538e-26 153-175
			PR00654F 15.16 8.071e-26 255-275
			PR00654E 9.81 2.241e-25 194-215
			PR006540 9.50 5.500e-21 115-135
316	BL00290	Immunoglobulins and major	BL00290A 20.89 8.071e-17 34-57
		histocompatibility
		complex proteins.
317	BL00290	Immunoglobulins and major	BL00290A 20.89 7.600e-16 34-57
		histocompatibility
		complex proteins.
318	BL00290	Immunoglobulins and major	BL00290A 20.89 2.800e-16 31-54
		histocompatibility
		complex proteins.
319	BL00290	Immunoglobulins and major	BL00290A 20.89 9.400e-16 34-57
		histocompatibility
		complex proteins.
320	BL00290	Immunoglobulins and major	BL00290B 13.17 4.000e-21 282-300
		histocompatibility	BL00290A 20.89 4.600e-16 34-57
		complex proteins.	BL00290A 20.89 2.421e-13 225-248
321	BL00290	Immunoglobulins and major	BL00290A 20.89 4.600e-16 34-57
		histocompatibility
		complex proteins.
323	BL00290	Immunoglobulins and major	BL00290A 20.89 8.071e-17 34-57
		histocompatibility
		complex proteins.
324	BL00290	Immunoglobulins and major	BL00290A 20.89 4.600e-16 34-57
		histocompatibility
		complex proteins.
325	BL00290	Immunoglobulins and major	BL00290A 20.89 4.600e-16 34-57
		histocompatibility
		complex proteins.
326	PF00651	BTB (also known as BR-C/Ttk)	PF00651 15.00 2.500e-14 46-59
		domain proteins.
333	BL00972	Ubiquitin carboxyl-	BL00972A 11.93 3.919e-15 101-119
		terminal hydrolases	BL00972B 9.45 7.577e-10 180-190
		family 2 proteins.
337	PF00789	Domain present in	PF00789B 19.70 5.941e-09 213-234
		ubiquitin-regulatory proteins.
338	PF00789	Domain present in	PF00789B 19.70 5.941e-09 259-280
		ubiquitin-regulatory proteins.
339	PR00625	DNAJ PROTEIN FAMILY	PR00625A 12.84 3.000e-19 19-39
		SIGNATURE	PR00625B 13.48 2.756e-17 47-68
340	BL00518	Zinc finger, C3HC4 type	BL00518 12.23 5.714e-10 24-33
		(RING finger) proteins.
341	PR00500	POLYCYSTIC KIDNEY	PR00500I 9.22 1.107e-31 2810-2833
		DISEASE PROTEIN	PR00500G 3.68 1.087e-30 2525-2548
		SIGNATURE	PR00500H 17.80 1.107e-29 2662-2684
			PR00500E 6.99 1.106e-27 2350-2370
			PR00500F 9.44 1.108e-26 2483-2503
343	BL00415	Synapsins proteins.	BL00415N 4.29 5.401e-09 136-180
344	PD00930	PROTEIN GTPASE	PD00930B 33.72 2.800e-23 229-270
		DOMAIN ACTIVATION.	PD00930A 25.62 5.021e-12 125-151
345	BL00023	Type II fibronectin	BL00023 24.31 8.043e-34 281-318
		collagen-binding	BL00023 24.31 5.320e-32 223-260
		domain proteins.	BL00023 24.31 5.800e-29 340-377
346	BL00023	Type II fibronectin	BL00023 24.31 8.043e-34 281-318
		collagen-binding	BL00023 24.31 5.320e-32 223-260
		domain proteins.	BL00023 24.31 5.800e-29 340-377
350	DM01354	kw TRANSCRIPTASE	DM01354R 8.50 2.969e-22 2115-2145
		REVERSE II ORF2.	DM01354S 11.61 1.692e-14 2145-2166
351	BL00888	Cyclic nucleotide-	BL00888B 14.79 4.706e-18 372-396
		binding domain proteins.	BL00868A 18.03 1.000e-08 354-371
353	PF00615	Regulator of G protein	PR00615B 16.25 9.625e-16 73-90
		signalling domain proteins.	PR00615C 10.06 9.206e-12 150-164
354	BL00766	Tetrahydrofolate	BL00766E 13.78 9.625e-39 191-228
		dehydrogenase/cyclohydrolase	BL00766C 25.86 4.375e-31 77-125
		proteins.	BL00766D 17.05 5.966e-25 152-182
355	BL00180	Glutamine synthetase proteins.	BL00180E 17.60 1.000e-40 154-206
			BL00180D 13.26 2.174e-24 119-141
			BL00180F 10.05 6.211e-17 218-231
			BL00180G 10.20 8.435e-17 307-322
			BL00180C 12.14 4.600e-14 102-112
			BL00180B 18.03 4.971e-14 68-87
			BL00180A 13.20 5.065e-14 32-45
356	BL00180	Glutamine synthetase proteins.	BL00180F 10.05 6.750e-15 49-62
358	BL01131	Ribosomal RNA adenine	BL01131A 26.62 1.000e-08 77-123
		dimethylases proteins.
360	DM00191	w SPAC8A4.04C	DM00191A 8.16 5.440e-09 36-49
		RESISTANCE SPAC8A4.05C
		DAUNORUBICIN.
361	DM00191	w SPAC8A4.04C	DM00191A 8.16 5.440e-09 61-74
		RESISTANCE SPAC8A4.05C
		DAUNORUBICIN.
362	PF00606	Herpesviral Glycoprotein B.	PF00606I 20.74 7.894e-09 264-316
363	PR00209	ALPHA/BETA GLIADIN	PR00209B 4.88 9.080e-11 80-99
		FAMILY SIGNATURE	PR00209B 4.88 6.967e-10 86-105
364	PR00528	GLUCOCORTICOID	PR00528F 9.13 9.063e-09 31-51
		RECEPTOR SIGNATURE
365	PF00622	Domain in SPla and the	PF00622C 12.62 6.625e-13 759-773
		RYanodine Receptor.
366	BL00420	Speract receptor repeat	BL00420B 22.67 2.824e-25 37-92
		proteins domain proteins.	BL00420C 11.90 9.250e-12 122-133
367	PR00080	ALCOHOL	PR00080A 9.32 8.548e-10 122-134
		DEHYDROGENASE
		SUPERFAMILY
		SIGNATURE
369	BL00263	Natriuretic peptides proteins.	BL00263 11.87 5.909e-22 129-147
370	BL00609	Glycosyl hydrolases family	BL00609C 13.27 9.270e-11 249-261
		32 proteins.
371	PR00237	RHODOPSIN-LIKE GPCR	PR00237E 13.03 4.000e-10 26-50
		SUPERFAMILY
		SIGNATURE
372	BL00125	Serine/threonine specific	BL00125D 33.11 9.719e-35 23-78
		protein phosphatases proteins.
373	BL00453	FKBP-type peptidyl-prolyl	BL00453B 23.86 6.538e-26 281-315
		cis-trans isomerase proteins.	BL00453A 15.57 8.364e-12 249-264
			BL00453C 9.72 3.250e-11 323-336
374	PR00497	NEUTROPHIL CYTOSOL	PR00497A 6.92 8.261e-09 310-328
		FACTOR P40 SIGNATURE
375	PR00449	TRANSFORMING PROTEIN	PR00449A 13.20 8.269e-16 34-56
		P21 RAS SIGNATURE
376	PR00109	TYROSINE KINASE	PR00109B 12.27 9.847e-10 314-333
		CATALYTIC DOMAIN
		SIGNATURE
377	PR00109	TYROSINE KINASE	PR00109B 12.27 9.847e-10 314-333
		CATALYTIC DOMAIN
		SIGNATURE
378	BL00472	Small cytokines	BL00472C 20.76 8.225e-09 50-87
		(intercrine/chemokine)
		C—C subfamily signatur.
379	PR00704	CALPAIN CYSTEINE	PR007040 11.88 6.162e-09 132-149
		PROTEASE (C2) FAMILY
		SIGNATURE
380	PR00705	PAPAIN CYSTEINE	PR00705A 10.55 8.667e-13 155-171
		PROTEASE (C1) FAMILY	PR00705B 10.22 2.385e-10 334-345
		SIGNATURE
382	PD01270	RECEPTOR FC	PD01270A 17.22 7.443e-10 129-169
		IMMUNOGLOBULIN	PD01270A 17.22 7.387e-09 36-76
		AFFIN.
384	BL00412	Neuromodulin (GAP-43)	BL00412D 16.54 6.772e-10 250-301
		proteins.
385	BL00232	Cadherins extracellular	BL00232B 32.79 8.594e-35 151-199
		repeat proteins domain	BL00232B 32.79 5.579e-22 260-308
		proteins.	BL00232A 27.72 1.000e-20 57-90
			BL00232C 10.65 3.613e-14 258-276
			BL00232B 32.79 4.872e-11 377-425
			BL00232C 10.65 3.211e-09 480-498
387	BL00214	Cytosolic fatty-acid	BL00214B 26.51 9.000e-29 47-92
		binding proteins.	BL00214A 21.17 1.000e-24 6-32
388	PR00452	SH3 DOMAIN SIGNATURE	PR00452B 11.65 8.250e-09 509-525
390	BL00972	Ubiquitin carboxyl-terminal	BL00972A 11.93 8.759e-17 112-130
		hydrolases family 2 proteins.	BL00972D 22.55 8.116e-12 354-379
			BL00972B 9.45 7.088e-09 193-203
392	BL00243	Integrins beta chain	BL00243I 31.77 3.155e-09 1-44
		cysteine-rich domain proteins.
394	BL00476	Fatty acid desaturases family 1	BL00476F 12.75 6.551e-09 45-90
		proteins.
397	PR00320	G-PROTEIN BETA WD-40	PR00320A 16.74 9.690e-11 292-307
		REPEAT SIGNATURE	PR00320B 12.19 4.343e-10 292-307
			PR00320C 13.01 7.840e-10 292-307
402	PD01823	PROTEIN INTERGENIC	PD01823D 16.66 3.093e-15 21-42
		REGION ABC1 PRECURSOR	PD01823E 9.30 5.909e-15 75-88
		MITOCHONDRION T.
404	BL00540	Ferritin iron-binding regions	BL00540A 15.06 1.000e-40 9-50
		proteins.	BL00540B 18.82 1.000e-40 100-155
			BL00540C 13.00 7.500e-15 165-177
405	PR00294	STREPTOMYCES	PR00294A 10.44 6.444e-10 159-186
		SUBTILISIN INHIBITOR
		SIGNATURE
408	BL00428	Cell cycle proteins	BL00428A 14.30 3.613e-09 91-110
		ftsW/rodA/spoVE proteins.
411	BL00142	Neutral zinc metallopeptidases,	BL00142 8.38 7.188e-10 389-400
		zinc-binding region proteins.
412	DM00191	w SPAC8A4.04C	DM00191D 13.94 6.330e-11 232-271
		RESISTANCE	DM00191D 13.94 7.728e-11 48-87
		SPAC8A4.05C	DM00191D 13.94 5.000e-10 112-151
		DAUNORUBICIN.	DM00191D 13.94 5.667e-10 59-98
			DM00191D 13.94 5.667e-10 123-162
			DM00191D 13.94 6.583e-10 56-95
			DM00191D 13.94 8.417e-10 280-319
			DM00191D 13.94 8.917e-10 192-231
			DM00191D 13.94 1.391e-09 224-263
			DM00191D 13.94 2.409e-09 208-247
			DM00191D 13.94 4.835e-09 120-159
			DM00191D 13.94 5.304e-09 149-188
			DM00191D 13.94 5.461e-09 211-250
			DM00191D 13.94 6.322e-09 80-119
			DM00191D 13.94 7.652e-09 243-282
			DM00191D 13.94 8.513e-09 216-255
			DM00191D 13.94 9.452e-09 177-216
415	PR00023	Ank repeat proteins.	PR00023A 16.03 1.321e-09 110-126
418	PF00168	C2 domain proteins.	PR00168C 27.49 9.250e-17 320-346
419	PF00168	C2 domain proteins.	PR00168C 27.49 9.250e-17 320-346
420	BL00086	Cytochrome P450 cysteine	BL00086 20.87 1.857e-20 444-476
		heme-iron ligand proteins.
422	BL00218	Amino acid permeases	BL00218D 21.49 9.757e-11 263-308
		proteins.
423	PR00049	WILM'S TUMOUR	PR00049D 0.00 3.288e-09 35-50
		PROTEIN SIGNATURE
424	PR00380	KINESIN HEAVY CHAIN	PR00380A 14.18 4.086e-22 84-106
		SIGNATURE	PR00380C 13.18 5.286e-17 240-259
			PR00380D 9.93 7.698e-17 290-312
			PR00380B 12.64 7.805e-14 207-225
425	PR00049	WILM'S TUMOUR	PR00049D 0.00 1.915e-09 590-605
		PROTEIN SIGNATURE
426	BL00411	Kinesin motor domain	BL00411H 15.66 7.811e-22 79-110
		proteins.	BL00411G 21.39 8.683e-22 31-73
428	BL00790	Receptor tyrosine kinase	BL00790E 29.58 6.667e-12 767-815
		class V proteins.
429	BL00048	Protamine P1 proteins.	BL00048 6.39 4.038e-09 396-423
433	BL00107	Protein kinases ATP-binding	BL00107A 18.39 8.500e-27 342-373
		region proteins.
