Increased activity and efficiency of expansin-like proteins

BACKGROUND OF THE INVENTION

Cell wall proteins play important roles in regulating cell wall extensibility which in turn controls cell enlargement. Among cell wall proteins studied to date, expansins are unique in their ability to induce immediate cell wall extension in vitro and cell expansion in vivo. Expansins are extracellular proteins that promote plant cell wall enlargement, evidently by disrupting noncovalent bonding between cellulose microfibrils and matrix polymers (McQueen-Mason, S., et al., (1994) Proc. Natl. Acad. Sci. USA 91:6574-6578; McQueen-Mason, S., et al., (1992) Plant Cell 4:1425-1433).

Since their first isolation from cucumber hypocotyls, expansin proteins have been identified in many plant species and organs on the basis of activity assays and immunoblotting. Examples include tomato leaves, oat coleoptiles, maize roots, rice internodes, tobacco cell cultures, and various fruits. The original sequencing of cucumber expansin cDNAs has impacted our understanding of expansins in several respects. First, expansin genes have now been identified in many other plant species, and they appear to be restricted largely to the plant kingdom. Second, expansins comprise a large multigene family in the plant species. For example, in Arabidopsis, 31 expansin genes have been identified. Third, studies of expression and localization of expansin mRNA are providing new insights and hypothesis concerning the developmental roles of specific expansin genes. And fourth, sequence comparisons have led to the discovery that another group of proteins known previously as group-1 grass pollen allergens, have expansin activity. These pollen-specific proteins are closely related to a group of sequences known primarily from expressed sequence tag (EST) databases. These EST sequences, together with the group-1 pollen allergens, have now been classified as beta-expansins, whereas the original group of expansins are now classified as alpha-expansins. The α-expansins are described in U.S. Pat. Nos. 5,959,082 and 5,990,283 to Cosgrove et al., which are herein incorporated by reference. β-expansins, in general, are the subject of a previously filed U.S. patent application Ser. No. 09/071,252 filed May 1, 1998. Although these two expansin families have only about 20% amino acid identity, they are similar in size, they share a number of conserved motifs, and they have similar wall-loosening activities.

To date, most studies have focused on α-expansins, and limited work has been done on β-expansins. A soybean cytokinin-induced gene known as CIM1 is now classified as a β-expansin, but the biological function of the CIM1 protein is uncertain. The maize group-1 pollen allergen, Zea m1, has wall-loosening activity with high specificity for grass cell walls. This β-expansin is hypothesized to aid fertilization by loosening the cell walls of the stigma and style, thereby facilitating penetration of the pollen tube. Many other β-expansin sequences are found in the rice EST databases, and most of these sequences come from cDNA libraries made from young seedlings and other plant materials that do not contain pollen. Thus, their biological functions clearly differ from those of the group-1 pollen allergens. These so-called vegetative β-expansins are hypothesized to function in cell enlargement and other processes where wall loosening is required. It is notable that the rice EST collection contains at least 75 entries representing at least 10 distinct β-expansin genes. In contrast, only a single Arabidopsis EST is classified as a β-expansin (although a total of five β-expansin genes are found in the Arabidopsis genome). The disparity in the number of β-expansin entries in the rice and Arabidopsis EST collection, together with the specificity of Zea m1 activity for grass walls, leads to the proposal that β-expansins have evolved specialized function in conjunction with the evolution of the grass cell wall, which has a distinctive set of matrix polysaccharide and structural proteins compared with other land plants. If this is true, one would expect to find an abundance of β-expansin homology in other grasses, with expression in many tissues beside pollen.

Recently, Group 2 and Group 3 allergens (designated group 2/3 allergens or also termed HED2 proteins) have also been shown to have expansin activity. Although these allergens from grass pollen have been studied for many years by immunologists concerned with how they elicit hay fever and related allergic responses in humans, the native activity and biological roles of these proteins have not been examined. Group 2/3 grass pollen allergens are distinguished by pI and immuno-cross reactivity, but accumulating sequence information indicates that they belong to the same protein family, genes for group 2/3 allergens encode a protein with a signal peptide and a mature protein with statistically significant sequence similarity (up to 42% identity) with domain 2 of expansins, with the greatest similarly to group-1 allergen sub-class of β-expansins.

Of the two families of expansins, α- and β-group 2/3 allergens are closest in sequence to the subset β-expansins known to immunologists as the grass pollen group 1 allergens.

Once identified, however, proteins with expansin activity including β-expansins, α-expansins, and group 2/3 allergens, or HED proteins all of which are proteins capable of inducing cell wall extension, have utility not only in the engineered extension of cell walls in living plants but foreseeably in commercial applications where their chemical reactivity. Expansins can disrupt noncovalent associations of cellulose, and as such have particular utility in the paper recycling industry. Paper recycling is a growing concern and will prove more important as the nation's landfill sites become scarcer and more expensive. Paper derives its mechanical strength from hydrogen bonding between paper fibers, which are composed primarily of cellulose. During paper recycling, the hydrogen bonding between paper fibers is disrupted by chemical and mechanical means prior to re-forming new paper products. Proteins which cause cell expansion are thus intrinsically well suited to paper recycling, especially when the proteins are nontoxic and otherwise innocuous, and when the proteins can break down paper products which are resistant to other chemical and enzymatic means of degradation. Use of proteins of this type could thus expand the range of recyclable papers.

Other modes of application of expansins, include production of virgin paper. Pulp for virgin paper is made by disrupting the bonding between plant fibers. For the reasons identified above, expansins are useful in the production of paper pulp from plant tissues. Use of expansins can substitute for harsher chemicals now in use and thereby reduce the financial and environmental costs associated with disposing of these harsh chemicals. The use of expansins can also result in higher quality plant fibers because they would be less degraded than fibers currently obtained by harsher treatments.

Still other modes of applications include the production of ethanol. One of the major limitations and costs associated with ethanol production from cellulose is conversion of cellulose to simple fermentable sugars. Because of the crystalline structure of cellulose, its enzymatic conversion to sugars takes a considerable amount of time and requires large quantities of cellulase enzymes, which are expensive. Likewise for the production of chemically-modified cellulose derivatives, cellulose must be made accessible to reactive chemical agents, this usually requiring high temperature, pressures and harsh chemical conditions. Furthermore, the efficient digestion of straws, hay, and other plant materials by ruminants and other animals is limited by the accessibility of cellulose to the digestive enzymes in the animals' gut. Expansin proteins, particularly, group 2/3/allergans have been shown to made cellulose more easily degraded by cellulase enaymes.

Thus, a continuing need remains for the identification, characterization, and optimization of expansins—proteins which can be characterized as catalysts of the extension of plant cell walls and the weakening of the hydrogen bonds in the pure cellulose.

SUMMARY OF THE INVENTION

The invention relates to crystal structure and activities of Beta-expansins and grass pollen allergens and identification of key regions essential to maximize activity and to identify sequence motifs which correlate with activity.

According to the invention, Beta-expansin structure has been delineated to identify critical regions for activity. For example the β-expansin molecule consists of two domains closely packed and aligned to form a long shallow groove with potential to bind a glycan backbone. The domain has first residues 19-140 which form a protein fold, the second domain includes 147-245 composed of eight β-strands assembled into two anti-parallel sheets. Essential residues include surface aromatic residues W194 and Y160 which are in line with W25 and Y27. From this data one can extrapolate to identify essential regions of conservation to develop modified expansins with improved properties, efficiencies and the like.

DETAILED DESCRIPTION OF THE FIGURES

FIG. 1 is a schematic diagram of the plant cell wall. Cellulose microfibrils are synthesized by large complexes in the plasma membrane and are glued together by branched matrix polysaccharides synthesized in the Golgi and deposited by vesicles along the inner surface of the cell wall. The ˜4 nm wide cellulose microfibril in cross-section consists of ˜36 β-(1→4)-D-glucans organized into a crystalline array. Polysaccharides such as arabinoxylan and xyloglucan spontaneously bind to the surface of cellulose and may also be entrapped during coalescence of the β-(1→4)-D-glucans to form the microfibril. Hydrophilic pectins and structural proteins (not shown) also make up the matrix between cellulose microfibrils and influence the wall's physical properties.

FIG. 2 is a diagram showing the structure of EXPB1 (PDB 2HCZ). A: Ribbon model of EXPB1, showing the overall configuration of the two domains. B: Superposition of the peptide backbone of EXPB1 D1 (shown entirely in red) with the peptide backbone of Humicola Cel45 (PDB code 4ENG), colored green for regions of good alignment with EXPB1, grey otherwise. The yellow residues indicate cellohexaose from the 4ENG model. C: Superposition of residues making up the catalytic site of Humicola Cel45 (blue) and corresponding residues of EXPB1 (red). Other conserved acidic residues this region of EXPB1 are shown in purple. D: Superposition of EXPB1 D2 (colored) and Phl p 2 (grey), a group-2/3 grass pollen allergen (PDB code 1WHO). Coloring scale from best to poorest alignment of peptide backbones: blue-green-yellow-red. E: Top view of the conserved surface of EXPB1, color coded to indicate conservation (red=most conserved; blue=least conserved; white=intermediate). Conserved residues are labelled, and the locations of two antigenic epitopes are indicated (site-D, site-A). F: A model of glucurono-arabinoxylan (yellow and red) was manually fitted to the long open groove of EXPB1 using the program O (66) and subsequently energy minimized using the program CNS (67). Green residues are from D1, cyan residues are from D2 and red residues are the conserved residues identified in panel E. G: End view of same model as in F. Image in E was generated from the program CONSURF (68) using the alignment of 80 EXPB proteins in GenBank and 2HCZ after removal of the N-terminal extension. Images in G and F were generated with PYMOL (DeLano Scientific) after removal of the N-terminal extension.

