Claims
- 1. An isolated nucleic acid selected from the group consisting of:
(a) the DNA sequence of SEQ ID NO:1; (b) an isolated nucleic acid encoding an amino acid sequence comprising the sequence of SEQ ID NO:2; (c) an isolated nucleic acid that hybridizes to either strand of a denatured, DNA molecule comprising the nucleic acid sequence of (a) or (b) under conditions of moderate stringency; (d) an isolated nucleic acid derived by in vitro mutagenesis from SEQ ID NO:1; (e) an isolated nucleic acid degenerate from SEQ ID NO:1 as a result of the genetic code; and (f) an isolated nucleic acid selected from the group consisting of mouse DAKAR DNA, human DAKAR DNA, an allelic variant of mouse DAKAR DNA, an allelic variant of human DAKAR DNA, and a species homolog of DAKAR DNA.
- 2. A recombinant vector that directs the expression of the nucleic acid of claim 1.
- 3. An isolated polypeptide encoded by the nucleic acid of claim 1.
- 4. An isolated polypeptide according to claim 3 having a molecular weight of approximately 87 kD as determined by SDS-PAGE.
- 5. An isolated polypeptide according to claim 3 in non-glycosylated form.
- 6. Isolated antibodies that bind to a polypeptide of claim 3.
- 7. Isolated antibodies according to claim 6, wherein the antibodies are monoclonal antibodies.
- 8. A host cell transfected or transduced with the vector of claim 2.
- 9. A method for the production of a DAKAR polypeptide comprising culturing a host cell of claim 2 in suitable culture medium under conditions promoting gene expression, and recovering said polypeptide from the culture medium.
- 10. The method of claim 9, wherein the host cell is selected from the group consisting of bacterial cells, yeast cells, plant cells, and animal cells.
- 11. A method for determining the molecular weight of a sample protein comprising comparison of the molecular weight of a sample protein with the molecular weight of a polypeptide of claim 3, wherein said comparison of molecular weights comprises:
application of said sample protein and said polypeptide to an acrylamide gel; resolution of said sample protein and said polypeptide using an electric current; and application of a reagent for detecting said sample protein and said polypeptide.
- 12. A kit for the determining the molecular weights of peptide fragments of a sample protein comprising a vessel;
a polypeptide of claim 3;at least one enzyme selected from the group consisting of Asparaginylendopeptidase, Arginylendopeptidase, Achrombobacter protease I, Trypsin, Staphylococcus aureus V8 protease, Endoproteinase Asp-N, and Endoproteinase Lys-C; a mutant of said polypeptide derived by in vitro mutagenesis, wherein a site of enzymatic cleavage on said polypeptide of said enzyme has been removed; and fragmented peptides derived from said polypeptide by enzymatic cleavage with the selected enzyme wherein said polypeptide and said sample protein are contacted with said enzyme, and said protein, polypeptides, and fragmented peptides are detected by a process comprising the steps of application of said sample protein, polypeptides, and fragmented peptides to an acrylamide gel; resolution of the protein, polypeptides, and fragmented peptides using an electric current; and application of a reagent for the detection of said sample protein, polypeptides and fragmented peptides.
- 13. A DAKAR polypeptide comprising an amino acid sequence selected from the group consisting of:
(a) amino acids 1-785 of SEQ ID NO:2; (b) amino acids 17-297 of SEQ ID NO:2; and (c) amino acids 471-768 of SEQ ID NO:2.
- 14. An assay of DAKAR protein kinase activity where the kinase moiety is a purified polypeptide as described in claims 3, 4, 5, or 13.
- 15. The nucleic acid of claim 1 wherein said conditions of moderate stringency comprise hybridization in 50% formamide and 6×SSC at 42° C. and washing in 0.5×SSC and 0.1% SDS at 60° C.
- 16. The polypeptide of claim 3, wherein said polypeptide is a protein kinase.
- 17. A method of inhibiting the kinase activity of DAKAR in a mammal, said method comprising administering to said mammal an effective amount of a compound that inhibits the kinase activity of a polypeptide comprising the sequence of SEQ ID NO:2 or a fragment thereof.
- 18. A method of enhancing the kinase activity of DAKAR in a mammal, said method comprising administering to said mammal an effective amount of a compound that enhances the kinase activity of a polypeptide comprising the sequence of SEQ ID NO:2 or a fragment thereof.
- 19. A method of treating an animal having a disease characterized by an overproduction or upregulated production of DAKAR, said method comprising administering to said mammal a composition comprising an effective amount of a compound that inhibits the kinase activity of a polypeptide comprising the sequence of SEQ ID NO:2 or a fragment thereof.
- 20. A method of treating an animal having a disease characterized by an underproduction or downregulated production of DAKAR, said method comprising administering to said mammal a composition comprising an effective amount of a compound that enhances the kinase activity of a polypeptide comprising the sequence of SEQ ID NO:2 or a fragment thereof.
- 21. A method of designing an molecule that inhibits the kinase activity of a polypeptide according to claim 16, said method comprising the steps of:
determining the three dimensional structure of said polypeptide; analyzing said three dimensional structure for the likely binding sites of substrates; synthesizing a molecule that incorporates a predictive reactive site; and determining the kinase-inhibiting activity of the molecule.
- 22. A method of designing a molecule that increases the kinase activity of a polypeptide according to claim 16, said method comprising the steps of:
determining the three dimensional structure of said polypeptide; analyzing said three dimensional structure for the likely binding sites of substrates; synthesizing a molecule that incorporates a predictive reactive site; and determining the kinase-enhancing activity of the molecule.
CROSS-REFERENCE TO RELATED APPLICATION
[0001] The present invention is related to U.S. provisional application No. 60/119,353, filed Feb. 9, 1999, which is based on U.S. provisional application No. 60/099,973, filed Sep. 11, 1998, and on U.S. provisional application No. 60/095,269, filed Aug. 4, 1998. Each of these applications is incorporated herein by reference in its entirety.
Provisional Applications (3)
|
Number |
Date |
Country |
|
60095269 |
Aug 1998 |
US |
|
60099973 |
Sep 1998 |
US |
|
60119353 |
Feb 1999 |
US |
Divisions (1)
|
Number |
Date |
Country |
Parent |
09509802 |
Jun 2000 |
US |
Child |
10299327 |
Nov 2002 |
US |