434	PR00910	LUTEOVIRUS ORF6	PR00910A 2.51 6.036e-09 48-61
		PROTEIN SIGNATURE
440	PR00019	LEUCINE-RICH REPEAT	PR00019A 11.19 7.261e-10 69-83
		SIGNATURE	PR00019B 11.36 4.600e-09 66-80
444	BL00018	EF-hand calcium-binding	BL00018 7.41 6.870e-09 42-55
		domain proteins.
448	BL00790	Receptor tyrosine	BL00790B 21.59 1.000e-40 61-113
		kinase class V proteins.	BL00790C 16.65 1.000e-40 165-219
			BL00790K 9.30 1.000e-40 657-711
			BL00790Q 15.61 1.000e-40 855-904
			BL00790O 7.68 5.929e-39 797-830
			BL00790G 22.06 5.114e-36 376-420
			BL00790R 16.20 7.469e-36 951-995
			BL00790E 29.58 7.250e-35 273-321
			BL00790J 14.21 8.200e-33 605-645
			BL00790N 13.25 1.214e-31 763-790
			BL00790I 20.01 1.931e-29 501-532
			BL00790D 12.41 2.500e-27 243-268
			BL00790H 13.42 6.478e-27 455-481
			BL00790M 8.74 8.683e-25 741-763
			BL00790P 12.33 3.755e-24 830-855
			BL00790F 15.90 5.200e-24 339-366
			BL00790L 11.16 5.909e-21 721-741
			BL00790A 19.74 1.964e-19 31-53
453	BL00027	′Homeobox′ domain proteins.	BL00027 26.43 2.000e-11 84-127
455	BL01160	Kinesin light chain	BL01160B 19.54 5.958e-09 387-441
		repeat proteins.
456	BL01113	Clq domain proteins.	BL01113B 18.26 2.500e-13 841-877
458	BL01206	Amiloride-sensitive	BL01206D 30.58 3.025e-28 363-412
		sodium channels proteins.	BL01206G 21.72 6.063e-27 530-576
			BL01206F 16.40 7.643e-15 485-506
			BL01206E 20.72 5.650e-14 427-454
			BL01206C 12.30 3.455e-12 333-352
			BL01206B 13.56 1.205e-10 313-327
460	BL01220	Phosphatidylethanolamine-	BL01220B 16.65 1.000e-40 59-100
		binding protein	BL01220C 14.75 5.846e-34 100-128
		family proteins.	BL01220A 22.62 3.400e-31 21-52
461	BL00815	Alpha-isopropylmalate and	BL00815C 21.36 3.118e-09 786-815
		homocitrate synthases proteins.
464	PR00049	WILM'S TUMOUR	PR00049D 0.00 4.051e-09 1-16
		PROTEIN SIGNATURE
470	PD00066	PROTEIN ZINC-FINGER	PD00066 13.92 5.200e-09 453-466
		METAL-BINDI.
471	BL00022	EGF-like domain proteins.	BL00022A 7.48 5.000e-09 177-184
			BL00022A 7.48 5.000e-09 241-248
			BL00022A 7.48 8.000e-09 49-56
473	PR00371	FLAVOPROTEIN	PR00371D 14.55 4.536e-11 385-405
		PYRIDINE NUCLEOTIDE
		CYTOCHROME REDUCTASE
		SIGNATURE
474	BL00028	Zinc finger, C2H2 type,	BL00028 16.07 4.462e-11 1087-1104
		domain proteins.
475	PR00260	BACTERIAL CHEMOTAXIS	PR00260C 10.26 9.294e-09 146-167
		SENSORY TRANSDUCER
		SIGNATURE
476	BL00845	CAP-Gly domain proteins.	BL00845 16.43 6.442e-21 405-430
			BL00845 16.43 9.820e-19 203-228
478	PD00066	PROTEIN ZINC-FINGER	PD00066 13.92 3.769e-15 369-382
		METAL-BINDI.	PD00066 13.92 4.462e-15 285-298
			PD00066 13.92 2.800e-14 257-270
			PD00066 13.92 5.200e-14 313-326
			PD00066 13.92 8.962e-10 341-354
481	PR00671	INHIBIN BETA B CHAIN	PR00671C 4.18 5.345e-09 9-29
		SIGNATURE
483	DM01803	1 HERPESVIRUS	DM01803A 10.51 6.855e-09 215-236
		GLYCOPROTEIN H.
484	DM01803	1 HERPESVIRUS	DM01803A 10.51 6.855e-09 251-272
		GLYCOPROTEIN H.
486	PR00545	RETINOIC ACID	PR00545A 5.35 9.430e-09 383-398
		RECEPTOR SIGNATURE
488	BL01238	GDA1/CD39 family of	BL01238A 11.72 7.840e-16 76-91
		nucleoside phosphatases
		proteins.
489	PD00930	PROTEIN GTPASE	PD00930B 33.72 2.800e-26 1256-1297
		DOMAIN ACTIVATION.	PD00930A 25.62 3.864e-13 1152-1178
493	BL00223	Annexins repeat proteins	BL00223B 28.47 1.000e-40 140-190
		domain proteins.	BL00223C 24.79 1.000e-40 217-272
			BL00223A 15.59 5.500e-32 21-55
			BL00223A 15.59 4.783e-14 230-264
			BL00223C 24.79 2.515e-10 8-63
			BL00223A 15.59 6.250e-10 71-105
496	DM01513	CAMP-DEPENDENT	DM01513A 13.61 8.568e-14 15-56
		PROTEIN KINASE
		REGULATORY CHAIN.
497	BL00603	Thymidine kinase	BL00603C 30.02 1.000e-40 152-207
		cellular-type proteins.	BL00603A 20.71 4.500e-33 63-96
			BL00603D 10.53 5.091e-18 217-232
			BL00603B 11.39 3.455e-15 132-147
498	PD01922	PROTEIN	PD01922B 21.83 7.328e-14 162-198
		PHOSPHODIESTERASE
		HYDROL.
499	BL00061	Short-chain	BL00061B 25.79 1.931e-13 99-137
		dehydrogenases/reductases
		family proteins.
500	BL01160	Kinesin light chain	BL01160B 19.54 5.958e-09 64-118
		repeat proteins.
501	PF00856	SET domain proteins.	PF00856A 26.14 8.579e-11 5-42
504	BL00030	Eukaryotic RNA-binding region	BL00030B 7.03 3.400e-10 116-126
		RNP-1 proteins.
506	PF00075	RNase H.	PF00075D 10.71 7.000e-11 517-528
			PF00075C 11.58 9.786e-11 484-496
			PF00075B 12.56 4.073e-10 449-460
			PF00075A 14.44 2.143e-09 402-419
508	BL00262	Insulin family proteins.	BL00262B 16.89 8.286e-17 68-88
			BL00262A 12.48 4.600e-15 32-50
509	PR00213	MYELIN P0 PROTEIN	PR00213E 5.51 9.775e-12 264-289
		SIGNATURE
511	BL00856	Guanylate kinase proteins.	BL00856C 29.21 2.658e-26 539-587
			BL00856B 9.61 2.946e-18 511-532
513	PR00475	HEXOKINASE FAMILY	PR00475B 14.92 6.143e-26 186-212
		SIGNATURE	PR00475E 16.08 2.742e-22 327-350
			PR00475F 9.68 4.000e-20 407-430
			PR00475A 14.06 3.118e-19 118-135
			PR00475C 11.92 6.684e-19 239-256
			PR00475G 9.08 1.692e-16 479-496
			PR00475D 13.30 2.653e-13 262-277
			PR00475G 9.08 2.650e-10 32-49
516	PR00722	CHYMOTRYPSIN SERINE	PR00722A 12.27 8.448e-14 56-72
		PROTEASE FAMILY (S1)
		SIGNATURE
518	BL00291	Prion protein.	BL00291A 4.49 9.379e-09 105-140
519	PF00534	Glycosyl transferases	PF00534B 14.47 9.581e-12 398-422
		group 1.
520	PF01105	emp24/gp25L/p24 family.	PF01105B 25.12 2.868e-25 126-178
521	PF01105	emp24/gp25L/p24 family.	PF01105B 25.12 2.868e-25 151-203
522	PR00305	14-3-3 PROTEIN ZETA	PR00305A 9.33 9.500e-36 37-67
		SIGNATURE	PR00305E 13.01 4.316e-32 177-204
			PR00305D 16.34 3.647e-30 150-177
			PR00305F 15.95 1.964e-26 204-234
			PR00305C 8.68 3.182e-26 115-138
			PR00305B 9.99 4.857e-24 84-109
			PR00305F 15.95 8.975e-15 215-245
526	PF00642	Zinc finger C-x8-C-x5-C-x3-H	PF00642 11.59 7.796e-10 676-687
		type (and similar).	PF00642 11.59 7.055e-09 276-287
527	PF00588	SpoU rRNA Methylase family.	PF00588B 17.18 8.200e-10 281-303
528	BL01160	Kinesin light chain	BL01160B 19.54 5.653e-09 791-845
		repeat proteins.
529	PR00326	GTP1/OBG GTP-BINDING	PR00326A 8.75 4.255e-14 364-385
		PROTEIN FAMILY
		SIGNATURE
531	BL01305	moaA/nifB/pqqE family	BL01305D 14.97 7.279e-09 7-22
		proteins.
532	PR00918	CALICIVIRUS NON-	PR00915A 13.76 5.807e-09 458-479
		STRUCTURAL
		POLYPROTEIN FAMILY
		SIGNATURE
533	PR00171	SUGAR TRANSPORTER	PR00171E 14.87 1.000e-08 73-86
		SIGNATURE
534	DM01418	352 FIBRILLAR	DM01418A 20.83 5.650e-23 101-149
		COLLAGEN CARBOXYL-	DM01418B 22.51 8.500e-11 166-208
		TERMINAL.	DM01418C 20.48 8.655e-10 236-278
535	DM01418	352 FIBRILLAR	DM01418A 20.83 5.650e-23 117-165
		COLLAGEN CARBOXYL-	DM01418B 22.51 8.500e-11 182-224
		TERMINAL.	DM01418C 20.48 8.655e-10 252-294
536	BL01052	Calponin family repeat	BL01052B 15.31 3.308e-11 30-56
		proteins.
538	BL00795	Involucrin proteins.	BL00795C 17.06 7.600e-09 239-284
542	BL01046	ATP-dependent serine	BL01046D 19.61 4.938e-35 452-493
		proteases, lon family,	BL01046C 17.03 9.581e-31 377-421
		serine active sit.	BL01046B 19.24 4.977e-29 331-377
543	BL00824	Elongation factor 1	BL00824B 9.21 2.338e-09 150-170
		beta/beta′/delta
		chain proteins.
547	BL00615	C-type lectin domain proteins.	BL00615A 16.68 3.880e-11 47-65
			BL00615B 12.25 2.286e-10 149-163
550	PR00119	P-TYPE CATION-	PR00119B 13.94 8.714e-12 35-50
		TRANSPORTING ATPASE	PR00119E 8.48 7.716e-11 420-440
		SUPERFAMILY
		SIGNATURE
551	BL00039	DEAD-box subfamily	BL00039D 21.67 3.455e-33 476-522
		ATP-dependent	BL00039A 18.44 8.548e-23 145-184
		helicases proteins.	BL00039C 15.63 8.500e-16 277-301
			BL00039B 19.19 1.837e-12 191-217
553	BL00115	Eukaryotic RNA polymerase II	BL00115Z 3.12 9.669e-09 116-165
		heptapeptide repeat proteins.
554	PF00168	C2 domain proteins.	PF00168B 11.83 8.000e-10 38-49
555	PP00791	Domain present in	PF00791B 28.49 4.165e-13 780-835
		ZO-1 and Unc5-like	PF00791B 28.49 6.767e-10 888-943
		netrin receptors.	PF00791C 20.98 8.059e-09 794-833
556	PF00023	Ank repeat proteins.	PF00023A 16.03 5.875e-10 437-453
			PF00023A 16.03 7.000e-10 563-579
			PF00023A 16.03 8.500e-10 248-264
			PF00023A 16.03 9.250e-10 95-111
			PF00023A 16.03 3.250e-09 596-612
			PF00023A 16.03 3.893e-09 716-732
			PF00023A 16.03 6.786e-09 62-78
			PF00023A 16.03 9.036e-09 496-512
557	BL00479	Phorbol esters/diacylglycerol	BL00479B 12.57 8.714e-09 143-159
		binding domain proteins.
558	BL00479	Phorbol esters/diacylglycerol	BL00479B 12.57 8.714e-09 167-183
		binding domain proteins.
559	BL00518	Zinc finger, C3HC4 type	BL00518 12.23 5.286e-10 239-248
		(RING finger) proteins.
562	BL00122	Carboxylesterases type-B	BL00122G 11.67 2.500e-15 15-26
		serine proteins.
563	PR00910	LUTEOVIRUS ORF6	PR00910A 2.51 1.986e-11 340-353
		PROTEIN SIGNATURE	PR00910A 2.51 1.986e-11 342-355
			PR00910A 2.51 1.986e-11 344-357
			PR00910A 2.51 9.778e-10 346-359
			PR00910A 2.51 1.107e-09 338-351
			PR00910A 2.51 3.464e-09 336-349
564	DM01970	0 kw ZK632.12	DM01970B 8.60 8.475e-15 175-188
		YDR313C ENDOSOMAL III.