FIG. 3 is a graph showing the EXPB sequence logo based on 80 EXPB proteins from Genbank, aligned with the sequence of maize EXPB1 (green) and color coded to indicate the structural role of the conserved residues. Residues with unspecified role are indicated in grey. The size of the one-letter amino acid code in the sequence logo indicates the degree of conservation on a logarithmic scale. The logo was generated with the web server at world wide web, weblogo.berkeley.edu. Black lines between Cys residues indicate disulfide bonds.

FIG. 4 shows the hydrolytic activity of expansin B1 against various wall polyaccharides and glycans. A: Hydrolytic activity of EXPB1 against various wall polysaccharides. Data are means ±SEM (n=3). The positive control with arabinoxylan is a crude extract of maize pollen containing endoxylanase activity (69). B: Maize cell walls bind EXPB1. After incubation of EXPB1+/−cell wall, protein remaining in the supernatant was analyzed by SDS-PAGE and stained with SYPRO Ruby. C: EXPB1 binding to isolated polysaccharides immobilized onto nitrocellulose membrane; NC=nitrocellulose membrane along; G=β-(1→3), (1→4)-D-glucan, GM=glucomannan; XG=xyloglucan; OX=oat xylan; BX=birch xylan. Data are means ±SEM (n=3). D: Swelling of maize cell walls after 48-h incubation +/−EXPB1. Methods as described in the binding studies.

FIG. 5 is the amino acid sequence for Zea m 1 isoform d. (Genbank accession number AAO45608).

FIG. 6 is a schematic of the conserved domains of expansin proteins.

DETAILED DESCRIPTION OF THE INVENTION
Background to Crystallization

It is well-known in the art of protein chemistry, that crystallizing a protein is a difficult process. In fact it is now evident that protein crystallization is the main hurdle in protein structure determination. There are many references which describe the difficulties associated with growing protein crystals. For example, Kierzek, A. M. and Zielenkiewicz, P., (2001), Biophysical Chemistry, 91:1-20, Models of protein crystal growth, and Wiencek, J. M. (1999) Annu. Rev. Biomed. Eng., 1:505-534, New Strategies for crystal growth. It is commonly held that crystallization of protein molecules from solution is the major obstacle in the process of determining protein structures. The reasons for this are many; proteins are complex molecules, and the delicate balance involving specific and non-specific interactions with other protein molecules and small molecules in solution is difficult to predict.

Each protein crystallizes under a unique set of conditions which cannot be predicted in advance. Simply supersaturating the protein to bring it out of solution may not work, the result would, in most cases, be an amorphous precipitate. Many precipitating agents are used, common ones are different salts, and polyethylene glycols, but others are known. In addition, additives such as metals and detergents can be added to modulate the behavior of the protein in solution. Many kits are available (e.g. from Hampton Research), which attempt to cover as many parameters in crystallization space as possible, but in many cases these are just a starting point to optimize crystalline precipitates and crystals which are unsuitable for diffraction analysis. Successful crystallization is aided by a knowledge of the proteins behavior in terms of solubility, dependence on metal ions for correct folding or activity, interactions with other molecules and any other information that is available. Even so, crystallization of proteins is often regarded as a time-consuming process, whereby subsequent experiments build on observations of past trials.

In cases where protein crystals are obtained, these are not necessarily always suitable for diffraction analysis; they may be limited in resolution, and it may subsequently be difficult to improve them to the point at which they will diffract to the resolution required for analysis. Limited resolution in a crystal can be due to several things. It may be due to intrinsic mobility of the protein within the crystal, which can be difficult to overcome, even with other crystal forms. It may be due to high solvent content within the crystal, which consequently results in weak scattering. Alternatively, it could be due to defects within the crystal lattice which mean that the diffracted x-rays will not be completely in phase from unit to unit within the lattice. Any one of these or a combination of these could mean that the crystals are not suitable for structure determination.

Some proteins never crystallize, and after a reasonable attempt it is necessary to examine the protein itself and consider whether it is possible to make individual domains, different N or C-terminal truncations, or point mutations. It is often hard to predict how a protein could be re-engineered in such a manner as to improve crystallizability. Our understanding of crystallization mechanisms are still incomplete and the factors of protein structure which are involved in crystallization are poorly understood.

Determination of Protein Structure.

A mathematical operation termed a Fourier transform relates the diffraction pattern observed from a crystal and the molecular structure of the protein comprising the crystal. A Fourier transform may be considered to be a summation of sine and cosine waves each with a defined amplitude and phase. Thus, in theory, it is possible to calculate the electron density associated with a protein structure by carrying out an inverse Fourier transform on the diffraction data. This, however, requires amplitude and phase information to be extracted from the diffraction data. Amplitude information may be obtained by analyzing the intensities of the spots within a diffraction pattern. Current technologies for generating x-rays and recording diffraction data lead to loss of all phase information. This “phase information” must be in some way recovered and the loss of this information represents the “crystallographic phase problem”. The phase information necessary for carrying out the inverse Fourier transform can be obtained via a variety of methods. If a protein structure exists a set of theoretical amplitudes and phases may be calculated using the protein model and then the theoretical phases combined with the experimentally derived amplitudes. An electron density map may then be calculated and the protein structure observed.

If there is no known structure of the protein then alternative methods for obtaining phases must be explored. One method is multiple isomorphous replacement (MIR). This relies on soaking “heavy atom” (i.e. platinum, uranium, mercury, etc) compounds into the crystals and observing how their incorporation into the crystals modifies the spot intensities observed in the diffraction pattern. This method relies on the heavy atoms being incorporated into the protein at a finite number of defined sites. It is a pre-requisite of an isomorphous replacement experiment that the heavy atom soaked crystals remain isomorphous. That is, there should be no appreciable alterations in the physical characteristics of the protein crystal (i.e. perturbations to crystallographic cell dimensions, or significant loss of resolution). Perturbations to the physical properties of the crystal are termed non-isomorphisms and prevent this type of experiment being successfully completed. Successful isomorphous incorporation of heavy atoms into a protein crystal results in the intensities of the spots within the diffraction pattern obtained from the crystal being modified, as compared to the data collected from an identical, unsoaked, (native) crystal. The diffraction data obtained from a successful isomorphous replacement experiment are termed a “derivative” dataset. By mathematically analyzing the “native” and “derivative” datasets it is possible to extract preliminary phase information from the datasets. This phase information, when combined with the experimentally obtained amplitudes from the native dataset, enables an electron density map of the unknown protein molecule to be calculated using the Fourier transform method.

An alternative method for obtaining phase information for a protein of unknown structure is to perform a multi-wavelength anomalous dispersion (MAD) experiment. This relies on the absorption of X-rays by electrons at certain characteristic X-ray wavelengths. Different elements have different characteristic absorption edges. Anomalous scattering by atoms within a protein will modify the diffraction pattern obtained from the protein crystal. Thus if a protein contains atoms which are capable of anomalous scattering a diffraction dataset (anomalous dataset) may be collected at an X-ray wavelength at which this anomalous scattering is maximal. By altering the X-ray wavelength to a value at which there is no anomalous scattering a native dataset may then be collected. Similarly to the MIR case, by mathematically processing the anomalous and native datasets the phase information necessary for the calculation of an electron density map may be determined. The most usual way to introduce anomalous scatterers into a protein is to replace the sulphur containing methionine amino acid residues with selenium containing seleno-methionine residues. This is done by generating recombinant protein that is isolated from cells grown on growth media that contain seleno-methionine. Selenium is capable of anomalously scattering X-rays and may thus be used for a MAD experiment. Further methods for phase determination such as single isomorphous replacement (SIR), single isomorphous replacement anomalous scattering (SIRAS) and direct methods exist, but the principles behind them are similar to MIR and MAD.

The final method generally available for the calculation of the phases necessary for the determination of an unknown protein structure is molecular replacement. This method relies upon the assumption that proteins with similar amino acid sequences (primary sequences) will have a similar fold and three-dimensional structure (tertiary structure). Proteins related by amino acid sequence are termed homologous proteins. If an X-ray diffraction dataset has been collected from a crystal whose protein structure is not known, but a structure has been determined for a homologous protein, then molecular replacement can be attempted. Molecular replacement is a mathematical process that attempts to correlate the dataset obtained from a new protein crystal with the theoretical diffraction pattern calculated for a protein of known structure. If the correlation is sufficiently high some phase information can be extracted from the known protein structure and combined with the amplitudes obtained from the new protein dataset. This enables calculation of a preliminary electron density map for the protein of unknown structure.

If an electron density map has been calculated for a protein of unknown structure then the amino acids comprising the protein must be fitted into the electron density for the protein. This is normally done manually, although high resolution data may enable automatic model building. The process of model building and fitting the amino acids to the electron density can be both a time consuming and laborious process. Once the amino acids have been fitted to the electron density it is necessary to refine the structure. Refinement attempts to maximize the correlation between the experimentally calculated electron density and the electron density calculated from the protein model built. Refinement also attempts to optimize the geometry and disposition of the atoms and amino acids within the user-constructed model of the protein structure. Sometimes manual re-building of the structure will be required to release the structure from local energetic minima. There are now several software packages available that enable an experimentalist to carry out refinement of a protein structure. There are certain geometry and correlation diagnostics that are used to monitor the progress of a refinement. These diagnostic parameters are monitored and rebuilding/refinement continued until the experimenter is satisfied that the structure has been adequately refined.

The present invention relates to the crystal structure of EXPB1 (Genbank accession AA045608; PDB accession 2HCZ), which allows the binding location of the polysaccharides to the compound and its activities to be investigated and determined.

Thus in one aspect, the invention provides a three dimensional structure of EXPB1 set out in FIGS. 2 and 3, and uses, described further herein below of the three dimensional structure.

According to the invention, EXPB1 contains two domains (residues 19-140 [D1] and 147-245 [D2]) connected by a short linker (residues 141-146) and aligned end to end so as to make a closely-packed irregular cylinder ˜66 Å long and 26 Å in diameter (FIG. 2A).

The two EXPB1 domains pack close to one another, making contact via H-bonds and salt bridges between basic residues (K65 and R137) in D1 and acidic residues (E217 and D171) in D2. These residues are highly conserved in the EXPB family (see annotated sequence logo in FIG. 3). Additional hydrogen bonding is found between S72 and D173, as well as between the peptide backbone for C42 and A196. The two domains also make contact via a hydrophobic patch consisting of I44, P51, Y52 and Y92 in D1 and L164, Y167 and the hydrocarbon chain of K166 in D2, residues that are mostly well conserved or have conservative substitutions in the EXPB family. Moreover, six highly conserved glycine residues (G43, 67, 69, 71, 172, 195) are found at the surfaces where the two domains make contact. The lack of side chains in the glycine residues permits close packing of the two domains.

The two EXPB1 domains align so as to form a long, shallow groove with highly conserved polar and aromatic residues suitably positioned to bind a twisted polysaccharide chain of 10 xylose residues (FIGS. 2E-G). The groove extends from the conserved G129 at one end of D1, spans across a stretch of conserved residues in D1 and D2 (see numbered residues in FIG. 2E as well as annotated sequence logo in FIG. 3) and ends at N157, a distance of some 47 Å. Many of the conserved residues common to EXPA and EXPB make up this potential binding surface, including residues in the classic expansin motifs TWYG, GGACG, and HFD (see FIG. 3).

Residues that could bind a polysaccharide by van der Waals interactions with the sugar rings include W26, Y27, G40, and G44 from D1 as well as Y160 and W194 from D2. Conserved residues that might stabilize polysaccharide binding by H-bonding include T25, D37, D95 and D107 in D1 and N157, S193 and R199 in D2.

In general aspects, the present invention is concerned with the provision of an EXPB1 structure and its use in modeling the interaction of molecular structures, e.g. potential and existing substrates, inhibitors, analogs, or fragments of such compounds, with this EXPB1 structure.

These and other aspects and embodiments of the present invention are discussed below. The above aspects of the invention, both singly and in combination, all contribute to features of the invention, which are advantageous.

The invention comprises in one paragraph a computer-based method for the analysis of the interaction of a molecular structure with an EXPB1 structure, which comprises: providing a structure comprising a three-dimensional representation of EXPB1 or a portion thereof, which representation comprises all or a portion of the coordinates of any one of figures represented in FIGS. 2 and 3 providing a molecular structure to be fitted to said EXPB1 structure or selected coordinates thereof; and fitting the molecular structure to said EXPB1 structure.

The method of the invention further comprises the steps of obtaining or synthesizing a compound which has said molecular structure; and contacting said compound with EXPB1 protein to determine the ability of said compound to interact with the EXPB1.

The method also include obtaining or synthesizing a compound which has said molecular structure; forming a complex of an EXPB1 substrate protein and said compound; and analyzing said complex by X-ray crystallography to determine the ability of said compound to interact with the EXPB1 substrate.

The method further comprises the steps of: obtaining or synthesizing a compound which has said molecular structure; and determining or predicting how said compound interacts with an EXPB1 substrate; and modifying the compound structure so as to alter the interaction between it and the substrate. The invention also includes a compound having the modified structure identified using the method and which has expansin activity.

A method of obtaining a structure of a target EXPB1 protein of unknown structure, the method comprises the steps of: providing a crystal of said target EXPB1 protein, obtaining an X-ray diffraction pattern of said crystal, calculating a three-dimensional atomic coordinate structure of said target, by modeling the structure of said target EXPB1 protein of unknown structure on the active site structure of any one of FIGS. 2-3.

The invention also includes methods where the molecular structure to be fitted is in the form of a model of a pharmacophore including but not limited to: (a) a wire-frame model; (b) a chicken-wire model; (c) a ball-and-stick model; (d) a space-filling model; (e) a stick-model; (f) a ribbon model; (g) a snake model; (h) an arrow and cylinder model; (i) an electron density map; (j) a molecular surface model.

The invention also includes a computer-based method for the analysis of molecular structures which comprises: (a) providing the coordinates of at least two atoms of an EXPB1 structure as defined in FIGS. 2 and/or 3 (b) providing the structure of a molecular structure to be fitted to the selected coordinates; and (c) fitting the structure to the selected coordinates of the EXPB1 structure. The method further contemplates that the coordinates will be a at least a portion of a binding pocket.

A computer-based method of protein design comprising: (a) providing the coordinates of at least two atoms of an EXPB1 structure as defined in any one of FIGS. 2 and 3 with a square deviation of less than 1.5 Å (“selected coordinates”); (b) providing the structures of a plurality of EXPB1 substrates or potential substrates; (c) fitting the structure of each of the EXPB1 substrates or potential substrates to the selected coordinates; and (d) determining the activity of said EXPB1 structure on said substrate or potential substrate.

A method for identifying a candidate modulator of EXPB1 comprising the steps of: (a) employing a three-dimensional structure of EXPB1, at least one sub-domain thereof, or a plurality of atoms thereof, to characterize at least one EXPB1 binding cavity, the three-dimensional structure being defined by FIGS. 2-3; and (b) identifying the candidate modulator by designing or selecting a compound for interaction with the binding cavity. The method further comprising the step of: (a) obtaining or synthesizing the candidate modulator; and (b) contacting the candidate modulator with EXPB1 to determine the ability of the candidate modulator to interact with EXPB1.

The invention also contemplates a method for determining the structure of a protein, which method comprises: providing the co-ordinates per FIGS. 2-3 or selected coordinates thereof, and either (a) positioning said co-ordinates in the crystal unit cell of said protein so as to provide a structure for said protein, or (b) assigning NMR spectra peaks of said protein by manipulating said co-ordinates.

A method for determining the structure of a compound bound to EXPB1 protein, said method comprising: providing a crystal of EXPB1 protein; soaking the crystal with the compound to form a complex; and determining the structure of the complex by employing the data of any one of FIGS. 2-3 or a portion thereof.

A method for determining the structure of a compound bound to EXPB1 protein, said method comprising: mixing EXPB1 protein with the compound; crystallizing an EXPB1 protein-compound complex; and determining the structure of the complex by employing the data of any one of Tables 1 or FIGS. 2-3 or a portion thereof.

A method for modifying the structure of a compound in order to alter its metabolism by an EXPB1, which method comprises: fitting a starting compound to one or more coordinates of at least one amino acid residue of the ligand-binding region of the EXPB1; modifying the starting compound structure so as to increase or decrease its interaction with the ligand-binding region.

A method for modifying the structure of a compound in order to alter its metabolism by an EXPB1, which method comprises: fitting a starting compound to one or more coordinates of at least one amino acid residue of the binding region of the EXPB1; modifying the starting compound structure so as to increase or decrease its interaction with the binding region.

A method for modifying the structure of a compound in order to alter its, or another compounds, metabolism by an EXPB1, which method comprises: fitting a starting compound to one or more coordinates of at least one amino acid residue of the peripheral binding region of the EXPB1; modifying the starting compound structure so as to increase or decrease its interaction with the peripheral binding region; wherein said peripheral binding region is defined as the EXPB1 residues numbered as: W26, Y27, G40, nd G44, Y160, and W194.

A method of obtaining a representation of the three dimensional structure of a crystal of EXPB1, which method comprises providing the data of any one of PDB accession #2HCZ or FIGS. 2-3 or selected coordinates thereof, and constructing a three-dimensional structure representing said coordinates.

A computer system, intended to generate structures and/or perform optimization of compounds which interact with EXPB1, EXPB1 homologues or analogues, complexes of EXPB1 with compounds, or complexes of EXPB1 homologues or analogues with compounds, the system containing computer-readable data comprising one or more of: (a) EXPB1 co-ordinate data of any one of PDB accession #2HCZ, of FIGS. 2-3, said data defining the three-dimensional structure of EXPB1 or at least selected coordinates thereof; (b) atomic coordinate data of a target EXPB1 protein generated by homology modeling of the target based on the coordinate data of any one of PDB accession #2HCZ, FIGS. 2-3 (c) atomic coordinate data of a target EXPB1 protein generated by interpreting X-ray crystallographic data or NMR data by reference to the co-ordinate data of any one of PDB accession #2HCZ, or FIGS. 2-3 (d) structure factor data derivable from the atomic coordinate data of (b) or (c). and (e) atomic coordinate data of any one of PDB accession #2HCZ, or FIGS. 2-3 or selected coordinates thereof.

A computer system according to paragraph comprising: (i) a computer-readable data storage medium comprising data storage material encoded with said computer-readable data; (ii) a working memory for storing instructions for processing said computer-readable data; and (iii) a central-processing unit coupled to said working memory and to said computer-readable data storage medium for processing said computer-readable data and thereby generating structures and/or performing rational compound design.

A computer system comprising a display coupled to said central-processing unit for displaying said structures.

A method of providing data for generating structures and/or performing optimization of compounds which interact with EXPB1, EXPB1 homologues or analogues; complexes of EXPB1 with compounds, or complexes of EXPB1 homologues or analogues with compounds, the method comprising: (i) establishing communication with a remote device containing (a) computer-readable data comprising atomic coordinate data of any one of Tables 1, or FIGS. 2-3 or selected coordinates thereof; (b) atomic coordinate data of a target EXPB1 homologue or analogue generated by homology modeling of the target based on the data (a); (c) atomic coordinate data of a protein generated by interpreting X-ray crystallographic data or NMR data by reference to the data of any one of PDB accession #2HCZ, or FIGS. 2-3 and (d) structure factor data derivable from the atomic coordinate data of (d) or (e); and (ii) receiving said computer-readable data from said remote device.

A computer-readable storage medium, comprising a data storage material encoded with computer readable data, wherein the data are defined by all or a portion of the structure coordinates of the EXPB1 protein of any one of PDB accession #2HCZ or FIGS. 2-3 or a homologue of EXPB1, wherein said homologue comprises backbone atoms that have a root mean square deviation from the backbone atoms of said any one of PDB accession #2HCZ, or table 1, or FIGS. 2-3 respectively of not more than 1.5 Å.