566	PR00833	POLLEN ALLERGEN POA	PR00833H 2.30 8.375e-10 149-164
		PI SIGNATURE	PR00833H 2.30 2.846e-09 147-162
567	PF00992	Troponin.	PF00992A 16.67 3.368e-09 448-483
573	PR00019	LEUCINE-RICH REPEAT	PR00019A 11.19 7.333e-09 322-336
		SIGNATURE	PR00019B 11.36 9.280e-09 319-333
574	BL00315	Dehydrins proteins.	BL00315A 9.35 7.197e-10 93-121
576	PD00930	PROTEIN GTPASE	PD00930B 33.72 4.240e-16 235-276
		DOMAIN ACTIVATION.
577	BL00183	Ubiquitin-conjugating	BL00183 28.97 8.338e-14 44-92
		enzymes proteins.
579	PR00019	LEUCINE-RICH REPEAT	PR00019B 11.36 6.850e-10 132-146
		SIGNATURE	PR00019A 11.19 2.667e-09 135-149
			PR00019B 11.36 9.640e-09 180-194
			PR00019B 11.36 1.000e-08 277-291
581	BL00625	Regulator of chromosome	BL00625A 16.21 2.033e-16 567-596
		condensation (RCC1)	BL00625B 17.69 4.205e-12 561-595
		proteins.	BL00625R 17.69 9.423e-11 93-127
			BL00625B 17.69 1.444e-10 152-186
			BL00625A 16.21 1.759e-10 99-128
			BL00625A 16.21 2.739e-09 515-544
			BL00625B 17.69 3.172e-09 43-77
			BL00625A 16.21 4.170e-09 158-187
582	BL00414	Profilin proteins.	BL00414A 13.85 6.344e-13 2-16
			BL00414E 15.46 6.283e-09 121-136
583	BL00414	Profilin proteins.	BL00414A 13.85 6.344e-13 2-16
			BL00414E 15.46 6.283e-09 105-120
584	PD00301	PROTEIN REPEAT	PD00301A 10.24 8.200e-09 131-142
		MUSCLE CALCIUM-BI.
585	DM01930	2 kw FINGER SMCX	DM01930F 14.16 1.310e-27 24-60
		SMCY YDR096W.
586	BL00411	Kinesin motor domain proteins.	BL00411G 21.39 2.200e-39 77-119
			BL00411H 15.66 8.800e-33 125-156
			BL00411F 14.77 6.250e-18 33-58
588	BL00600	Aminotransferases class-III	BL00600E 16.43 5.725e-15 164-193
		pyridoxal-phosphate	BL00600G 12.43 7.000e-14 242-261
		attachment si.	BL00600F 8.77 7.480e-11 207-220
			BL00600D 8.71 1.750e-10 143-157
589	BL00838	Interleukins −4 and −13	BL00838A 12.35 8.696e-09 136-155
		proteins.
591	PR00121	SODIUM/POTASSIUM-	PR00121D 16.72 3.012e-12 261-283
		TRANSPORTING ATPASE
		SIGNATURE
592	BL00289	Pentaxin family proteins.	BL00289A 30.36 9.03le-09 331-362
594	BL01279	Protein-L-isoaspartate	BL01279A 24.27 1.000e-11 67-115
		(D-aspartate)
		O-methyltransferase signa.
597	BL00027	′Homeobox′ domain proteins.	BL00027 26.43 4.462e-32 244-287
598	PD00066	PROTEIN ZINC-FINGER	PD00066 13.92 6.400e-16 245-258
		METAL-BINDI.	PD00066 13.92 8.615e-15 329-342
			PD00066 13.92 6.000e-13 301-314
			PD00066 13.92 4.857e-12 217-230
			PD00066 13.92 1.346e-10 273-286
			PD00066 13.92 8.200e-09 357-370
599	BL00585	Ribosomal protein S5 proteins.	BL00585B 18.78 6.143e-18 303-340
			BL00585A 28.43 4.286e-16 220-272
600	PR00482	OMPTIN SERINE	PR00482C 11.02 7.968e-09 816-842
		PROTEASE SIGNATURE
601	PR00500	POLYCYSTIC KIDNEY	PR00500B 7.74 7.359e-10 56-77
		DISEASE PROTEIN
		SIGNATURE
603	PR00917	SMALL ROUND	PR00917G 10.59 8.990e-09 812-830
		STRUCTURED VIRUS
		(C37) CYSTEINE
		PROTEASE FAMILY
		SIGNATURE
605	BL00028	Zinc finger, C2H2 type,	BL00028 16.07 9.486e-09 109-126
		domain proteins.
607	PF00638	RanBP1 domain proteins.	PR00638 11.91 4.600e-18 67-82
608	BL00406	Actins proteins.	BL00406E 8.44 8.541e-28 323-373
			BL00406B 5.47 1.375e-27 82-137
			BL00406D 12.58 3.160e-26 266-321
			BL00406C 6.75 6.943e-25 141-196
			BL00406A 9.95 2.575e-20 7-42
610	BL00048	Protamine P1 proteins.	BL0004B 6.39 3.700e-09 153-180
611	PR00315	GTP-BINDING	PR00315A 11.81 5.688e-10 126-140
		ELONGATION FACTOR
		SIGNATURE
615	PF00780	Domain found in NIK1-like	PR00780B 23.03 9.908e-09 14-57
		kinases, mouse citron and
		yeast ROM.
619	BL00162	Eukaryotic-type	BL00162C 17.78 1.000e-40 88-125
		carbonic anhydrases	BL00162E 14.93 7.231e-39 171-204
		proteins.	BL00162F 22.68 5.050e-31 208-242
			BL00162A 22.92 8.714e-30 16-47
			BL00162D 15.06 7.158e-24 126-151
			BL00162B 21.43 1.375e-19 51-74
620	PR00457	ANIMAL HAEM	PR00457E 20.67 1.621e-24 414-441
		PEROXIDASE SIGNATURE	PR00457D 16.81 8.258e-21 389-410
			PR00457B 13.29 3.455e-18 223-239
			PR00457G 17.45 7.000e-18 595-616
			PR00457C 19.25 4.414e-16 371-390
			PR00457H 15.90 8.650e-14 666-681
			PR00457A 15.80 5.645e-12 169-181
			PR00457F 13.69 8.875e-11 467-478
621	BL01304	ubiH/COQ6 monooxygenase	BL01304A 8.05 3.571e-11 50-64
		family proteins.
623	BL00019	Actinin-type actin-binding	BL00019D 15.33 3.880e-17 145-175
		domain proteins.
625	BL00893	mutT domain proteins.	BL00893 18.99 5.500e-16 127-152
626	PF00632	HECT-domain	PF00632B 18.45 7.000e-16 488-516
		(ubiquitin-transferase).	PF00632C 20.66 7.851e-14 533-565
628	PR00239	MOLLUSCAN RHODOPSIN	PR00239E 1.58 9.566e-10 292-304
		C-TERMINAL TAIL
		SIGNATURE
634	BL01280	Glucose inhibited division	BL01280A 15.97 6.727e-36 69-110
		protein A family proteins.	BL01280B 23.56 8.105e-27 128-180
638	PR00413	HALOACID	PR00413E 15.78 6.714e-09 70-87
		DEHALOGENASE/EPOXIDE
		HYDROLASE FAMILY
		SIGNATURE
639	PD01861	PROTEIN NUCLEAR	PD01861A 14.06 6.318e-10 60-84
		RIBONUCLEOPROTEIN
		SMALL MRNA RNA.
640	PD00289	PROTEIN SH3 DOMAIN	PD00289 9.97 6.586e-09 46-60
		REPEAT PRESYNA.
643	BL00914	Syntaxin/	BL00914 24.91 1.250e-29 184-234
		epimorphin family proteins.
644	PR00010	TYPE II EGF-LIKE	PR00010C 11.16 6.667e-11 363-374
		SIGNATURE
645	PR00625	DNAJ PROTEIN FAMILY	PR00625A 12.84 4.600e-20 14-34
		SIGNATURE	PR00625B 13.48 8.759e-20 46-67
646	PD02448	TRANSCRIPTION	PD02448A 9.37 3.854e-09 351-390
		PROTEIN DNA-BINDIN.
650	PD02446	TRANSCRIPTION	PD02448A 9.37 1.511e-20 50-89
		PROTEIN DNA-BINDIN.	PD02448B 10.17 8.071e-19 89-137
654	PR00403	WW DOMAIN SIGNATURE	PR00403B 12.19 9.816e-11 144-159
			PR00403B 12.19 8.167e-10 103-118
655	PR00403	WW DOMAIN SIGNATURE	PR00403B 12.19 9.816e-11 107-122
657	PR00929	AT-HOOK-LIKE DOMAIN	PR00929B 4.38 4.600e-10 358-370
		SIGNATURE
658	PD02379	AMINOTRANSFERASE	PD02379E 11.43 1.000e-40 194-236
		BIOSYNTHESIS	PD02379F 18.62 6.029e-35 245-284
		PHOSPHOSERINE SER.	PD02379H 16.03 5.235e-33 352-385
			PD02379B 12.05 3.613e-31 80-113
			PD02379A 15.57 2.800e-25 29-60
			PD023790 13.34 3.700e-21 119-139
			PD02379D 11.83 9.419e-16 168-181
			PD02379G 10.62 2.537e-14 313-328
659	PD02379	AMINOTRANSFERASE	PD02379E 11.43 1.000e-40 194-236
		BIOSYNTHESIS	PD02379F 18.62 6.029e-35 245-284
		PHOSPHOSERINE SER.	PD02379B 12.05 3.613e-31 80-113
			PD02379A 15.57 2.800e-25 29-60
			PD02379H 16.03 7.864e-23 306-339
			PD023790 13.34 3.700e-21 119-139
			PD02379D 11.83 9.419e-16 168-181
662	PR00874	FUNGI-IV	PR00874C 4.37 6.625e-09 33-48
		METALLOTHIONEIN
		SIGNATURE
666	BL00035	′POU′ domain proteins.	BL00035B 14.46 6.236e-09 683-704
668	PF00168	C2 domain proteins.	PF00168C 27.49 8.412e-13 634-660
669	PR00168	C2 domain proteins.	PR00168C 27.49 8.412e-13 115-141
671	PR00566	DOPAMINE 1B RECEPTOR	PR00566E 13.44 5.255e-18 466-483
		SIGNATURE	PR00566A 9.32 3.000e-17 200-214
			PR00566D 9.35 1.600e-12 446-455
			PR00566C 11.44 2.184e-12 401-412
			PR00566B 8.20 3.053e-11 341-351
672	PR00169	POTASSIUN CHANNEL	PR00169A 16.77 7.851e-11 46-66
		SIGNATURE
673	DM00215	PROLINE-RICH PROTEIN 3.	DM00215 19.43 7.529e-11 183-216
674	BL00951	ER lumen protein retaining	BL00951B 14.23 1.670e-09 43-74
		receptor proteins.
675	BL00292	Cyclins proteins.	BL00292B 20.31 3.925e-11 120-151
676	PR00048	C2H2-TYPE ZINC	PR00048A 10.52 3.160e-09 111-125
		FINGER SIGNATURE
677	BL01226	Hydroxymethylglutary	BL01226I 25.06 8.560e-09 256-304
		1-coenzyme A
		synthase proteins.
681	BL00030	Eukaryotic RNA-binding region	BL00030A 14.39 1.563e-12 72-91
		RNP-1 proteins.	BL00030A 14.39 2.125e-12 156-175
689	BL00740	MAM domain proteins.	BL00740B 19.76 3.813e-09 637-658
693	BL00027	′Homeobox′ domain proteins.	BL00027 26.43 7.000e-11 93-136
698	PR00259	TRANSMEMBRANE FOUR	PR00259A 9.27 3.308e-18 19-43
		FAMILY SIGNATURE	PR00259C 16.40 9.800e-18 88-117
			PR00259D 13.50 2.756e-15 238-265
703	PR00669	INHIBIN ALPHA CHAIN	PR00669F 5.57 9.899e-09 223-241
		SIGNATURE
704	DM01292	ESICULAR LUMEN	DM01292L 12.54 9.505e-09 240-265
		DOMAIN.
705	PR00128	COLIPASE SIGNATURE	PR00128D 9.77 6.250e-25 47-66
			PR00128C 9.28 5.299e-20 24-47
708	PR00049	WILM'S TUMOUR	PR00049D 0.00 9.929e-10 384-399
		PROTEIN SIGNATURE
710	BL01118	Translation initiation	BL01118B 26.75 8.579e-26 94-132
		factor SUI1 proteins.	BL01118A 12.46 4.000e-13 77-92
711	BL00811	Oleosins proteins.	BL00811A 8.26 3.310e-09 120-158
712	BL00674	AAA-protein family proteins.	BL00674B 4.46 9.182e-11 184-206
713	DM01871	kw SSR LIGASE	DM018710 20.79 9.836e-10 270-296
		CYCLOFORMYL-
		TETRAHYDROFOLATE.
715	PR00049	WILM'S TUMOUR	PR00049D 0.00 7.712e-09 95-110
		PROTEIN SIGNATURE
717	BL01181	Ribosomal protein	BL01181 15.43 2.500e-10 13-49
		S21 proteins.