A. Protein Crystals.

The present invention provides a crystal of EXPB1 having cell dimensions of about a=113.7 Å, b=45.2 Å and c=70.3 Å. With angles α=90.0°, β=124.6°, and γ=90.0. Unit cell variability of 5% may be observed in all dimensions.

Substrates include plant cell walls, or components thereof. Alternatively the ligand could be a compound whose interaction with EXPB1 is unknown.

Such crystals may be obtained using the methods described in the accompanying examples.

The EXPB1 may optionally comprise a tag, such as a C-terminal polyhistidine tag to allow for recovery and purification of the protein.

The methodology used to provide an EXPB1 crystal illustrated herein may be used generally to provide an EXPB1 crystal resolvable at a resolution of at least 3.0 Å and preferably at least 2.8 Å. The invention thus further provides an EXPB1 crystal having a resolution of at least 3.0 Å, preferably at least 2.8 Å. The proteins may be wild-type proteins or variants thereof, which are modified to promote crystal formation, for example by N-terminal truncations and/or deletion of loop regions, which prevent crystal formation.

In a further aspect, the invention provides a method for making an EXPB1 protein crystal, particularly of an EXPB1 protein comprising the core sequence of EXPB1 (as defined above) or a variant thereof, which method comprises growing a crystal by vapor diffusion using a reservoir buffer that contains 0.05-0.2 M HEPES pH 7.0-7.8, 2.5-10% IPA, 0-20% PEG 4000, 0-0.3 M sodium chloride, 0-10% PEG 400, 0-10% glycerol, preferably 0.1 M HEPES pH 7.2, 5% IPA, 10% PEG 4000. The crystal is grown by vapor diffusion and is performed by placing an aliquot of the solution on a cover slip as a hanging drop above a well containing the reservoir buffer. The concentration of the protein solution used was 0.3-0.7 mM.

Crystals of the invention also include crystals of EXPB1 mutants, chimeras, homologues in the expansin family (e.g. α-expansins, β-expansins, group 2/3 allergens, etc) and alleles.

(i) Mutants

A mutant is an EXPB1 protein characterized by the replacement or deletion of at least one amino acid from the wild type EXPB1. Such a mutant may be prepared for example by site-specific mutagenesis, or incorporation of natural or unnatural amino acids.

The present invention contemplates “mutants” wherein a “mutant” refers to a polypeptide which is obtained by replacing at least one amino acid residue in a native or synthetic EXPB1 with a different amino acid residue and/or by adding and/or deleting amino acid residues within the native polypeptide or at the N- and/or C-terminus of a polypeptide corresponding to EXPB1, and which has substantially the same three-dimensional structure as EXPB1 from which it is derived. By having substantially the same three-dimensional structure is meant having a set of atomic structure co-ordinates that have a root mean square deviation (r.m.s.d.) of less than or equal to about 2.0 Å (preferably less than 1.55 or 1.5 Å, more preferably less than 1.0 Å, and most preferably less than 0.5 Å) when superimposed with the atomic structure co-ordinates of the EXPB1 from which the mutant is derived when at least about 50% to 100% of the C_α atoms of the EXPB1 are included in the superposition. A mutant may have, but need not have, enzymatic or catalytic activity.

To produce homologues or mutants, amino acids present in the said protein can be replaced by other amino acids having similar properties, for example hydrophobicity, hydrophobic moment, antigenicity, propensity to form or break α-helical or β-sheet structures, and so on. Substitutional variants of a protein are those in which at least one amino acid in the protein sequence has been removed and a different residue inserted in its place. Amino acid substitutions are typically of single residues but may be clustered depending on functional constraints e.g. at a crystal contact. Preferably amino acid substitutions will comprise conservative amino acid substitutions. Insertional amino acid variants are those in which one or more amino acids are introduced. This can be amino-terminal and/or carboxy-terminal fusion as well as intrasequence. Examples of amino-terminal and/or carboxy-terminal fusions are affinity tags, MBP tag, and epitope tags.

Amino acid substitutions, deletions and additions which do not significantly interfere with the three-dimensional structure of the EXPB1 will depend, in part, on the region of the EXPB1 where the substitution, addition or deletion occurs. In highly variable regions of the molecule, non-conservative substitutions as well as conservative substitutions may be tolerated without significantly disrupting the three-dimensional structure of the molecule. In highly conserved regions, or regions containing significant secondary structure, conservative amino acid substitutions are preferred.

Conservative amino acid substitutions are well-known in the art, and include substitutions made on the basis of similarity in polarity, charge, solubility, hydrophobicity, hydrophilicity and/or the amphipathic nature of the amino acid residues involved. For example, negatively charged amino acids include aspartic acid and glutamic acid; positively charged amino acids include lysine and arginine; amino acids with uncharged polar head groups having similar hydrophilicity values include the following: leucine, isoleucine, valine; glycine, alanine; asparagine, glutamine; serine, threonine; phenylalanine, tyrosine. Other conservative amino acid substitutions are well known in the art.

In some instances, it may be particularly advantageous or convenient to substitute, delete and/or add amino acid residues in order to provide convenient cloning sites in the cDNA encoding the polypeptide, to aid in purification of the polypeptide, etc. Such substitutions, deletions and/or additions which do not substantially alter the three dimensional structure of EXPB1 will be apparent to those having skills in the art.

It should be noted that the mutants contemplated herein need not exhibit enzymatic activity. Indeed, amino acid substitutions, additions or deletions that interfere with the catalytic activity of the EXPB1 but which do not significantly alter the three-dimensional structure of the catalytic region are specifically contemplated by the invention. Such crystalline polypeptides, or the atomic structure co-ordinates obtained there from, can be used to identify compounds that bind to the protein.

The residues for mutation could easily be identified by those skilled in the art and these mutations can be introduced by site-directed mutagenesis e.g. using a Stratagene QuikChange™ Site-Directed Mutagenesis Kit or cassette mutagenesis methods (see e.g. Ausubel et al., eds., Current Protocols in Molecular Biology, John Wiley & Sons, Inc., New York, and Sambrook et al., Molecular Cloning: a Laboratory Manual, 2nd ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., (1989)).

(ii) Alleles

The present invention contemplates “alleles” wherein allele is used for two or more alternative forms of a gene resulting in different gene products and thus different phenotypes. An allele contains nucleotide changes that have been shown to affect transcription, splicing, translation, post-transcriptional or post-translational modifications or result in at least one amino acid change. These different alleles are particularly important in EXPB1s as some may confer different properties on cell wall expansion onto the phenotype. Alleles are often only different by one or two amino acids.

To the extent that the present invention relates to EXPB1-ligand complexes and mutant, homologue, analogue, allelic form, species variant proteins of EXPB1, crystals of such proteins may be formed. The skilled person would recognize that the conditions provided herein for crystallizing EXPB1 may be used to form such crystals. Alternatively, the skilled person would use the conditions as a basis for identifying modified conditions for forming the crystals.

Thus the aspects of the invention relating to crystals of EXPB1, may be extended to crystals of mutant and mutants of EXPB1 which result in homologue, allelic form, and species variant.

(iii) Crystallization of EXPB1

To produce crystals of EXPB1 protein the final protein is, conveniently, concentrated to 10-60, e.g. 20-40 mg/ml in 10-100 mM potassium phosphate with high salt (e.g. 500 mM NaCl or KCl), optionally also with about 1 mM EDTA and/or about 2 mM dithiothreitol, by using concentration devices which are commercially available. Crystallization of the protein is set up by the 0.5-2/1 hanging or sitting drop methods and the protein is crystallized by vapor diffusion at 5-25° C. against a range of vapor diffusion buffer compositions. It is customary to use a 1:1 ratio of protein solution and vapor diffusion buffer in the hanging drop, and this has been used herein unless stated to the contrary.

Typically the vapor diffusion buffer comprises 0-27.5%, preferably 2.5-27.5% PEG 1K-20 K, preferably 1-8K or PEG 2000MME-5000MME, preferably PEG 2000 MME, or 0-10% Jeffamine M-600 and/or 5-20%, e.g. 10-20% propanol or 15-20% ethanol or about 15%-30%, e.g. about 15% 2-methyl-2,4-pentanediol (MPD), optionally with 0.01 M-1.6 M salt or salts and/or 0-0.15, e.g. 0-0.1 M of a solution buffer and/or 0-35%, such as 0-15%, glycerol and/or 0-35% PEG300-400; but preferably: 10-25% PEG 1K-8K or PEG 2000MME or 0-10% Jeffamine M-600 and/or 5-15%, e.g. 10-15%, propanol or ethanol, optionally with 0.1 M-0.2 M salt or salts and/or 0-0.15, e.g. 0-0.1 M solution buffer and/or PEG400, but more preferably: 15-20% PEG 3350 or PEG 4000 or PEG 2000MME or 0-10% Jeffamine M-600 or 5-15%, e.g. 10-15% propanol or ethanol, optionally with 0.1 M-0.2 M salt or salts and/or 0-0.15 M solution buffer.

Alternatively the vapor diffusion buffer may be 0.1 M HEPES pH 7.5 0.2-0.3 M potassium chloride, 1-5% MPD, 7-14.0% PEG 3350 or PEG 4000, 25-50 mM calcium chloride more specifically 0.1 M HEPES pH 7.5, 0.20-0.30 M KCl, 10-14% PEG 4000, 5% MPD, 25 mM calcium chloride.

The salt may be an alkali metal (particularly lithium, sodium and potassium), alkaline earth metal (e.g. magnesium or calcium), ammonium, ferric, ferrous or transition metal salt (e.g. zinc) of a halide (e.g. bromide, chloride or fluoride), acetate, formate, nitrate, sulfate, tartrate, citrate or phosphate. This includes sodium fluoride, potassium fluoride, ammonium fluoride, ammonium acetate, lithium acetate, magnesium acetate, sodium acetate, potassium acetate, calcium acetate, zinc acetate, ammonium chloride, lithium chloride, magnesium chloride, potassium chloride, sodium chloride, potassium bromide, magnesium formate, sodium formate, potassium formate, ammonium formate, ammonium nitrate, lithium nitrate, potassium nitrate, sodium nitrate, ammonium sulfate, potassium sulfate, lithium sulfate, sodium sulfate, di-sodium tartrate, potassium sodium tartrate, di-ammonium tartrate, potassium dihydrogen phosphate, tri-sodium citrate, tri-potassium citrate, zinc acetate, ferric chloride, calcium chloride, magnesium nitrate, magnesium sulfate, sodium dihydrogen phosphate, di-sodium hydrogen phosphate, di-potassium hydrogen phosphate, ammonium dihydrogen phosphate, di-ammonium hydrogen phosphate, tri-lithium citrate, nickel chloride, ammonium iodide, di-ammonium hydrogen citrate.

Solution buffers if present include, for example, Hepes, Tris, imidazole, cacodylate, tri-sodium citrate/citric acid, tri-sodium citrate/HCl, acetic acid/sodium acetate, phosphate-citrate, sodium potassium phosphate, 2-(N-morpholino)-ethane sulphonic acid/NaOH (MES), CHES or bis-trispropane. The pH range is desirably maintained at pH 4.2-8.5, preferably 4.7-8.5. Solution buffers if present can also include, for example, bicine, bis-tris, CAPS, MOPS, ADA which allow the pH to be maintained in the range 5.8-11.

Crystals may be prepared using a Hampton Research Screening kits, Poly-ethylene glycol (PEG)/ion screens, PEG grid, Ammonium sulphate grid, PEG/ammonium sulphate grid or the like. Crystallization may also be performed in the presence of an inhibitor of EXPB1, e.g. fluvoxamine or 2-phenyl imidazole. EXPB1 crystallization may also be performed in the presence of one or more inhibitors e.g. ketoconazole, metyrapone, fluconazole or triadimefon and/or in the presence of one or more substrate(s) e.g. testosterone or progesterone.

Additives can be added to a crystallization condition identified to influence crystallization. Additive Screens are to be used during the optimization of preliminary crystallization conditions where the presence of additives may assist in the crystallization of the sample and the additives may improve the quality of the crystal e.g. Hampton Research additive screens which use glycerol, polyols and other protein stabilizing agents in protein crystallization (R. Sousa. Acta. Cryst. (1995) D51, 271-277) or divalent cations (Trakhanov, S. and Quiocho, F. A. Protein Science (1995) 4, 9, 1914-1919).

In addition, detergents may be added to a crystallization condition to improve the crystallization behavior e.g. the ionic, non-ionic and zwitterionic detergents found in the Hampton Research detergent screens (McPherson, A., et al., The effects of neutral detergents on the crystallization of soluble proteins, J. Crystal Growth (1986) 76, 547-553).

Alternatively, the vapor diffusion buffer typically comprises 0-27.5% PEG 1K-20 K, preferably 1-8K or PEG 2000MME-5000MME, preferably PEG 2000 MME, or 0-10% Jeffamine M-600 and/or 1-20%, e.g. 1-20% propanol or 15-20% ethanol or about 1%-30%, e.g. about 2-25% 2-methyl-2,4-pentanediol (MPD), optionally with 0.01 M-1.6 M salt or salts and/or 0-0.15 M, e.g. 0-0.1 M, of a solution buffer and/or 0-35%, such as 0-15%, glycerol and/or 0-35% PEG300-400; but preferably: 0-27.5%, preferably 2.5-27.5% PEG 1K-20 K, most preferably 5-20% PEG 4K or PEG 2000MME-5000MME, preferably PEG 2000 MME, and 1-20% alcohol, e.g. 1-20% propanol e.g. iso-propanol or 2-25% 2-methyl-2,4-pentanediol (MPD), optionally with 0.01 M-1.6 M salt or salts and/or 0-0.15 M, e.g. 0-0.1 M, of a solution buffer and/or 0-35%, such as 0-15%, glycerol and/or 0-35% PEG300-400.

B. Crystal Coordinates.

In a further aspect, the invention also provides a crystal of EXPB1 having the three dimensional atomic coordinates of PDB accession #2HCZ, the description herein, table 1, and/or FIGS. 2-3.

Protein structure similarity is routinely expressed and measured by the root mean square deviation (r.m.s.d.), which measures the difference in positioning in space between two sets of atoms. The r.m.s.d. measures distance between equivalent atoms after their optimal superposition. The r.m.s.d. can be calculated over all atoms, over residue backbone atoms (i.e. the nitrogen-carbon-carbon backbone atoms of the protein amino acid residues), main chain atoms only (i.e. the nitrogen-carbon-oxygen-carbon backbone atoms of the protein amino acid residues), side chain atoms only or more usually over C-α atoms only. For the purposes of this invention, the r.m.s.d. can be calculated over any of these, using any of the methods outlined below.

Thus the coordinates disclosed herein provide a measure of atomic location in Angstroms, given to 3 decimal places. The coordinates are a relative set of positions that define a shape in three dimensions, but the skilled person would understand that an entirely different set of coordinates having a different origin and/or axes could define a similar or identical shape. Furthermore, the skilled person would understand that varying the relative atomic positions of the atoms of the structure so that the root mean square deviation of the residue backbone atoms (i.e. the nitrogen-carbon-carbon backbone atoms of the protein amino acid residues) is less than 2.0 Å, preferably less than 1.55 or 1.5 Å, more preferably less than 1.0 Å, more preferably less than 0.5 Å, more preferably less than 0.3 Å, such as less than 0.25 Å, or less than 0.2 Å, and most preferably less than 0.1 Å, when superimposed on the coordinates provided in PDB accession #2HCZ for the residue backbone atoms, will generally result in a structure which is substantially the same as the structures disclosed herein in terms of both its structural characteristics and usefulness for structure-based analysis of EXPB1-interactivity molecular structures.

A further rmsd value of less than 1.0 Å which is preferred is a value of less than 0.6 Å, and rmsd values of less than 0.5 Å which are preferred are values of less than 0.45 Å, preferably less than 0.35 Å.

Unless explicitly set out to the contrary, or otherwise clear from the context, reference throughout the present specification to the use of all or selected coordinates disclosed herein does not exclude the use of additional coordinates.

Methods of comparing protein structures are discussed in Methods of Enzymology, vol 115, pg 397-420. The necessary least-squares algebra to calculate r.m.s.d. has been given by Rossman and Argos (J. Biol. Chem., vol 250, pp 7525 (1975)) although faster methods have been described by Kabsch (Acta Crystallogr., Section A, A92, 922 (1976)); Acta Cryst. A34, 827-828 (1978)), Hendrickson (Acta Crystallogr., Section A, A35, 158 (1979)); McLachan (J. Mol. Biol., vol 128, pp 49 (1979)) and Kearsley (Acta Crystallogr., Section A, A45, 208 (1989)). Some algorithms use an iterative procedure in which the one molecule is moved relative to the other, such as that described by Ferro and Hermans (Ferro and Hermans, Acta Crystallographic, A33, 345-347 (1977)). Other methods e.g. Kabsch's algorithm locate the best fit directly.

Programs for determining rmsd include MNYFIT (part of a collection of programs called COMPOSER, Sutcliffe, M. J., Haneef, I., Carney, D. and Blundell, T. L. (1987) Protein Engineering, 1, 377-384), MAPS (Lu, G. An Approach for Multiple Alignment of Protein Structures (1998, in manuscript and on http://bioinfol.mbfys.lu.se/TOP/maps.html)).

It is usual to consider C-alpha atoms and the rmsd can then be calculated using programs such as LSQKAB (Collaborative Computational Project 4. The CCP4 Suite: Programs for Protein Crystallography, Acta Crystallographica, D50, (1994), 760-763), QUANTA (Jones et al., Acta Crystallography A47 (1991), 110-119 and commercially available from Accelerys, San Diego, Calif.), Insight (commercially available from Accelerys, San Diego, Calif.), Sybyl.®. (commercially available from Tripos, Inc., St Louis), O (Jones et al., Acta Crystallographica, A47, (1991), 110-119), and other coordinate fitting programs.

In, for example the programs LSQKAB and O, the user can define the residues in the two proteins that are to be paired for the purpose of the calculation. Alternatively, the pairing of residues can be determined by generating a sequence alignment of the two proteins, programs for sequence alignment are discussed in more detail in Section F. The atomic coordinates can then be superimposed according to this alignment and an r.m.s.d. value calculated. The program Sequoia (C. M. Bruns, I. Hubatsch, M. Ridderstrom, B. Mannervik, and J. A. Tainer (1999) Human Glutathione Transferase A4-4 Crystal Structures and Mutagenesis Reveal the Basis of High Catalytic Efficiency with Toxic Lipid Peroxidation Products, Journal of Molecular Biology 288(3): 427-439) performs the alignment of homologous protein sequences, and the superposition of homologous protein atomic coordinates. Alternatively, the program Astex-KFIT (published in WO2004/038015) can be used. Once aligned, the r.m.s.d. can be calculated using programs detailed above. For sequence identical, or highly identical, the structural alignment of proteins can be done manually or automatically as outlined above. Another approach would be to generate a superposition of protein atomic coordinates without considering the sequence.

It is more normal when comparing significantly different sets of coordinates to calculate the rmsd value over C-α atoms only. It is particularly useful when analyzing side chain movement to calculate the rmsd over all atoms and this can be done using LSQKAB and other programs.

Those of skill in the art will appreciate that in many applications of the invention, it is not necessary to utilize all the coordinates disclosed herein, but merely a portion of them. For example, as described below, in methods of modeling candidate compounds with EXPB1, selected coordinates of EXPB1 may be used.