718	PR00259	TRANSMEMBRANE FOUR	PR002590 16.40 6.824e-16 88-117
		FAMILY SIGNATURE	PR00259A 9.27 3.423e-14 24-48
			PR00259D 13.50 1.574e-13 238-265
			PR00259B 14.81 8.714e-13 61-88
720	PR00496	NAPIN SIGNATURE	PR00496A 6.68 6.276e-09 21-43
723	PR00237	RHODOPSIN-LIKE GPCR	PR00237G 19.63 2.543e-11 670-697
		SUPERFAMILY	PR00237A 11.48 3.000e-10 424-449
		SIGNATURE
724	BL00972	Ubiquitin carboxyl-terminal	BL00972A 11.93 7.500e-20 36-54
		terminal hydrolases	BL00972D 22.55 6.806e-16 296-321
		family 2 proteins.	BL00972B 9.45 1.000e-13 116-126
			BL00972E 20.72 8.773e-12 321-343
725	PF00646	F-box domain proteins.	PF00646A 14.37 6.906e-09 92-106
727	BL00933	FGGY family of	BL00933D 24.01 7.545e-15 212-249
		carbohydrate kinases	BL00933B 15.94 2.200e-09 54-65
		proteins.	BL00933E 13.80 3.543e-09 439-455
			BL00933A 17.50 4.857e-09 20-44
728	PR00876	NEMATODE	PR00876B 7.66 1.887e-10 137-151
		METALLOTHIONEIN
		SIGNATURE
731	PD01976	KINASE DEHYDROGENASE	PD01976A 8.95 1.493e-09 83-96
		TRANSFERASE.
732	BL00623	GMC oxidoreductases proteins.	BL00623A 12.60 9.859e-10 12-31
735	BL01172	Ribosomal protein	BL01172B 14.10 8.909e-38 15-57
		L44e proteins.	BL01172C 16.78 7.188e-31 63-102
737	DM01724	kw ALLERGEN POLLEN	DM01724 8.14 5.909e-11 11-31
		CIM1 HOL-LI.	DM01724 8.14 6.591e-11 41-61
			DM01724 8.14 6.831e-10 39-59
			DM01724 8.14 8.697e-09 55-75
738	PR00320	G-PROTEIN BETA WD-40	PR00320A 16.74 8.463e-09 73-88
		REPEAT SIGNATURE
742	PD02269	CYTIDINE DEAMINASE	PD022690 16.36 7.882e-17 79-92
		HYDROLASE ZINC	PD02269A 10.06 1.000e-15 29-41
		AMINOHY.	PD02269D 11.98 5.000e-14 110-125
743	BL00790	Receptor tyrosine	BL00790D 12.41 8.297e-09 429-454
		kinase class V proteins.
744	BL00750	Chaperonins TCP-1 proteins.	BL00750B 16.17 2.000e-39 69-119
			BL00750A 20.07 8.286e-36 25-68
			BL00750C 25.65 8.579e-23 152-184
746	BL00415	Synapsins proteins.	BL00415N 4.29 4.710e-10 225-269
748	BL00028	Zinc finger, C2H2 type	BL00028 16.07 1.000e-09 212-229
		domain proteins.	BL00028 16.07 6.143e-09 365-382
752	BL00030	Eukaryotic RNA-binding region	BL00030A 14.39 6.143e-13 332-351
		RNP-1 proteins.
753	PF00023	Ank repeat proteins.	PR00023A 16.03 8.500e-10 283-299
			PF00023A 16.03 9.625e-10 347-363
			PF00023A 16.03 1.321e-09 184-200
			PF00023A 16.03 1.643e-09 150-166
754	BL00107	Protein kinases ATP-binding	BL00107B 13.31 1.643e-10 202-218
		region proteins.
760	PR00671	INHIBIN BETA B CHAIN	PR006710 4.18 8.966e-09 212-232
		SIGNATURE
761	PR00678	P13 KINASE P85	PR00678H 9.13 7.805e-12 292-315
		REGULATORY SUBUNIT
		SIGNATURE
762	PR00412	EPOXIDE HYDROLASE	PR00412C 11.30 2.421e-12 169-183
		SIGNATURE	PR00412A 13.23 7.947e-12 104-123
			PR00412B 12.59 7.429e-10 123-139
763	PR00217	43 KD POSTSYNAPTIC	PR00217C 10.91 7.247e-10 293-309
		PROTEIN SIGNATURE
764	PR00320	G-PROTEIN BETA WD-40	PR00320A 16.74 9.122e-09 277-292
		REPEAT SIGNATURE	PR00320A 16.74 9.780e-09 233-248
			PR00320C 13.01 1.000e-08 233-248
765	BL01230	RNA methyltransferase	BL01230E 15.79 2.918e-11 487-503
		trmA family proteins.
771	BL00039	DEAD-box subfamily	BL00039D 21.67 2.957e-09 434-480
		ATP-dependent
		helicases proteins.
773	PR00453	VON WILLEBRAND	PR00453A 12.79 2.957e-10 33-51
		FACTOR TYPE A DOMAIN
		SIGNATURE
777	PR00493	BREAST CANCER TYPE I	PR00493G 7.57 3.711e-14 693-714
		SUSCEPTIBILITY
		PROTEIN SIGNATURE
778	BL00443	Glutamine amidotransferases	BL00443F 16.68 8.714e-09 85-101
		class-II proteins.
780	DM01206	CORONAVIRUS	DM01206B 10.69 7.288e-10 167-187
		NUCLEOCAPSID
		PROTEIN.
783	BL01221	PMP-22/EMP/MP20	BL012210 26.20 1.281e-34 59-104
		family proteins.	BL01221D 13.99 5.966e-27 136-163
			BL01221A 17.26 2.385e-26 1-29
			BL01221B 13.29 1.000e-14 38-52
785	BL00027	′Homeobox′ domain proteins.	BL00027 26.43 4.000e-10 297-340
786	BL00027	′Homeobox′ domain proteins.	BL00027 26.43 4.000e-10 297-340
790	BL00303	S-100/ICaBP type calcium	BL00303B 26.15 5.075e-13 73-110
		binding protein.
792	PD01941	TRANSMEMBRANE	PD01941C 19.96 4.960e-16 84-139
		COTRANSPORTER SYMP.	PD01941B 15.02 2.093e-11 4-51
794	BL00672	Serine proteases, V8 family	BL00672B 9.84 3.554e-09 214-231
		histidine proteins.
797	BL00674	AAA-protein family proteins.	BL00674B 4.46 7.814e-10 360-382
798	BL00674	AAA-protein family proteins.	BL00674B 4.46 7.814e-10 360-382
799	BL00215	Mitochondrial energy	BL00215A 15.82 9.591e-16 206-231
		transfer proteins.	BL00215A 15.82 4.000e-15 104-129
			BL00215A 15.82 9.400e-15 7-32
			BL00215B 10.44 1.000e-10 154-167
801	BL01013	Oxysterol-binding protein	BL01013A 25.14 5.500e-21 537-573
		family proteins.	BL01013D 26.81 2.16le-18 807-851
			BL01013C 9.97 4.231e-13 625-635
			BL01013B 11.33 3.017e-11 603-614
803	BL00711	Lipoxygenases iron-binding	BL00711I 18.56 8.630e-28 577-615
		binding region proteins.	BL00711E 19.66 3.550e-22 414-451
			BL00711G 21.83 9.100e-22 503-535
			BL00711C 20.75 5.959e-19 268-297
			BL00711D 17.56 1.923e-16 347-373
			BL00711H 23.34 1.771e-12 535-574
			BL00711F 19.79 2.086e-10 484-501
805	PR00492	RHO PROTEIN GDP	PR00492C 9.68 1.900e-23 122-139
		DISSOCIATION	PR00492B 9.77 8.579e-23 76-95
		INHIBITOR SIGNATURE	PR004920 14.82 8.200e-21 139-155
			PR00492A 11.92 1.643e-18 60-76
808	BL00378	Hexokinases proteins.	BL00378A 19.01 8.500e-09 403-431
809	BL00027	′Homeobox′ domain proteins.	BL00027 26.43 8.615e-33 35-78
810	PR00179	LIPOCALIN SIGNATURE	PR00179B 9.56 1.000e-12 102-115
			PR001790 19.02 1.000e-10 130-146
			PR00179A 13.78 5.680e-10 37-50
811	BL00290	Immunoglobulins and major	BL00290A 20.89 1.818e-11 164-187
		histocompatibility
		complex proteins.
814	BL00269	Mammalian defensins proteins.	BL002690 16.52 7.158e-09 171-200
815	BL00216	Sugar transport proteins.	BL00216B 27.64 5.846e-09 141-191
818	BL00456	Sodium:solute symporter	BL00456A 22.59 2.080e-30 83-138
		family proteins.	BL00456C 24.55 3.721e-29 221-276
			BL00456B 18.94 1.000e-22 159-189
819	BL00142	Neutral zinc metallopeptidases,	BL00142 8.38 1.857e-09 494-505
		zinc-binding region proteins.
820	BL00509	Ras GTPase-activating	BL00509B 10.28 1.643e-12 610-621
		proteins.
824	PR00048	C2H2-TYPE ZINC	PR00048A 10.52 6.143e-13 252-266
		FINGER SIGNATURE	PR00048A 10.52 7.429e-13 476-490
			PR00048A 10.52 3.118e-12 336-350
			PR00048A 10.52 3.118e-12 364-378
			PR00048A 10.52 4.706e-12 504-518
			PR00048A 10.52 8.412e-12 224-238
			PR00048A 10.52 3.842e-11 392-406
			PR00046A 10.52 6.211e-11 308-322
			PR00048A 10.52 6.211e-11 448-462
			PR00048B 6.02 7.231e-11 492-502
			PR00048B 6.02 3.250e-10 240-250
			PR00048A 10.52 6.870e-10 420-434
			PR00048B 6.02 2.421e-09 380-390
825	PR00122	VACUOLAR ATP	PR00122D 9.97 7.214e-11 103-127
		SYNTRASE 16 KD	PR00122C 8.20 9.526e-10 76-103
		SUBUNIT SIGNATURE
826	BL00518	Zinc finger, C3HC4 type	BL00518 12.23 6.571e-10 30-39
		(RING finger), proteins.
828	BL01160	Kinesin light chain	BL01160B 19.54 1.610e-09 33-87
		repeat proteins.	BL01160B 19.54 9.619e-09 65-119
835	PD02411	PROTEIN	PD02411 21.89 6.786e-15 3967-4001
		TRANSCRIPTION
		REGULATION NUCLEAR.
836	DM01970	0 kw ZK632.12	DM01970B 8.60 9.423e-10 111-124
		YDR313C ENDOSOMAL III.
837	PR00048	C2H2-TYPE ZINC	PR00048A 10.52 2.174e-10 177-191
		FINGER SIGNATURE
842	PF00922	Vesiculovirus phosphoprotein.	PF00922A 19.17 7.724e-09 276-310
844	PD02059	CORE POLYPROTEIN	PD02059A 28.10 5.950e-10 34-75
		PROTEIN GAG
		CONTAINS: P.
846	BL00326	Tropomyosins proteins.	BL00326D 8.76 8.065e-09 165-206
847	BL00326	Tropomyosins proteins.	BL00326D 8.76 8.065e-09 173-214
849	PR00563	BETA-3 ADRENERGIC	PR00563B 3.98 8.141e-09 8-28
		RECEPTOR SIGNATURE
851	PR00450	RECOVERIN FAMILY	PR00450C 12.22 1.570e-09 285-307
		SIGNATURE
853	PD00066	PROTEIN ZINC-FINGER	PD00066 13.92 8.800e-14 290-303
		METAL-BINDI.	PD00066 13.92 4.000e-13 234-247
			PD00066 13.92 4.429e-12 262-275
			PD00066 13.92 9.217e-11 206-219
			PD00066 13.92 3.769e-10 505-518
			PD00066 13.92 4.115e-10 449-462
			PD00066 13.92 4.462e-10 533-546
			PD00066 13.92 6.538e-10 477-490
854	BL00615	C-type lectin domain proteins.	BL00615A 16.68 8.920e-11 137-155
855	BL00615	C-type lectin domain proteins.	BL00615A 16.68 8.920e-11 176-194
856	BL00018	EF-hand calcium-binding	BL00018 7.41 9.000e-14 65-78
		domain proteins.
858	PR00019	LEUCINE-RICH REPEAT	PR00019B 11.36 1.000e-09 219-233
		SIGNATURE
860	PD02474	SYNTHASE SMALL	PD02474B 21.08 8.568e-09 199-238
		SUBUNIT ACETOLACT.
861	PF00922	Vesiculovirus phosphoprotein.	PF00922A 19.17 1.000e-08 249-283
864	PR00289	DISINTEGRIN	PR00289B 11.79 1.947e-09 522-535
		SIGNATURE
866	PF00242	DNA polymerase (viral)	PF00242F 12.18 8.522e-09 197-219
		N-terminal domain proteins.
867	PR00780	LEUSERPIN 2	PR00780B 4.89 4.491e-09 262-285
		SIGNATURE
868	BL00226	Intermediate filaments proteins.	BL00226D 19.10 8.027e-13 208-255
869	PD01876	ANTIGEN MELANOMA-	PD01876C 21.73 3.326e-15 461-534
		ASSOCIATED MULTIGENE	PD01876C 21.73 3.045e-10 735-788
		FAMILY TUM.
870	PR00747	GLYCOSYL HYDROLASE	PR00747C 12.06 8.767e-09 337-356
		FAMILY 47 SIGNATURE
872	DM01782	HYDROGENASE (FE)	DM01782C 13.88 4.400e-19 349-368
		LARGE CHAIN.	DM01782F 9.01 4.375e-18 499-515
			DM01782B 17.29 3.412e-10 294-327
873	BL00226	Intermediate filaments proteins.	BL00226D 19.10 7.375e-38 321-368
			BL00226B 23.86 7.107e-32 155-203
			BL00226C 13.23 3.100e-19 220-251
			BL00226A 12.77 7.000e-15 55-70
			BL00226D 19.10 7.800e-09 254-301
874	DM01415	6 SALIVARY GLUE	DM01415B 13.78 9.518e-10 4-52
		PROTEIN.