By “selected coordinates” it is meant for example at least 5, preferably at least 10, more preferably at least 50 and even more preferably at least 100, for example at least 500 or at least 1000 atoms of the EXPB1 structure. Likewise, the other applications of the invention described herein, including homology modeling and structure solution, and data storage and computer assisted manipulation of the coordinates, may also utilize all or a portion of the coordinates (i.e. selected coordinates). The selected coordinates may include or may consist of atoms found in the EXPB1 binding pocket, as described herein below.

C. Description of Structure.

EXPB1 contains two domains (residues 19-140 [D1] and 147-245 [D2]) connected by a short linker (residues 141-146) and aligned end to end so as to make a closely-packed irregular cylinder ˜66 Å long and 26 Å in diameter (FIG. 2A). At its N-terminus EXPB1 has a flexible sequence (residues 1-18) containing hydroxyproline (O9) and a glycan attached to N10, part of the glycosylation consensus sequence NXT. The end of the glycan comes close to the polysaccharide-binding groove (see below) of the symmetry-related protein in the crystalline lattice, with one of the mannose residues stacking against the planar surface formed by residues Gly39 and Gly40 and stabilized further by two hydrogen bonds with the side chain of D37. These interactions with the symmetry-related protein account in part for the unusual ordering of the glycan, as well as the ability to crystallize the glycosylated protein.

Based on its electron density, our model of this N-linked glycan consists of a (1→4)-linked backbone of GlcNac₁GlcNac₂Man₃with two Man residues and a Xyl residue attached to Man₃and a Fuc residue linked to GlcNac₁(FIG. 5, which is published as supporting information on the PNAS web site). Such so-called paucimannosidic-type N-linked glycans are characteristically processed in the Golgi and in post-Golgi steps (31).

Residues 1-3 in the leader sequence were not modeled due to insufficient electron density, but N-terminal sequencing and mass spectrometry indicate their presence (24). The 24-aa signal peptide at the N-terminus, predicted from the EXPB1 cDNA, was absent and was presumably excised during ER processing prior to secretion. No other post-translational modifications, bound metals or ligands were evident from the crystal structure.

The two EXPB1 domains pack close to one another, making contact via H-bonds and salt bridges between basic residues (K65 and R137) in D1 and acidic residues (E217 and D171) in D2. These residues are highly conserved in the EXPB family (see annotated sequence logo in FIG. 3). Additional hydrogen bonding is found between S72 and D173, as well as between the peptide backbone for C42 and A196. The two domains also make contact via a hydrophobic patch consisting of 144, P51, Y52 and Y92 in D1 and L164, Y167 and the hydrocarbon chain of K166 in D2, residues that are mostly well conserved or have conservative substitutions in the EXPB family. Moreover, six highly conserved glycine residues (G43, 67, 69, 71, 172, 195) are found at the surfaces where the two domains make contact. The lack of side chains in the glycine residues permits close packing of the two domains.

Structure of Domain 1. Residues 19-140 form an irregular ovoid with rough dimensions of 35×30×24 Å. The protein fold is dominated by a six-stranded β-barrel flanked by short loops and α-helices (FIG. 2A). D1 has three disulfide bonds (FIG. 3), and the six participating cysteines are highly conserved in both EXPA and EXPB families.

Previous analysis (2, 3) indicated that D1 has distant sequence similarity to members of glycoside hydrolase family 45 (GH45), whose members have been characterized as inverting endo-β-(1→4)-D-glucanases (2, 3, 32, 33). Superposition of D1 with a GH45 protein (PDB #4ENG) using the secondary structure matching algorithm in CCP4 (34) gives good overlap of the two structures for 84 residues (60%) of the peptide backbone of D1 (FIG. 2B), with an root mean square deviation (rmsd) of 2.5 Å. Two of the three disulfide bonds in D1 superimpose exactly with 4ENG disulfides (the exception being C78-C84). Likewise, all of the β-strands in D1 superimpose on β-strands of 4ENG, although the β-strands in EXPB1 are generally shorter. Both structures have short α-helices, but these do not overlap in the two structures.

The GH45 enzyme is substantially larger than D1 (210 residues versus 121) and the “extra” structure in the GH45 enzyme is composed largely of loop regions and α-helices forming a large ridge and subtending structure lacking in D1 (FIG. 2B). In 4ENG this ridge makes a steep border on one side of the deep glucan-binding cleft. Because this ridge is missing in D1, the corresponding surface is more like an open groove than a deep cleft, with space to bind a large, branched polysaccharide (FIGS. 2F, G).

In addition to partial conservation of the protein fold, D1 has noteworthy, but incomplete, conservation of the catalytic site identified in GH45 enzymes (FIG. 2C). In 4ENG (residues designated with *) the catalytic site is centered on aromatic residue Y8* which binds a glucose residue and is flanked by two acidic residues, D10* and D121*, serving as catalytic base and proton donor, respectively, for hydrolysis of the glycosidic bond (33, 35). D121* is flanked on one side by the hydrophobic side chains of A74* and Y8* and on the other side is part of a hydrogen-bonded network with T6*, which in turn is hydrogen bonded to H119*. In D1, a nearly identical structure is found (FIG. 2C), where D107 corresponds to the proton donor D121*, with C58 and Y27 forming the hydrophobic pocket, while T25 and H105 overlap the corresponding residues in 4ENG. Thus D1 possesses much of the conserved catalytic machinery for glycan hydrolysis.

What is missing in EXPB1 is a residue corresponding to D10*, the catalytic base required for glucan hydrolysis by GH45 enzymes (35). As indicated in FIG. 2C D10* is located on a loop that is not aligned with any part of EXPB1. EXPB proteins do have a conserved acidic residue, D37, which is located in a loop (residues 29-38) in the general vicinity corresponding to D10* in 4ENG. This loop is well resolved in D1. However, D37 is located too far from D107 and Y27 to function as the required base. In 4ENG, the catalytic carboxylate groups are located 8.5 Å apart, which is sufficient distance to accommodate a water molecule needed for hydrolysis (35). In D1, the carboxylates for D107 and D37 are 15 Å apart, too distant for this catalytic mechanism. Moreover, simple lateral movement of the loop to bring D37 into a correct position seems unlikely as the loop residues following D37 are rigidly held in place by a several stabilizing interactions. Thus, a key part of the catalytic machinery required for hydrolytic activity of GH45 enzymes is lacking in EXPB1.

Inspection of the EXPB1 structure revealed another acidic residue, D95, which is close to D107 (the carboxylate groups are 8.5 Å away). D95 is highly conserved in group-1 allergens, as well as in β-expansins in general (FIG. 3), but not in α-expansins. However, D95 is not correctly positioned, relative to the D107/Y27 site and the presumed position of the glycan backbone to serve as the catalytic base for hydrolysis. D95 and D37 have an appropriate distance from each other to potentially serve in hydrolysis of a sugar residue, which might be bound to the planar hydrophobic surface made up of G39, G40 and A41 backbone atoms, but none of these residues are part of the site that is conserved with GH45 enzymes.

Enzymatic activity. Because of the structural similarity between D1 and GH45 and the configuration of D95/D37, we tested the ability of EXPB1 to hydrolyze the major polysaccharides of the cell wall. Even with 48-h incubations, we did not detect hydrolytic activity by EXPB1 (FIG. 4A).

Taking another tack, we tested two GH45 enzymes (32, 36) and a nonenzymatic GH45-related protein named “swollenin” (37) for their abilities to catalyze cell wall extension. For these experiments, heat-inactivated walls from cucumber hypocotyls and wheat coleoptiles were clamped in tension in an extensometer and changes in length were monitored upon addition of protein. We observed only small traces of wall extension activity for the GH45 enzymes and for swollenin. Thus, these related proteins lack significant expansin-type activity, at least with the cell walls tested here.

We conclude that, despite the structural similarity of D1 to GH45, EXPB1 does not induce wall extension via wall polysaccharide hydrolysis.

Structure of Domain 2 (D2). Residues 147-245 of EXPB1 make up a second domain (D2) composed of eight β strands assembled into two antiparallel β sheets (FIG. 2A). The two β sheets are at slight angles to each other and form a β-sandwich similar to the immunoglobulin fold. D2 has 36% sequence identity with Phl p 2, a group-2/3 grass pollen allergen (PDB #1WHO), and superposition of the two structures shows them to have identical folds (rmsd of 1.3 Å; FIG. 2D). In comparing the two structures, we find that D2 tends to have shorter β strands compared with Phl p 2 and the two proteins deviate slightly in the loop regions connecting the β-strands.

Identification and Use of EXPB1 Binding Pocket Residues.

D1 and D2 form a long potential polysaccharide-binding site. The two EXPB1 domains align so as to form a long, shallow groove with highly conserved polar and aromatic residues suitably positioned to bind a twisted polysaccharide chain of 10 xylose residues (FIG. 2E-G). The groove extends from the conserved G129 at one end of D1, spans across a stretch of conserved residues in D1 and D2 (see numbered residues in FIG. 2E as well as annotated sequence logo in FIG. 3) and ends at N157, a distance of some 47 Å. Many of the conserved residues common to EXPA and EXPB make up this potential binding surface, including residues in the classic expansin motifs TWYG, GGACG, and HFD (see FIG. 3).

D. Chimeras.

The use of chimeric proteins to achieve desired properties is now common in the scientific literature. Active site chimeras are also described: for example, Swairjo et al (Biochemistry (1998) 37:10928-10936) made loop chimeras of HIV-1 and HIV-2 protease to try to understand determinants of inhibitor-binding specificity.

Of particular relevance are cases where the active site is modified so as to provide a surrogate system to obtain structural information. Thus Ikuta et al (J Biol Chem (2001) 276:27548-27554) modified the active site of cdk2, for which they could obtain structural data, to resemble that of cdk4, for which no X-ray structure is currently available. In this way they were able to obtain protein/ligand structures from the chimeric protein which were useful in cdk4 inhibitor design. In a similar way, based on comparison of primary sequences of highly related isoforms the active site of the EXPB1 protein could be modified to resemble those isoforms. Protein structures or protein/ligand structures of the chimeric proteins could be used in structure-based alteration of the metabolism of compounds which are substrates of that related EXPB1 isoform.

(i) Converting Other EXPB1 Proteins to EXPB1-Like Chimeras

Aspects of the present invention therefore relate to modification of EXPB1 proteins such that the active sites mimic those of related isoforms. For example, from a knowledge of the structure and residues of the active site of the maize EXPB1 structure contained herein, a person skilled in the art could modify an EXPB1 protein such that the active site mimicked that of maize EXPB1. This protein could then be used to obtain information on compound binding through the determination of protein/ligand complex structures using the chimeric EXPB1 protein.

For example, in one aspect the present invention provides a chimeric protein having a binding cavity which provides a substrate specificity substantially identical to that of EXPB1 protein, wherein the chimeric protein binding cavity is lined by a plurality of atoms which correspond to selected EXPB1 atoms lining the EXPB1 binding cavity, and the relative positions of the plurality of atoms corresponding to the relative positions, as defined herein.

E. Homology Modeling.

The invention also provides a means for homology modeling of other proteins (referred to below as target EXPB1 proteins). By “homology modeling”, it is meant the prediction of related EXPB1 structures based either on X-ray crystallographic data or computer-assisted de novo prediction of structure, based upon manipulation of the coordinate data derivable herein or selected portions thereof.

“Homology modeling” extends to target EXPB1 proteins which are analogues or homologues of the EXPB1 protein whose structure has been determined in the accompanying examples. It also extends to EXPB1 protein mutants of EXPB1 protein itself.

The term “homologous regions” describes amino acid residues in two sequences that are identical or have similar (e.g. aliphatic, aromatic, polar, negatively charged, or positively charged) side-chain chemical groups. Identical and similar residues in homologous regions are sometimes described as being respectively “invariant” and “conserved” by those skilled in the art.

In general, the method involves comparing the amino acid sequences of the EXPB1 protein with a target EXPB1 protein by aligning the amino acid sequences. Amino acids in the sequences are then compared and groups of amino acids that are homologous (conveniently referred to as “corresponding regions”) are grouped together. This method detects conserved regions of the polypeptides and accounts for amino acid insertions or deletions as seen in FIG. 3.

Homology between amino acid sequences can be determined using commercially available algorithms. The programs BLAST, gapped BLAST, BLASTN, PSI-BLAST and BLAST2 (provided by the National Center for Biotechnology Information) are widely used in the art for this purpose, and can align homologous regions of two amino acid sequences. These may be used with default parameters to determine the degree of homology between the amino acid sequence of the protein and other target EXPB1 proteins which are to be modeled.

Analogues are defined as proteins with similar three-dimensional structures and/or functions with little evidence of a common ancestor at a sequence level.

Homologues are defined as proteins with evidence of a common ancestor, i.e. likely to be the result of evolutionary divergence and are divided into remote, medium and close sub-divisions based on the degree (usually expressed as a percentage) of sequence identity.

A homologue is defined here as a protein with at least 15% sequence identity or which has at least one functional domain, which is characteristic of EXPB1. This includes polymorphic forms of EXPB1.

There are two types of homologue: orthologues and paralogues. Orthologues are defined as homologous genes in different organisms, i.e. the genes share a common ancestor coincident with the speciation event that generated them. Paralogues are defined as homologous genes in the same organism derived from a gene/chromosome/genome duplication, i.e. the common ancestor of the genes occurred since the last speciation event.

The homologues could also be polymorphic forms of EXPB1 such as alleles or mutants as described in section (A).

Once the amino acid sequences of the polypeptides with known and unknown structures are aligned, the structures of the conserved amino acids in a computer representation of the polypeptide with known structure are transferred to the corresponding amino acids of the polypeptide whose structure is unknown. For example, a tyrosine in the amino acid sequence of known structure may be replaced by a phenylalanine, the corresponding homologous amino acid in the amino acid sequence of unknown structure.

The structures of amino acids located in non-conserved regions may be assigned manually by using standard peptide geometries or by molecular simulation techniques, such as molecular dynamics. The final step in the process is accomplished by refining the entire structure using molecular dynamics and/or energy minimization.

Homology modeling as such is a technique that is well known to those skilled in the art (see e.g. Greer, Science, Vol. 228:1055 (1985), and Blundell et al., Eur. J. Biochem, Vol. 172:513 (1988)). The techniques described in these references, as well as other homology modeling techniques, generally available in the art, may be used in performing the present invention.

Thus the invention provides a method of homology modeling comprising the steps of: (a) aligning a representation of an amino acid sequence of a target EXPB1 protein of unknown three-dimensional structure with the amino acid sequence of the EXPB1 herein to match homologous regions of the amino acid sequences; (b) modeling the structure of the matched homologous regions of said target EXPB1 of unknown structure on the corresponding regions of the EXPB1 structure as obtained as described above and/or that of any one of Tables 1-4 or selected coordinates thereof; and (c) determining a conformation (e.g. so that favorable interactions are formed within the target EXPB1 of unknown structure and/or so that a low energy conformation is formed) for said target EXPB1 of unknown structure which substantially preserves the structure of said matched homologous regions. Preferably one or all of steps (a) to (c) are performed by computer modeling.

The aspects of the invention described herein which utilize the EXPB1 structure in silico may be equally applied to homologue models of EXPB1 obtained by the above aspect of the invention, and this application forms a further aspect of the present invention. Thus having determined a conformation of an EXPB1 by the method described above, such a conformation may be used in a computer-based method of rational drug design as described herein.

F. Structure Solution

The atomic coordinate data of EXPB1 can also be used to solve the crystal structure of other target EXPB1 proteins including other crystal forms of EXPB1, mutants, co-complexes of EXPB1, where X-ray diffraction data or NMR spectroscopic data of these target EXPB1 proteins has been generated and requires interpretation in order to provide a structure.

In the case of EXPB1, this protein may crystallize in more than one crystal form. The data, as provided by this invention, are particularly useful to solve the structure of those other crystal forms of EXPB1. It may also be used to solve the structure of EXPB1 mutants, EXPB1 co-complexes, or of the crystalline form of any other protein with significant amino acid sequence homology to any functional domain of EXPB1.

In the case of other target EXPB1 proteins, particularly the maize EXPB1 proteins referred to in Section E above, the present invention allows the structures of such targets to be obtained more readily where raw X-ray diffraction data is generated.

Thus, where X-ray crystallographic or NMR spectroscopic data is provided for a target EXPB1 of unknown three-dimensional structure, the atomic coordinate data derived herein, may be used to interpret that data to provide a likely structure for the other EXPB1 by techniques which are well known in the art, e.g. phasing in the case of X-ray crystallography and assisting peak assignments in NMR spectra.

One method that may be employed for these purposes is molecular replacement. In this method, the unknown crystal structure, whether it is another crystal form of EXPB1, an EXPB1 mutant, an EXPB1 chimera or an EXPB1 co-complex, or the crystal of a target EXPB1 protein with amino acid sequence homology to any functional domain of EXPB1, may be determined using the EXPB1 structure coordinates. This method will provide an accurate structural form for the unknown crystal more quickly and efficiently than attempting to determine such information ab initio.

Examples of computer programs known in the art for performing molecular replacement are CNX (Brunger A. T.; Adams P. D.; Rice L. M., Current Opinion in Structural Biology, Volume 8, Issue 5, October 1998, Pages 606-611 (also commercially available from Accelrys San Diego, Calif.), MOLREP (A. Vagin, A. Teplyakov, MOLREP: an automated program for molecular replacement, J. Appl. Cryst. (1997) 30, 1022-1025, part of the CCP4 suite) or AMoRe (Navaza, J. (1994). AMoRe: an automated package for molecular replacement. Acta Cryst. A50, 157-163).

G. Computer Systems.

In another aspect, the present invention provides systems, particularly a computer system, the systems containing one of (a) EXPB1 co-ordinate data herein, said data defining the three-dimensional structure of EXPB1 or at least selected coordinates thereof; (b) atomic coordinate data of a target EXPB1 protein generated by homology modeling of the target based on the coordinate data herein, (c) atomic coordinate data of a target EXPB1 protein generated by interpreting X-ray crystallographic data or NMR data by reference to the co-ordinate data herein; or (d) structure factor data derivable from the atomic coordinate data of (b) or (c).

For example the computer system may comprise: (i) a computer-readable data storage medium comprising data storage material encoded with the computer-readable data; (ii) a working memory for storing instructions for processing said computer-readable data; and (iii) a central-processing unit coupled to said working memory and to said computer-readable data storage medium for processing said computer-readable data and thereby generating structures and/or performing rational compound design. The computer system may further comprise a display coupled to said central-processing unit for displaying said structures.

The invention also provides such systems containing atomic coordinate data of target EXPB1 proteins wherein such data has been generated according to the methods of the invention described herein based on the starting data provided the data herein or selected coordinates thereof.

Such data is useful for a number of purposes, including the generation of structures to analyze the mechanisms of action of EXPB1 proteins and/or to perform rational drug design of compounds, which interact with EXPB1.

In a further aspect, the present invention provides computer readable media with at least one of (a) EXPB1 co-ordinate data herein, said data defining the three-dimensional structure of EXPB1 or at least selected coordinates thereof; (b) atomic coordinate data of a target EXPB1 protein generated by homology modeling of the target based on the coordinate data herein, (c) atomic coordinate data of a target EXPB1 protein generated by interpreting X-ray crystallographic data or NMR data by reference to the co-ordinate data; or (d) structure factor data derivable from the atomic coordinate data of (b) or (c).

In another aspect, the invention provides a computer-readable storage medium, comprising a data storage material encoded with computer readable data, wherein the data are defined by all or a portion (e.g. selected coordinates as defined herein) of the structure coordinates of EXPB1 herein, or a homologue of said EXPB1, wherein said homologue comprises backbone atoms that have a root mean square deviation from the Cα or backbone atoms (nitrogen-carbon_α-carbon) of less than 2 Å, preferably less than 1.55 or 1.5 Å, more preferably less than 1.0 Å (e.g. less than 0.6 Å), and most preferably less than 0.5 Å (e.g. less than 0.45 Å such as less than 0.35 Å).