876	PR00860	VERTEBRATE	PR00860B 7.04 2.929e-20 74-88
		METALLOTHIONEIN	PR00860A 5.46 5.655e-13 52-65
		SIGNATURE	PR00860C 9.61 2.400e-12 88-98
877	PR00360	C2 DOMAIN SIGNATURE	PR00360B 13.61 7.136e-09 572-586
881	DM01206	CORONAVIRUS	DM01206B 10.69 8.767e-10 567-587
		NUCLEOCAPSID	DM01206B 10.69 1.000e-09 563-583
		PROTEIN.
882	BL00092	N-6 Adenine-specific DNA	BL00092 5.35 2.000e-09 136-145
		methylases proteins.
883	PR00511	TEKTIN SIGNATURE	PR00511A 13.59 3.700e-14 113-130
885	PR00764	COMPLEMENT C9	PR00764F 16.69 2.286e-09 158-179
		SIGNATURE
887	BL01279	Protein-L-isoaspartate	BL01279A 24.27 3.691e-09 419-467
		(D-aspartate)
		O-methyltransferase signa.
889	PD01719	PRECURSOR	PD01719A 12.89 2.603e-11 259-287
		GLYCOPROTEIN SIGNAL	PD01719A 12.89 8.105e-10 199-227
		RE.
890	BL01162	Quinone oxidoreductase/	BL01162C 22.80 1.269e-18 151-195
		zeta-crystallin proteins.	BL01162A 15.38 1.265e-11 64-87
893	PD01066	PROTEIN ZINC FINGER	PD01066 19.43 5.415e-26 46-85
		ZINC-FINGER METAL-
		BINDING NU.
894	PD02910	TRANSCRIPTION	PD02910A 15.43 9.839e-09 62-97
		PROTEIN FACTOR
		REGULATION A.
895	PD02199	SUBUNIT HYDROGEN ION	PD02199A 20.58 1.000e-40 10-61
		TRANSPORT T.	PD02199D 13.18 1.000e-40 364-405
			PD02199F 15.02 1.000e-40 440-482
			PD02199J 11.42 1.000e-40 723-762
			PD02199K 15.22 1.000e-40 792-831
			PD02199G 9.43 4.447e-24 531-555
			PD02199B 27.90 1.474e-22 263-306
			PD02199H 13.62 2.636e-21 576-599
			PD02199E 7.56 8.642e-19 405-424
			PD02199C 17.60 8.085e-14 313-329
			PD02199I 8.90 4.780e-09 616-624
896	BL00218	Amino acid permeases proteins.	BL00218E 23.30 5.920e-10 343-383
897	BL00048	Protamine P1 proteins.	BL00048 6.39 9.526e-10 160-187
900	BL00811	Oleosins proteins.	BL00811B 10.57 9.791e-09 307-336
904	BL00415	Synapsins proteins.	BL00415N 4.29 4.153e-09 301-345
905	BL00107	Protein kinases ATP-binding	BL00107A 18.39 3.250e-17 133-164
		region proteins.
906	PR00449	TRANSFORMING PROTEIN	PR00449A 13.20 4.971e-14 4-26
		P21 RAS SIGNATURE
908	BL00317	WAP-type ‘four-disulfide core’	BL00317B 14.58 3.550e-13 48-70
		domain proteins.
909	BL01019	ADP-ribosylation factors	BL01019B 19.49 7.517e-21 95-150
		family proteins.
910	PD00066	PROTEIN ZINC-FINGER	PD00066 13.92 2.385e-15 128-141
		METAL-BINDI.	PD00066 13.92 5.714e-12 100-113
911	PR00048	C2H2-TYPE ZINC	PR00045A 10.52 1.000e-11 174-188
		FINGER SIGNATURE	PR00048B 6.02 1.692e-11 162-172
912	PF00651	BTB (also known as BR-C/Ttk)	PR00651 15.00 2.895e-11 45-58
		domain proteins.
913	DM00547	1 kw CHROMO	DM00547F 23.43 7.643e-34 606-653
		BROMODOMAIN SHADOW	DM00547B 11.28 7.907e-16 155-169
		GLOBAL.	DM00547C 17.30 8.650e-14 209-231
			DM00547D 11.60 6.500e-13 277-291
			DM00547E 13.94 1.000e-11 307-330
914	BL01115	GTP-binding nuclear protein	BL01115A 10.22 5.330e-11 18-62
		ran proteins.
919	BL01283	T-box domain proteins.	BL01283D 11.70 7.868e-31 59-92
			BL01283C 13.05 2.537e-14 25-39
922	DM01503	1 HERPESVIRUS	DM01803A 10.51 8.699e-09 100-121
		GLYCOPROTEIN H.
924	BL00470	Isocitrate and isopropylmalate	BL00470A 16.25 5.179e-14 10-31
		dehydrogenases proteins.	BL00470C 15.43 4.103e-10 223-238
			BL00470E 16.52 1.900e-09 287-297
925	PF00023	Ank repeat proteins.	PF00023A 16.03 3.893e-09 44-60
			PF00023B 14.20 9.182e-09 40-50
926	PF00023	Ank repeat proteins.	PF00023A 16.03 3.893e-09 72-88
			PF00023B 14.20 9.182e-09 68-78
929	PD01066	PROTEIN ZINC FINGER	PD01066 19.43 2.019e-26 51-90
		ZINC-FINGER METAL-
		BINDING NU.
930	PR00830	ENDOPEPTIDASE LA	PR00830A 8.41 4.927e-13 222-242
		(LON) SERINE
		PROTEASE (S16)
		SIGNATURE
931	BL00456	Sodium:solute symporter	BL00456A 22.59 1.957e-32 35-90
		family proteins.	BL00456B 18.94 9.780e-17 111-141
932	BL00456	Sodium:solute symporter	BL00456A 22.59 1.957e-32 35-90
		family proteins.	BL00456C 24.55 1.225e-31 173-228
			BL00456B 18.94 9.780e-17 111-141
936	BL00290	Immunoglobulins and major	BL00290A 20.89 1.818e-11 159-182
		histocompatibility
		complex proteins.
937	PR00830	ENDOPEPTIDASE LA	PR0083CA 8.41 5.897e-10 352-372
		(LON) SERINE
		PROTEASE (S16)
		SIGNATURE
938	PF00651	BTB (also known as BR-C/TtK)	PR00651 15.00 7.000e-10 50-63
		domain proteins.
939	PD00306	PROTEIN GLYCOPROTEIN	PD00306A 10.26 6.625e-13 544-558
		PRECURSOR RE.
940	PD00306	PROTEIN GLYCOPROTEIN	PD00306A 10.26 6.625e-13 544-558
		PRECURSOR RE.
941	PD00306	PROTEIN GLYCOPROTEIN	PD00306A 10.26 6.625e-13 544-558
		PRECURSOR RE.
942	PF00938	Lipoprotein.	PF00938E 19.50 6.096e-09 272-307
943	PF00925	GTP cyclohydrolase II.	PF00925F 13.23 9.850e-09 356-367
945	BL00226	Intermediate filaments proteins.	BL00226A 12.77 5.355e-13 139-154
947	PF00035	Double-stranded RNA	PF00035B 12.06 7.750e-09 273-287
		binding motif.
948	PF00622	Domain in SPla and the	PF00622B 21.00 9.250e-11 170-192
		RYanodine Receptor.
*Results include in order: accession number subtype; raw score; p-value; position of signature in amino acid sequence.

TABLE 4
SEQ
ID NO:	pFAM NAME	DESCRIPTION	p-value	pFAM SCORE
2	UCH-2	Ubiquitin carboxyl-terminal hydrolase family 2	5.9e-26	99.7
5	SH2	Src homology domain 2	8.5e-22	66.5
6	SH2	Src homology domain 2	8.5e-22	66.5
7	RCC1	Regulator of chromosome condensation (RCC1)	4.4e-18	68.4
14	UQ_con	Ubiquitin-conjugating enzyme	3.2e-49	176.9
15	UQ_con	Ubiquitin-conjugating enzyme	1.2e-68	241.5
20	gntR	Bacterial regulatory proteins, gntR family	0.062	11.4
21	gntR	Bacterial regulatory proteins, gntR family	0.062	11.4
23	kinesin	Kinesin motor domain	2.9e-128	439.5
27	Kelch	Kelch motif	7.9e-71	248.7
29	kinesin	Kinesin motor domain	1.9e-171	583.0
30	pkinase	Eukaryotic protein kinase domain	8.5e-09	35.7
32	Aa_trans	Transmembrane amino acid transporter protein	3.1e-53	190.3
34	EGF	EGF-like domain	3.9e-24	93.6
38	ig	Immunoglobulin domain	6.2e-06	24.2
39	LRR	Leucine Rich Repeat	1.1e-10	48.9
43	CK_II_beta	Casein kinase II regulatory subunit	4.5e-69	242.9
45	COX6B	Cytochrome oxidase c subunit VIb	0.064	−6.7
46	RNA_pol_B	RNA polymerase beta subunit	0	1206.8
47	zf-C3HC4	Zinc finger, C3HC4 type (RING finger)	2e-07	28.3
48	HLH	Helix-loop-helix DNA-binding domain	1.5e-07	38.5
49	ras	Ras family	3.3e-12	2.0
50	pkinase	Eukaryotic protein kinase domain	1.9e-43	157.8
52	ig	Immunoglobulin domain	7.2e-08	30.4
54	PX	PX domain	3.7e-06	33.9
65	Defensin_propep	Defensin propeptide	3e-25	97.3
67	zf-C2H2	Zinc finger, C2H2 type	2e-55	197.5
71	pentaxin	Pentaxin family	3.4e-18	66.3
73	SSF	Sodium:solute symporter family	1.7e-05	−65.8
75	AMP-binding	AMP-binding enzyme	1.1e-12	−49.0
78	transmembrane4	Transmembrane 4 family	6.4e-05	18.7
80	zf-C2H2	Zinc finger, C2H2 type	4.9e-30	113.2
81	fn3	Fibronectin type III domain	1.8e-13	58.2
83	polyprenyl_synt	Polyprenyl synthetases	0.015	−83.6
84	Defensin_propep	Defensin propeptide	3e-25	97.3
85	MAGE	MAGE family	3.5e-34	127.0
86	LRR	Leucine Rich Repeat	7.7e-15	62.7
87	Cytidylyltransf	Cytidylyltransferase	1.4e-05	29.3
90	lipase	Lipase	2.5e-15	55.2
96	homeobox	Homeobox domain	1.1e-30	115.3
101	fn3	Fibronectin type III domain	4.7e-78	272.7
102	ig	Immunoglobulin domain	0.00045	18.2
103	MHC_I	Class I Histocompatibility antigen, domains alpha 1 and 2	6.7e-08	32.8
104	Trans_recep	Transient receptor	1.9e-34	115.7
105	Tropomyosin	Tropomyosins	0.0086	11.3
109	p450	Cytochrome P450	1.8e-61	217.6
116	Tropomyosin	Tropomyosins	0.0086	11.3
117	Tropomyosin	Tropomyosins	0.0086	11.3
118	aa_permeases	Amino acid permease	3.2e-06	−173.3
120	zf-C2H2	Zinc finger, C2H2 type	1.3e-124	427.4
122	C2	C2 domain	1.1e-38	142.0
123	ig	Immunoglobulin domain	0.00079	17.4
127	WD40	WD domain, G-beta repeat	9.6e-15	62.4
130	FYVE	FYVE zinc finger	7.4e-23	86.0
131	PH	PH domain	3.9e-25	94.8
133	KRAB	KRAB box	1.6e-24	94.9
134	Ribosomal_L11	Ribosomal protein L11	3.9e-64	226.5
136	zf-C2H2	Zinc finger, C2H2 type	2.1e-185	629.4
137	Band_7	SPFH domain/Band 7 family	6.5e-35	129.4
139	TPR	TPR Domain	5.3e-16	66.6
142	WH1	WH1 domain	6.4e-05	29.8
143	zf-DHHC	DHHC zinc finger domain	0.033	−11.5
147	zf-C2H2	Zinc finger, C2H2 type	9.2e-82	285.0
149	7tm_2	7 transmembrane receptor (Secretin family)	1.2e-22	88.7
151	lectin_c	Lectin C-type domain	0.0097	3.8
152	PDZ	PDZ domain (Also known as DHR or GLGF).	0.0031	24.2
153	cadherin	Cadherin domain	3.7e-95	329.6
155	COesterase	Carboxylesterases	7e-48	166.8
156	DSPc	Dual specificity phosphatase, catalytic domain	2.8e-29	110.7
157	efhand	EF hand	2.2e-14	61.2
159	A2M	Alpha-2-macroglobulin family	2.2e-07	25.5
160	zf-C2H2	Zinc finger, C2H2 type	8.8e-68	238.6
161	Nucleoside_tra2	Na+ dependent nucleoside transporter	8e-188	637.4
166	ras	Ras family	5.5e-31	116.4
167	zf-C3HC4	Zinc finger, C3HC4 type (RING finger)	3.2e-06	24.4
168	LRR	Leucine Rich Repeat	1.3e-06	35.4
169	RasGAP	GTPase-activator protein for Ras-like GTPase	6.4e-28	106.2
172	LMWPc	Low molecular weight phosphotyrosine protein phosphatase	9.7e-56	198.6
175	lectin_c	Lectin C-type domain	5.4e-06	33.3
178	PK	Pyruvate kinase	5.4e-12	46.3
179	vwa	von Willebrand factor type A domain	4.1e-63	223.1
180	neur_chan	Neurotransmitter-gated ion-channel	1.5e-120	413.9
181	C4	C-terminal tandem repeated domain in type 4 procollagen	1.4e-148	507.0
187	zf-C2H2	Zinc finger, C2H2 type	1.2e-25	98.6
188	Ribosomal_S2	Ribosomal protein S2	3.3e-79	276.5
191	ATP-synt_ab	ATP synthase alpha/beta family	5.7e-09	−1.3
192	ATP-synt_ab	ATP synthase alpha/beta family	3.1e-11	49.7
193	ank	Ank repeat	3.5e-33	123.7
195	GTP_EFTU	Elongation factor Tu family	8.1e-33	113.3
197	RF-1	Peptidyl-tRNA hydrolase domain	0.00034	10.7
200	WD40	WD domain, G-beta repeat	1e-05	32.4
201	Band_41	FERM domain (Band 4.1 family)	5.3e-86	269.7
205	pkinase	Eukaryotic protein kinase domain	1.6e-90	314.1
207	SCP	SCP-like extracellular protein	1e-15	60.4
208	ras	Ras family	1.5e-15	40.0
211	6PF2K	6-phosphofructo-2-kinase	5.2e-152	518.4
212	zf-C2H2	Zinc finger, C2H2 type	3e-104	359.8
215	pkinase	Eukaryotic protein kinase domain	3.2e-92	319.8
216	Oxysterol_BP	Oxysterol-binding protein	4.3e-48	173.2
219	pentaxin	Pentaxin family	2.6e-40	142.7
220	zf-C3HC4	Zinc finger, C3HC4 type (RING finger)	6.2e-08	30.0
221	Peptidase_M1	Peptidase family M1	1.5e-182	529.4
222	ig	Immunoglobulin domain	3.1e-07	28.4
224	BTB	BTB/POZ domain	1.8e-27	104.7
225	F-box	F-box domain.	3.5e-05	30.6
229	Na_Ca_Ex	Sodium/calcium exchanger protein	1.1e-23	92.1
231	lactamase_B	Metallo-beta-lactamase superfamily	0.01	−5.3
232	fibrinogen_C	Fibrinogen beta and gamma chains, C-terminal globular domain	7.5e-40	140.2
233	MIF	Macrophage migration inhibitory factor (MIF)	5.4e-66	232.7
234	SSF	Sodium:solute symporter family	1.7e-234	792.4
235	Cation_efflux	Cation efflux family	2.2e-63	224.0
237	AAA	ATPases associated with various cellular activities (AAA)	2.6e-85	296.8
238	HCO3_cotransp	HCO3- transporter family	0	1395.3
239	homeobox	Homeobox domain	1.1e-14	62.2
243	GST	Glutathione S-transferases.	0.0024	14.4
244	Viral_helicasel	Viral (Superfamily 1) RNA helicase	0.0019	14.7
245	Na_Galacto_symp	Sodium:galactoside symporter family	0.0068	−94.4
246	PPR	PPR repeat	0.0024	24.5
249	ENV_polyprotein	ENV polyprotein (coat polyprotein)	3.9e-45	155.2
252	UQ_con	Ubiquitin-conjugating enzyme	7.6e-15	62.8
254	7tm_1	7 transmembrane receptor (rhodopsin family)	2e-32	105.2
255	7tm_1	7 transmembrane receptor (rhodopsin family)	9.6e-43	137.9
256	UBA	UBA domain	1.2e-08	42.1
259	transmembrane4	Transmembrane 4 family	4.2e-44	144.0
267	thyroglobulin_1	Thyroglobulin type-1 repeat	3.2e-35	130.4
268	annexin	Annexin	6.7e-80	278.9
269	annexin	Annexin	8.5e-122	418.0
270	annexin	Annexin	6.7e-80	278.9
271	Armadillo_seg	Armadillo/beta-catenin-like repeats	6.4e-06	33.1
273	p450	Cytochrome P450	2.3e-132	453.1
274	p450	Cytochrome P450	5.9e-52	186.0
275	A2M_N	Alpha-2-macroglobulin family N-terminal region	2.1e-72	247.4
277	Thymosin	Thymosin beta-4 family	2.3e-16	67.8
279	WD40	WD domain, G-beta repeat	5.8e-19	76.4
288	F-box	F-box domain.	0.012	22.3
289	LRR	Leucine Rich Repeat	1.3e-17	72.0
290	TPR	TPR Domain	7.5e-54	192.3
293	cadherin	Cadherin domain	8.2e-07	36.1
297	UCH-1	Ubiquitin carboxyl-terminal hydrolases family 2	4.9e-08	40.1
298	ank	Ank repeat	9.1e-74	258.5
302	Clq	Clq domain	0.023	14.1
304	laminin_Nterm	Laminin N-terminal (Domain VI)	3.2e-52	186.9
305	enolase	Enol-ase	2.4e-69	243.8
307	Nol1_Nop2_Sun	NOL1/NOP2/sun family	0.0015	15.2
308	HIT	HIT family	3.2e-50	180.3
309	ank	Ank repeat	1e-20	82.3
310	lectin_c	Lectin C-type domain	8.8e-19	75.8
311	gpdh	glyceraldehyde 3-phosphate dehydrogenases	8.4e-237	793.9
314	mito_carr	Mitochondrial carrier proteins	2e-59	206.6
315	serpin	Serpins (serine protease inhibitors)	1.5e-109	372.9
316	MHC_I	Class I Histocompatibility antigen, domains alpha 1 and 2	5.1e-141	481.9
317	MHC_I	Class I Histocompatibility antigen, domains alpha 1 and 2	1.2e-132	454.1
318	MHC_I	Class I Histocompatibility antigen, domains alpha 1 and 2	3.2e-122	419.4
319	MHC_I	Class I Histocompatibility antigen, domains alpha 1 and 2	1.3e-119	410.8
320	MHC_I	Class I Histocompatibility antigen, domains alpha 1 and 2	1.8e-122	420.3
321	MHC_I	Class I Histocompatibility antigen, domains alpha 1 and 2	1.7e-131	450.2
323	MHC_I	Class I Histocompatibility antigen, domains alpha 1 and 2	4e-144	492.2
324	MHC_I	Class I Histocompatibility antigen, domains alpha 1 and 2	3.8e-105	362.7
325	MHC_I	Class I Histocompatibility antigen, domains alpha 1 and 2	1.3e-139	477.2
326	Kelch	Kelch motif	1.4e-101	350.9
328	Glycos_transf_2	Glycosyl transferases	1.9e-09	44.8
332	ig	Immunoglobulin domain	3.3e-07	28.3
333	UCH-1	Ubiquitin carboxyl-terminal hydrolases family 2	1.1e-12	55.6
336	ATP-gua_Ptrans	ATP:guanido phosphotransferase	4.1e-15	56.6
337	UBX	UBX domain	6.5e-18	72.9
338	UBX	UBX domain	6.5e-18	72.9
339	DnaJ	DnaJ domain	1e-36	135.4
340	zf-C3HC4	Zinc finger, C3HC4 type (RING finger)	5.3e-07	26.9
341	PKD	PKD domain	0	1485.5
342	CNH	CNH domain	2.7e-24	94.2
344	RhoGAP	RhOGAP domain	1.8e-59	211.0
345	Peptidase_M10	Matrixin	8.2e-110	378.2
346	Peptidase_M10	Matrixin	8.2e-110	378.2
349	UQ_con	Ubiquitin-conjugating enzyme	2.6e-06	0.9
350	ig	Immunoglobulin domain	1.6e-236	767.1
351	CNG_membrane	Transmembrane region cyclic Nucleotide Gated Channel	3.7e-108	372.7
353	RGS	Regulator of G protein signaling domain	4.6e-49	176.4
354	THF_DHG_CYH	Tetrahydrofolate dehydrogenase/cyclohydrolase	5e-106	365.7
355	gln-synt	Glutamine synthetase	1.9e-194	612.3
356	gln-synt	Glutamine synthetase	2e-39	125.5
357	SCAN	SCAN domain	9.7e-61	215.2
358	UPF0117	Domain of unknown function DUF36	1.3e-38	131.0
359	Trans_recep	Transient receptor	0	1115.3
362	Ammonium_transp	Ammonium Transporter Family	1.9e-56	200.9
363	Kelch	Kelch motif	3.2e-50	180.3
365	zf-C3HC4	Zinc finger, C3HC4 type (RING finger)	7.4e-26	88.2
366	SRCR	Scavenger receptor cysteine-rich domain	1.7e-25	98.1
367	adn_short	short chain dehydrogenase	1.1e-37	138.6
369	ANP	Atrial natriuretic peptide	1.5e-51	183.0
370	EGF	EGF-like domain	3.6e-26	100.4
372	STphosphatase	Ser/Thr protein phosphatase	1e-31	112.7
373	FKBP	FKBP-type peptidyl-prolyl cis-trans isomerases	1.7e-57	185.6
374	PX	PX domain	7.6e-16	66.1
375	ras	Ras family	5.2e-16	45.4
376	pkinase	Eukaryotic protein kinase domain	1.9e-56	200.9
377	pkinase	Eukaryotic protein kinase domain	1.9e-56	200.9
379	Peptidase_C1	Papain family cysteine protease	4.6e-119	409.0
380	Peptidase_C1	Papain family cysteine protease	7.4e-109	375.1
382	ig	Immunoglobulin domain	8.7e-10	36.6
384	Sec7	Sec7 domain	4.5e-71	249.5
385	cadherin	Cadherin domain	2.7e-95	330.0
387	lipocalin	Lipocalin/cytosolic fatty-acid binding protein family	1e-35	127.1
388	SH3	SH3 domain	1.5e-10	48.4
390	UCH-2	Ubiquitin carboxyl-terminal hydrolase family 2	1.8e-20	81.4
392	RCC1	Regulator of chromosome condensation (RCC1)	1.5e-14	56.1
397	crystall	Beta/Gamma crystallin	3.3e-38	140.4
403	PHD	PHD-finger	6.7e-15	62.9
404	ferritin	Ferritins	4.1e-114	386.1
405	CUB	CUB domain	2.4e-13	57.8
406	ATP-gua_Ptrans	ATP:guanido phosphotransferase	5e-05	20.7
407	SPRY	SPRY domain	2e-09	44.8
410	ELM2	ELM2 domain	9.4e-15	62.4
411	Reprolysin	Reprolysin (M12B) family zinc metalloprotease	7.2e-15	56.4
415	ank	Ank repeat	3.5e-18	73.8
418	C2	C2 domain	1e-75	264.9
419	C2	C2 domain	1e-75	264.9
420	p450	Cytochrome P450	9.8e-120	411.2
422	aa_permeases	Amino acid permease	1.3e-08	−108.1
424	kinesin	Kinesin motor domain	1.9e-115	397.0
425	LRR	Leucine Rich Repeat	8.3e-26	99.2
426	kinesin	Kinesin motor domain	8e-63	222.1
427	zf-DHHC	DHHC zinc finger domain	9.1e-34	125.6
428	EGF	EGF-like domain	3.2e-45	163.6
432	TWIK_channel	TASK K+ channel	1.8e-09	7.9
433	pkinase	Eukaryotic protein kinase domain	2.3e-78	273.8
434	PH	PH domain	0.00018	21.0
436	ig	Immunoglobulin domain	9.2e-09	33.3
440	LRR	Leucine Rich Repeat	2.8e-16	67.5
441	MACPF	MAC/Perforin domain	0.016	−71.3
444	efhand	EF hand	0.00027	27.7
448	EPH_lbd	Ephrin receptor ligand binding domain	7.9e-135	461.3
456	C1q	C1q domain	0.023	14.1
458	ASC	Amiloride-sensitive sodium channel	9.2e-127	434.5
459	pkinase	Eukaryotic protein kinase domain	0.083	10.4
460	PBP	Phosphatidylethanolamine-binding protein	1.2e-71	251.4
464	rrm	RNA recognition motif. (a.k.a. RRM, RBD, or RNP domain)	2.8e-12	54.2
466	DEP	Domain found in Dishevelled, Eg1-10, and Pleckstrin	6.7e-18	72.9
470	SCAN	SCAN domain	4.3e-52	186.5
471	EGF	EGF-like domain	5.1e-28	106.5
473	Ferric_reduct	Ferric reductase like transmembrane component	6.8e-74	258.9
474	zf-C2H2	Zinc finger, C2H2 type	5.7e-12	53.2
478	CAP_GLY	CAP-Gly domain	3.4e-46	166.9
477	PAP2	PAP2 superfamily	4.9e-10	46.8
476	SCAN	SCAN domain	9.3e-70	245.2
480	LRR	Leucine Rich Repeat	1e-13	59.0
483	PH	PH domain	1e-15	61.2
484	PH	PH domain	2.3e-21	81.3
489	RhoGAP	RhoGAP domain	5.7e-57	202.7
493	annexin	Annexin	4.7e-70	246.2
495	zf-C3HC4	Zinc finger, C3HC4 type (RING finger)	4.2e-06	24.0
497	TK	Thymidine kinases	9.4e-118	338.0
501	PHD	PHD-finger	0.01	8.3
504	rrm	RNA recognition motif. (a.k.a. RRM, RBD, or RNP domain)	1.3e-19	78.6
506	rvt	Reverse transcriptase (RNA-dependent DNA polymerase)	3.2e-30	113.8
508	Insulin	Insulin/IGF/Relaxin family	6.6e-22	86.2
510	COX5A	Cytochrome c oxidase subunit Va	1.2e-55	198.3
511	Guanylate_kin	Guanylate kinase	6.2e-38	139.4
513	hexokinase	Hexokinase	0	1029.0
516	trypsin	Trypsin	1.4e-78	250.0
519	Glycos_transf_1	Glycosyl transferases group 1	2.2e-27	102.8
520	EMP24_GP25L	emp24/gp25L/p24 family	3.5e-70	246.6
521	EMP24_GP25L	emp24/gp25L/p24 family	3.5e-81	283.1
522	14-3-3	14-3-3 proteins	6.6e-150	511.4
526	zf-CCCH	Zinc finger C-x8-C-x5-C-x3-H type (and similar).	0.039	13.4
527	SpoU_methylase	SpoU rRNA Methylase family	2.1e-27	104.5
529	MMR_HSR1	GTPase of unknown function	1.5e-90	314.2
531	RNase PH	3′ exoribonuclease family	2.2e-96	333.6
534	COLFI	Fibrillar collagen C-terminal domain	5.6e-50	129.2
535	COLFI	Fibrillar collagen C-terminal domain	3.1e-58	150.3
536	CH	Calponin homology (CH) domain	5.6e-14	59.9
541	FH2	Formin Homology 2 Domain	4.5e-07	−23.3
542	AAA	ATPases associated with various cellular activities (AAA)	5.5e-33	123.0
547	lectin_c	Lectin C-type domain	2.3e-29	111.0
551	DEAD	DEAD/DEAH box helicase	9.2e-58	185.6
554	C2	C2 domain	1.3e-51	184.9
555	ank	Ank repeat	1.5e-26	101.6
556	ank	Ank repeat	6e-137	468.4
559	zf-C3HC4	Zinc finger, C3HC4 type (RING finger)	1.1e-10	38.9
560	ig	Immunoglobulin domain	1.2e-81	268.2
562	COesterase	Carboxylesterases	1.4e-15	55.8
564	FYVE	FYVE zinc finger	3.8e-15	58.7
572	SPRY	SPRY domain	0.0059	2.8
573	LRR	Leucine Rich Repeat	0.0076	22.9
575	Skp1	Skp1 family	6.3e-10	46.4
576	RhoGAP	RhoGAP domain	4.2e-31	116.8
577	UQ_con	Ubiquitin-conjugating enzyme	6.3e-50	179.3
579	LRR	Leucine Rich Repeat	8.7e-34	125.7
580	K_tetra	K+ channel tetramerisation domain	0.0016	−5.0
581	RCC1	Regulator of chromosome condensation (RCC1)	8.1e-10	39.5
582	profilin	Profilins	5.4e-63	222.7
583	profilin	Profilins	4e-48	173.3
585	PHD	PHD-finger	0.041	2.8
586	kinesin	Kinesin motor domain	5.2e-114	392.2
588	aminotran_3	Aminotransferases class-III pyridoxal-phosphate	4e-75	217.6
593	TPR	TPR Domain	0.00036	27.3
594	PCMT	Protein-L-isoaspartate (D-aspartate) O-methyltransferase (PCMT)	1.6e-11	21.8
597	homeobox	Homeobox domain	6.1e-27	102.9
598	zf-C2H2	Zinc finger, C2H2 type	6.8e-85	295.5
599	Ribosomal_S5	Ribosomal protein S5	9.9e-12	45.7
600	IQ	IQ calmodulin-binding motif	1.8e-18	74.8
604	DUF6	Integral membrane protein DUF6	0.083	9.1
605	zf-C2H2	Zinc finger, C2H2 type	2.1e-05	31.4
606	F-box	F-box domain.	1.7e-05	31.7
607	Ran_BP1	RanBP1 domain.	1.1e-88	308.0
608	actin	Actin	4.4e-156	513.5
611	GTP_EFTU	Elongation factor Tu family	8.5e-22	76.1
619	carb_anhydrase	Eukaryotic-type carbonic anhydrase	2.5e-171	582.6
620	peroxidase	Peroxidase	5.8e-208	704.3
621	FAD_Gly3P_dh	FAD-dependent glycerol-3-phosphate dehydrogenase	0.029	−256.0
623	CH	Calponin homology (CH) domain	1.4e-25	98.4
625	mutT	Bacterial	4.3e-09	36.9
626	HECT	HECT-domain (ubiquitin-transferase).	4.3e-16	66.9
627	ig	Immunoglobulin domain	0.024	12.6
631	Acetyltransf	Acetyltransferase (GNAT) family	2.9e-12	54.2
632	Acetyltransf	Acetyltransferase (GNAT) family	2.9e-12	54.2
634	pyr_redox	Pyridine nucleotide-disulphide oxidoreductase	2.9e-05	20.5
639	Sm	Sm protein	1.3e-07	38.7
640	PDZ	PDZ domain (Also known as DHR or GLGF).	8.1e-15	62.7
643	Syntaxin	Syntaxin	2.3e-09	38.1
645	DnaJ	DnaJ domain	1.3e-39	145.1
646	HMG_box	HMG (high mobility group) box	2.7e-27	104.1
650	HMG_box	HMG (high mobiiity group) box	3.4e-30	113.7
653	DUF6	Integral membrane protein DUF6	3e-12	54.1
654	FF	FF domain	1.2e-33	125.2
655	FF	FF domain	1.2e-33	125.2
658	aminotran_5	Aminotransferases class-V	1.3e-127	437.3
659	aminotran_5	Aminotransferases class-V	3.5e-94	326.3
666	SAM_PNT	Sterile alpha motif (SAM)/Pointed domain	0.0021	6.9
667	DUF52	Protein of unknown function DUF52	4.7e-64	226.2
668	C2	C2 domain	6.8e-36	132.7
669	C2	C2 domain	7.8e-35	129.2
671	7tm_1	7 transmembrane receptor (rhodopsin family)	2.4e-50	162.1
672	K_tetra	K+ channel tetramerisation domain	1.1e-25	98.7
675	cyclin	Cyclin	7.1e-14	52.2
676	zf-C2H2	Zinc finger, C2H2 type	1.4e-18	75.2
679	PDZ	PDZ domain (Also known as DHR or GLGF).	4.3e-16	66.9
681	rrm	RNA recognition motif. (a.k.a. RRM, RBD, or RNP domain)	1.7e-47	171.2
689	Gelsolin	Gelsolin repeat.	8.5e-89	308.4
691	TBC	TBC domain	2e-08	15.3
692	TBC	TBC domain	2e-07	1.3
697	GTP_CDC	Cell division protein	2.3e-14	54.4
698	transmembrane4	Transmembrane 4 family	1e-38	126.8
705	Colipase	Colipase	4.9e-21	83.3
706	SH2	Src homology domain 2	1.9e-05	19.4
710	SUI1	Translation initiation factor SUI1	5e-48	173.0
715	SH3	SH3 domain	0.013	8.0
716	UBA	UBA domain	1.3e-09	45.4
717	Ribosomal_S21	Ribosomal protein S21	0.0039	11.7
718	transmembrane4	Transmembrane 4 family	3.6e-52	169.8
723	LRR	Leucine Rich Repeat	1.1e-48	175.2
724	UCH-2	Ubiquitin carboxyl-terminal hydrolase family 2	5.6e-28	106.4
725	F-box	F-box domain.	0.0016	25.1
727	FGGY	FGGY family of carbohydrate kinases	5.8e-62	219.3
732	pyr_redox	Pyridine nucleotide-disulphide oxidoreductase	4.2e-23	80.9
733	WD40	WD domain, G-beta repeat	0.073	19.6
735	Ribosomal_L44	Ribosomal protein L44	1e-38	142.1
738	WD40	WD domain, G-beta repeat	1.3e-08	42.1
742	dCMP_cyt_deam	Cytidine and deoxycytidylate deaminase zinc-binding region	1.7e-09	45.0
743	gntR	Bacterial regulatory proteins, gntR family	0.062	11.4
744	cpn60_TCP1	TCP-1/cpn60 chaperonin family	2.9e-74	260.1
748	zf-C2H2	Zinc finger, C2H2 type	6.5e-15	63.0
751	VHS	VHS domain	1.7e-61	217.7
752	rrm	RNA recognition motif. (a.k.a. RRM, RBD, or RNP domain)	4.2e-23	90.2
753	ank	Ank repeat	1.4e-80	281.1
757	cyclin	Cyclin	0.026	11.5
761	SH2	Src homology domain 2	5.6e-05	18.0
762	abhydrolase	alpha/beta hydrolase fold	1.2e-21	85.3
763	zf-C3HC4	Zinc finger, C3HC4 type (RING finger)	1.8e-05	22.0
764	WD40	WD domain, G-beta repeat	1e-05	32.4
766	Acyltransferase	Acyltransferase	0.00021	15.9
771	helicase_C	Helicases conserved C-terminal domain	3e-15	64.1
772	Na_Ca_Ex	Sodium/calcium exchanger protein	8e-76	265.3
773	vwa	von Willebrand factor type A domain	9.6e-29	108.9
775	DEAD	DEAD/DEAH box helicase	0.042	9.5
776	pkinase	Eukaryotic protein kinase domain	1.6e-07	31.0
780	rrm	RNA recognition motif. (a.k.a. RRM, RBD, or RNP domain)	1.1e-05	32.3
782	PX	PX domain	1.5e-23	91.7
783	PMP22_Claudin	PMP-22/EMP/MP20/Claudin family	1.5e-56	201.3
785	homeobox	Homeobox domain	0.00021	23.9
786	homeobox	Homeobox domain	0.00021	23.9
790	efhand	EF hand	3.6e-18	73.8
793	Glyco_hydro_31	Glycosyl hydrolases family 31	9.8e-18	62.0
797	Smr	Smr domain	0.0029	13.0
798	Smr	Smr domain	0.0029	13.0
799	mito_carr	Mitochondrial carrier proteins	3.1e-61	212.8
801	Oxysterol_BP	Oxysterol-binding protein	3.4e-78	273.2
803	lipoxygenase	Lipoxygenase	1.7e-140	480.2
805	Rho_GDI	RHO protein GDP dissociation inhibitor	1.1e-122	420.9
809	homeobox	Homeobox domain	3.9e-32	120.2
810	lipocalin	Lipocalin/cytosolic fatty-acid binding protein family	8.5e-31	110.2
811	ig	Immunoglobulin domain	8.8e-14	49.5
814	Keratin_B2	Keratin, high sulfur B2 protein	3.3e-07	32.9
815	sugar_tr	Sugar (and other) transporter	0.0057	−109.4
817	VPS9	Vacuolar sorting protein 9 (VPS9) domain	1.1e-37	138.7
818	SSF	Sodium:solute symporter family	5.9e-206	697.6
819	Peptidase_M3	Peptidase family M3	1.4e-280	945.5
820	RasGAP	GTPase-activator protein for Ras-like GRPase	1.3e-26	101.9
824	zf-C2H2	Zinc finger, C2H2 type	2e-87	303.9
825	ATP-synt_C	ATP synthase subunit C	5.4e-08	40.0
826	zf-C3HC4	Zinc finger, C3HC4 type (RING finger)	4.7e-13	46.6
827	FH2	Formin Homology 2 Domain	9e-55	195.4
833	SH3	SH3 domain	9.2e-14	59.2
835	SET	SET domain	9.9e-52	185.3
836	zf-DHHC	DHHC zinc finger domain	2.4e-25	97.7
837	zf-C2H2	ZinC finger, C2H2 type	8.2e-14	59.3
840	Trans_recep	Transient receptor	0	1115.3
850	Ribosomal_S12	Ribosomal protein S12	1e-24	93.8
851	Acyltransferase	Acyltransferase	7.3e-06	32.2
853	zf-C2H2	Zinc finger, C2H2 type	5.1e-80	279.2
854	lectin_c	Lectin C-type domain	2.8e-24	94.1
855	lectin_c	Lectin C-type domain	2.8e-24	94.1
856	efhand	EF hand	9.7e-10	45.8
858	LRR	Leucine Rich Repeat	3.1e-09	44.1
868	filament	Intermediate filament proteins	0.00027	20.0
869	MAGE	MAGE family	2.3e-19	77.8
871	Hydrolase	haloacid dehalogenase-like hydrolase	0.00056	12.6
872	Fe_hyd_SSU	Iron hydrogenase small subunit	0.00025	21.9
873	filament	Intermediate filament proteins	3.7e-143	489.0
874	Keratin_B2	Keratin, high sulfur B2 protein	0.082	−51.3
876	metalthio	Metallothionein	4.7e-23	90.0
877	C2	C2 domain	4.7e-53	189.7
880	efhand	EF hand	0.041	20.5
885	EGF	EGF-like domain	1.1e-32	122.0
886	7tm_1	7 transmembrane receptor (rhodopsin family)	0.003	12.5
887	rrm	RNA recognition motif. (a.k.a. RRM, RBD, or RNP domain)	0.058	15.6
888	Kelch	Kelch motif	5.3e-22	86.5
889	tsp_1	Thrombospondin type 1 domain	0.0014	23.3
890	adh_zinc	Zinc-binding dehydrogenases	1.5e-55	197.9
891	pkinase	Eukaryotic protein kinase domain	3e-05	22.8
893	zf-C2H2	Zinc finger, C2H2 type	2.1e-30	114.5
895	V_ATPase_sub_a	V-type ATPase 116kDa subunit family	0	1263.1
896	aa_permeases	Amino acid permease	5.6e-08	−125.3
900	MCT	Monocarboxylate transporter	1e-42	155.3
901	Filamin	Filamin/ABP280 repeat.	2.3e-20	81.1
905	ank	Ank repeat	3.3e-88	306.5
906	ras	Ras family	1.4e-13	17.8
907	Acyltransferase	Acyltransferase	6.4e-34	126.1
908	wap	WAP-type (Whey Acidic Protein) ‘four-disulfide core’	0.039	7.8
909	arf	ADP-ribosylation factor family	3.7e-11	25.6
910	zf-C2H2	Zinc finger, C2H2 type	2.5e-09	44.4
911	zf-C2H2	Zinc finger, C2H2 type	5.6e-20	79.8
912	BTB	BTB/POZ domain	1.1e-27	105.4
913	SNF2_N	SNF2 and others N-terminal domain	1.7e-71	250.9
914	ras	Ras family	2.4e-69	243.8
916	rrm	RNA recognition motif. (a.k.a. RRM, RBD, or RNP domain)	0.061	15.4
919	T-box	T-box	2.7e-45	159.8
923	PH	PH domain	1.5e-14	57.0
924	isodh	Isocitrate and isopropylmalate dehydrogenases	3.7e-125	421.5
925	ank	Ank repeat	2.5e-08	41.1
926	ank	Ank repeat	2.5e-08	41.1
928	DUF51	Protein of unknown function DUF51	5.7e-13	52.5
929	zf-C2H2	Zinc finger, C2H2 type	2.1e-72	253.9
930	AAA	ATPases associated with various cellular activities (AAA)	1.1e-67	238.3
931	SSF	Sodium:solute symporter family	4.8e-170	578.3
932	SSF	Sodium:solute symporter family	1.3e-198	673.2
934	aminotran_1	Aminotransferases class-I	2e-10	10.6
935	DSPc	Dual specificity phosphatase, catalytic domain	1.1e-29	112.1
936	ig	Immunoglobulin domain	2.7e-15	54.3
937	MCM	MCM2/3/5 family	1.9e-11	−67.4
938	Kelch	Kelch motif	5.7e-56	199.4
939	UBA	UBA domain	2.7e-06	34.3
940	UBA	UBA domain	2.7e-06	34.3
941	UBA	UBA domain	2.7e-06	34.3
942	Na_H_Exchanger	Sodium/hydrogen exchanger family	0.00067	−108.1
943	Acyl-CoA_hydro	Cytosolic long-chain acyl-CoA thioester hydrolase	9e-74	258.5
944	zf-DHHC	DHHC zinc finger domain	8.6e-18	72.5
945	filament	Intermediate filament proteins	5.6e-29	109.7
946	SPRY	SPRY domain	0.022	−3.0
947	dsrm	Double-stranded RNA binding motif	1.5e-12	55.1
948	SPRY	SPRY domain	2.8e-11	50.9

TABLE 5
	POSITION
	OF SIGNAL	maxS
	IN AMINO ACID	(MAXIMUM	meanS
SEQ ID NO:	SEQUENCE	SCORE)	(MEAN SCORE)
1	1-24	0.926	0.738
8	1-48	0.994	0.655
9	1-31	0.984	0.921
11	1-36	0.994	0.757
12	1-20	0.977	0.902
16	1-25	0.921	0.787
17	1-33	0.967	0.803
22	1-26	0.949	0.664
24	1-28	0.929	0.700
26	1-17	0.919	0.828
31	1-19	0.890	0.552
33	1-19	0.981	0.916
35	1-21	0.980	0.904
36	1-21	0.980	0.904
38	1-26	0.951	0.801
41	1-43	0.994	0.659
44	1-19	0.942	0.693
52	1-21	0.989	0.925
57	1-18	0.964	0.812
58	1-19	0.972	0.915
59	1-24	0.997	0.929
60	1-16	0.945	0.737
61	1-31	0.957	0.775
68	1-28	0.988	0.938
69	1-23	0.976	0.897
72	1-33	0.948	0.776
75	1-31	0.991	0.925
76	1-41	0.942	0.703
77	1-36	0.910	0.749
79	1-27	0.962	0.696
82	1-24	0.943	0.832
86	1-27	0.962	0.856
87	1-19	0.967	0.909
88	1-39	0.986	0.922
89	1-28	0.982	0.924
91	1-29	0.984	0.763
92	1-22	0.974	0.796
93	1-29	0.928	0.725
94	1-44	0.995	0.811
95	1-36	0.901	0.766
97	1-25	0.921	0.787
103	1-23	0.966	0.812
106	1-19	0.951	0.895
107	1-16	0.927	0.827
108	1-25	0.949	0.823
110	1-28	0.980	0.848
113	1-24	0.965	0.891
114	1-25	0.946	0.860
119	1-36	0.964	0.648
126	1-32	0.941	0.669
128	1-17	0.995	0.974
135	1-18	0.968	0.799
141	1-24	0.882	0.599
143	1-38	0.991	0.904
146	1-29	0.963	0.888
148	1-19	0.892	0.715
153	1-34	0.921	0.652
154	1-20	0.951	0.839
158	1-31	0.921	0.659
162	1-36	0.992	0.917
176	1-30	0.989	0.910
177	1-28	0.974	0.851
179	1-25	0.937	0.812
182	1-30	0.978	0.786
183	1-27	0.987	0.879
185	1-23	0.923	0.655
196	1-28	0.980	0.893
199	1-27	0.963	0.833
202	1-24	0.976	0.913
203	1-24	0.988	0.967
204	1-22	0.968	0.831
206	1-21	0.952	0.822
207	1-42	0.939	0.682
209	1-22	0.984	0.928
210	1-22	0.984	0.928
217	1-21	0.942	0.713
219	1-18	0.922	0.838
222	1-18	0.988	0.944
226	1-18	0.975	0.958
227	1-18	0.975	0.958
228	1-18	0.975	0.958
229	1-48	0.989	0.889
230	1-23	0.996	0.936
232	1-16	0.967	0.933
245	1-15	0.948	0.907
247	1-27	0.936	0.689
248	1-42	0.978	0.750
249	1-15	0.977	0.966
251	1-26	0.976	0.875
252	1-28	0.973	0.822
253	1-28	0.990	0.925
257	1-22	0.982	0.933
258	1-15	0.986	0.919
259	1-27	0.994	0.900
267	1-21	0.989	0.871
272	1-28	0.976	0.653
278	1-20	0.987	0.916
281	1-30	0.996	0.894
282	1-41	0.983	0.791
286	1-20	0.978	0.893
291	1-17	0.953	0.784
292	1-25	0.950	0.897
293	1-20	0.974	0.912
294	1-15	0.974	0.817
299	1-35	0.973	0.795
302	1-22	0.982	0.872
303	1-18	0.983	0.927
306	1-20	0.934	0.828
307	1-16	0.952	0.807
308	1-19	0.904	0.656
312	1-35	0.957	0.640
313	1-35	0.957	0.640
315	1-33	0.953	0.707
316	1-24	0.981	0.884
317	1-24	0.987	0.914
318	1-21	0.977	0.905
319	1-24	0.978	0.911
320	1-18	0.984	0.958
321	1-18	0.984	0.958
322	1-24	0.989	0.922
323	1-18	0.984	0.956
324	1-18	0.986	0.965
325	1-18	0.986	0.965
326	1-32	0.956	0.706
329	1-48	0.983	0.616
330	1-20	0.965	0.878
334	1-16	0.921	0.828
335	1-20	0.937	0.700
345	1-19	0.995	0.971
346	1-19	0.995	0.971
348	1-20	0.926	0.751
354	1-29	0.981	0.937
362	1-27	0.977	0.849
366	1-24	0.977	0.845
367	1-23	0.990	0.833
369	1-25	0.971	0.894
370	1-16	0.961	0.916
371	1-41	0.980	0.681
379	1-17	0.977	0.921
380	1-17	0.977	0.921
381	1-26	0.993	0.894
383	1-25	0.986	0.939
385	1-33	0.977	0.811
392	1-43	0.992	0.943
393	1-20	0.943	0.882
395	1-20	0.995	0.933
396	1-26	0.938	0.663
398	1-21	0.955	0.767
399	1-19	0.920	0.692
400	1-41	0.937	0.604
401	1-41	0.937	0.604
405	1-19	0.986	0.961
409	1-41	0.923	0.559
411	1-25	0.973	0.853
413	1-20	0.935	0.817
416	1-29	0.958	0.637
417	1-23	0.991	0.740
420	1-29	0.986	0.848
421	1-18	0.997	0.979
425	1-16	0.979	0.964
427	1-25	0.980	0.952
428	1-37	0.989	0.822
430	1-37	0.984	0.878
431	1-42	0.978	0.698
432	1-18	0.969	0.913
435	1-31	0.981	0.856
438	1-36	0.987	0.595
439	1-27	0.955	0.786
441	1-17	0.915	0.825
442	1-34	0.954	0.783
445	1-19	0.941	0.839
448	1-27	0.944	0.778
449	1-20	0.941	0.734
456	1-22	0.982	0.872
479	1-19	0.993	0.931
480	1-22	0.992	0.807
482	1-22	0.918	0.716
485	1-38	0.994	0.887
488	1-24	0.914	0.588
490	1-28	0.990	0.919
494	1-26	0.990	0.969
498	1-36	0.954	0.817
502	1-23	0.896	0.747
503	1-23	0.969	0.855
508	1-24	0.985	0.932
514	1-23	0.985	0.951
516	1-27	0.985	0.927
517	1-21	0.960	0.649
519	1-41	0.990	0.922
520	1-20	0.991	0.954
521	1-20	0.991	0.954
525	1-21	0.975	0.909
530	1-11	0.900	0.758
545	1-18	0.933	0.634
547	1-22	0.899	0.639
548	1-40	0.953	0.668
561	1-16	0.881	0.607
568	1-36	0.924	0.590
569	1-25	0.919	0.718
570	1-28	0.928	0.590
571	1-27	0.967	0.872
578	1-25	0.998	0.934
579	1-30	0.973	0.829
599	1-20	0.937	0.728
603	1-30	0.962	0.801
604	1-17	0.925	0.779
609	1-20	0.981	0.910
614	1-17	0.977	0.921
617	1-25	0.938	0.677
620	1-22	0.975	0.822
629	1-34	0.934	0.552
630	1-28	0.946	0.799
635	1-15	0.954	0.725
636	1-30	0.938	0.808
644	1-24	0.973	0.910
649	1-24	0.920	0.596
652	1-29	0.968	0.769
656	1-28	0.975	0.926
665	1-25	0.977	0.776
670	1-41	0.986	0.847
679	1-24	0.915	0.578
682	1-24	0.950	0.737
683	1-28	0.987	0.785
684	1-19	0.890	0.552
685	1-22	0.968	0.934
686	1-22	0.968	0.934
687	1-23	0.965	0.883
688	1-23	0.965	0.883
690	1-26	0.896	0.615
693	1-30	0.956	0.665
696	1-39	0.971	0.694
698	1-44	0.992	0.576
701	1-45	0.964	0.657
702	1-45	0.964	0.657
705	1-17	0.968	0.947
707	1-28	0.960	0.607
709	1-31	0.977	0.720
714	1-28	0.956	0.604
718	1-47	0.985	0.646
719	1-19	0.990	0.946
729	1-47	0.996	0.556
736	1-18	0.930	0.679
739	1-25	0.992	0.948
741	1-26	0.947	0.594
745	1-22	0.963	0.859
747	1-26	0.956	0.830
755	1-26	0.990	0.959
758	1-40	0.987	0.917
759	1-26	0.985	0.917
762	1-36	0.991	0.868
765	1-24	0.887	0.553
766	1-23	0.995	0.974
767	1-18	0.997	0.977
768	1-22	0.968	0.934
770	1-26	0.974	0.730
773	1-18	0.983	0.939
783	1-24	0.988	0.919
795	1-30	0.939	0.639
796	1-26	0.984	0.746
807	1-23	0.965	0.693
810	1-22	0.962	0.919
811	1-21	0.988	0.911
812	1-36	0.980	0.559
821	1-10	0.880	0.780
823	1-14	0.922	0.678
825	1-39	0.982	0.829
830	1-34	0.973	0.817
831	1-36	0.976	0.794
838	1-18	0.918	0.651
839	1-35	0.991	0.834
841	1-22	0.947	0.677
847	1-24	0.963	0.865
848	1-30	0.967	0.758
849	1-33	0.926	0.807
852	1-25	0.889	0.718
857	1-20	0.995	0.968
859	1-27	0.887	0.642
862	1-41	0.975	0.875
865	1-26	0.921	0.620
878	1-36	0.951	0.782
884	1-19	0.983	0.888
885	1-23	0.971	0.941
886	1-40	0.964	0.560
891	1-39	0.942	0.587
898	1-16	0.945	0.737
899	1-24	0.946	0.593
900	1-44	0.974	0.662
902	1-26	0.974	0.730
903	1-27	0.952	0.832
908	1-25	0.960	0.642
918	1-31	0.956	0.846
921	1-16	0.968	0.921
936	1-19	0.984	0.936
944	1-35	0.954	0.699

SEQUENCE LISTING
The patent contains a lengthy “Sequence Listing” section. A copy of the “Sequence Listing” is available in electronic form from the USPTO
web site (http://seqdata.uspto.gov/sequence.html?DocID=06783969B1). An electronic copy of the “Sequence Listing” will also be available from the
USPTO upon request and payment of the fee set forth in 37 CFR 1.19(b)(3).

Claims

1. An isolated polypeptide encoded by the polynucleotide of SEQ ID NO: 380.

2. A composition comprising the polypeptide of claim 1 and a carrier.

3. The polypeptide of claim 1 wherein the polypeptide is provided on a polypeptide array.