As used herein, “computer readable media” refers to any medium or media, 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 optical discs or CD-ROM; electrical storage media such as RAM and ROM; and hybrids of these categories such as magnetic/optical storage media.

By providing such computer readable media, the atomic coordinate data of the invention can be routinely accessed to model EXPB1s or selected coordinates thereof. For example, RASMOL (Sayle et al., TIBS, Vol. 20, (1995), 374) is a publicly available computer software package, which allows access and analysis of atomic coordinate data for structure determination and/or rational drug design.

As used herein, “a computer system” refers to the hardware means, software means and data storage means used to analyze the atomic coordinate data of the 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. Desirably a monitor is provided to visualize structure data. The data storage means may be RAM or means for accessing computer readable media of the invention. Examples of such systems are microcomputer workstations available from Silicon Graphics Incorporated and Sun Microsystems running Unix based, Windows NT or IBM OS/2 operating systems.

The invention also provides a computer-readable data storage medium comprising a data storage material encoded with a first set of computer-readable data comprising the EXPB1 coordinates herein or selected coordinates thereof; which, when combined with a second set of machine readable data comprising an X-ray diffraction pattern of a molecule or molecular complex of unknown structure, using a machine programmed with the instructions for using said first set of data and said second set of data, can determine at least a portion of the electron density corresponding to the second set of machine readable data.

H. Uses of the Structures of the Invention.

The crystal structures obtained according to the present invention as well as the structures of target EXPB1 proteins obtained in accordance with the methods described herein), may be used in several ways for chemical compound design.

In the case where a molecule is bound by an EXPB1, information on the binding orientation by either co-crystallization, soaking or computationally docking the binding orientation of the compound in the binding pocket can be determined. This will guide specific modifications to the chemical structure designed to mediate or control the interaction of the compound with the protein. Such modifications can be designed with an aim to increase the enhancement of activity by EXPB1 or to increase the active life of the compound and so improve its enzymatic activity.

The crystal structure could also be useful to understand EXPB1-cellulose (substrate) interactions. The crystal structure of the present invention complexed to such a modulator or other compound (either in vitro or in silico) may also allow rational modifications either to modify the modulator such that it either increases or decreases activity, or to modify the EXPB1 such that it could bind better and so displace the modulator.

EXPB1s, as all expansins display significant polymorphic variations dependent on the plant species. This can manifest itself in adverse reactions from some uses. By using the crystal structures of the present invention to map the relevant mutation with respect to the binding mode of EXPB1, chemical modifications could also be made to the expansin to avoid interactions with the variable region of the protein. This could ensure more consistent polysaccharide binding and cell wall extension from EXPB1 for such segments of the population and avoid unwanted deleterious effects.

Some compounds may be converted by EXPB1s into active metabolites. In the case of such compounds, a greater understanding of how such compounds are converted by an EXPB1 will allow modification of the compound so that it can be converted at a different rate. For example, increasing the rate of conversion may allow a more rapid delivery of a desired wall loosening effect, whereas decreasing the rate of conversion may allow for higher sustained activity.

Thus, the determination of the three-dimensional structure of EXPB1 provides a basis for the design of new compounds, which interact with EXPB1 in novel ways. For example, knowing the three-dimensional structure of EXPB1, computer modeling programs may be used to design different molecules expected to interact with possible or confirmed active sites, such as binding sites or other structural or functional features of EXPB1.

(i) Obtaining and Analyzing Crystal Complexes.

In one approach, the structure of a compound bound to an EXPB1 may be determined by experiment. This will provide a starting point in the analysis of the compound bound to EXPB1, thus providing those of skill in the art with a detailed insight as to how that particular compound interacts with EXPB1 and the mechanism by which it is metabolized.

Many of the techniques and approaches to structure-based compound design described above rely at some stage on X-ray analysis to identify the binding position of a ligand in a ligand-protein complex. A common way of doing this is to perform X-ray crystallography on the complex, produce a difference Fourier electron density map, and associate a particular pattern of electron density with the ligand. However, in order to produce the map (as explained e.g. by Blundell et al., in Protein Crystallography, Academic Press, New York, London and San Francisco, (1976)), it is necessary to know beforehand the protein 3D structure (or at least the protein structure factors). Therefore, determination of the EXPB1 structure also allows difference Fourier electron density maps of EXPB1-compound complexes to be produced, determination of the binding position of the drug and hence may greatly assist the process of rational drug design.

Accordingly, the invention provides a method for determining the structure of a compound bound to EXPB1, said method comprising: providing a crystal of EXPB1 according to the invention; soaking the crystal with said compounds; and determining the structure of said EXPB1 compound complex by employing the data described herein.

Alternatively, the EXPB1 and compound may be co-crystallized. Thus the invention provides a method for determining the structure of a compound bound to EXPB1, said method comprising; mixing the protein with the compound(s), crystallizing the protein-compound(s) complex; and determining the structure of said EXPB1-compound(s) complex by reference to the EXPB1 structural data herein.

The analysis of such structures may employ (i) X-ray crystallographic diffraction data from the complex and (ii) a three-dimensional structure of EXPB1, or at least selected coordinates thereof, to generate a difference Fourier electron density map of the complex, the three-dimensional structure being defined by atomic coordinate data provided herein. The difference Fourier electron density map may then be analyzed.

Therefore, such complexes can be crystallized and analyzed using X-ray diffraction methods, e.g. according to the approach described by Greer et al., J. of Medicinal Chemistry, Vol. 37, (1994), 1035-1054, and difference Fourier electron density maps can be calculated based on X-ray diffraction patterns of soaked or co-crystallized EXPB1 and the resolved structure of uncomplexed EXPB1. These maps can then be analyzed e.g. to determine whether and where a particular compound binds to EXPB1 and/or changes the conformation of EXPB1.

Electron density maps can be calculated using programs such as those from the CCP4 computing package (Collaborative Computational Project 4. The CCP4 Suite: Programs for Protein Crystallography, Acta Crystallographica, D50, (1994), 760-763.). For map visualization and model building programs such as “O” (Jones et al., Acta Crystallographica, A47, (1991), 110-119) can be used.

In addition, in accordance with this invention, EXPB1 mutants may be crystallized in co-complex with known EXPB1 substrates or inhibitors or novel compounds. The crystal structures of a series of such complexes may then be solved by molecular replacement and compared with that of the EXPB1 structure disclosed herein. Potential sites for modification within the various binding sites of the protein may thus be identified. This information provides an additional tool for determining the most efficient binding interactions, for example, increased hydrophobic interactions, between EXPB1 and a chemical entity or compound.

For example there are alleles of EXPB1, which differ from the native EXPB1 by only 1-2 amino acid substitutions, and yet individuals who express these allelic variants may exhibit different binding affinities or activities. The metabolism of enzymatic agents used in the hydrolysis of cellulose or plant cell wall extension applications can be investigated using the structure provided here and the agents then altered using the methods described herein.

This information may thus be used to optimize known classes of EXPB1 enhanced enzymes (e.g. cellulases), substrates or enhancers, and more importantly, to design and synthesize novel classes of compounds with modified or enhanced EXPB1 activity.

(ii) In Silico Analysis and Design

Although the invention will facilitate the determination of actual crystal structures comprising an EXPB1 and a compound, which interacts with the EXPB1, current computational techniques provide a powerful alternative to the need to generate such crystals and generate and analyze diffraction date. Accordingly, a particularly preferred aspect of the invention relates to in silico methods directed to the analysis and development of compounds which interact with EXPB1 structures of the present invention.

Determination of the three-dimensional structure of EXPB1 provides important information about the binding sites of EXPB1, particularly when comparisons are made with similar expansins, and grass pollen allergens. This information may then be used for rational design and modification of EXPB1 substrates and inhibitors, e.g. by computational techniques which identify possible binding ligands for the binding sites, by enabling linked-fragment approaches to drug design, and by enabling the identification and location of bound ligands (e.g. including those ligands mentioned herein above) using X-ray crystallographic analysis. These techniques are discussed in more detail below.

Thus as a result of the determination of the EXPB1 three-dimensional structure, more purely computational techniques for chemical compound design may also be used to design structures whose interaction with EXPB1 is better understood (for an overview of these techniques see e.g. Walters et al (Drug Discovery Today, Vol. 3, No. 4, (1998), 160-178; Abagyan, R.; Totrov, M. Curr. Opin. Chem. Biol. 2001, 5:375-382). For example, automated ligand-receptor docking programs (discussed e.g. by Jones et al. in Current Opinion in Biotechnology, Vol. 6, (1995), 652-656 and Halperin, I.; Ma, B.; Wolfson, H.; Nussinov, R. Proteins 2002, 47:409-443), which require accurate information on the atomic coordinates of target receptors may be used.

The aspects of the invention described herein which utilize the EXPB1 structure in silico may be equally applied to both the EXPB1 structure of disclosed herein and the models of target EXPB1 proteins obtained by other aspects of the invention. Thus having determined a conformation of an EXPB1 by the method described above, such a conformation may be used in a computer-based method of rational drug design as described herein. In addition the availability of the structure of the EXPB1 will allow the generation of highly predictive models for virtual library screening or compound design.

Accordingly, the invention provides a computer-based method for the analysis of the interaction of a molecular structure with an EXPB1 structure of the invention, which comprises: providing the structure of an EXPB1 of the invention; providing a molecular structure to be fitted to said EXPB1 structure; and fitting the molecular structure to the EXPB1 structure.

In an alternative aspect, the method of the invention may utilize the coordinates of atoms of interest of the EXPB1 binding region, which are in the vicinity of a putative molecular structure, for example within 10-25 Å of the catalytic regions or within 5-10 Å of a compound bound, in order to model the pocket in which the structure binds. These coordinates may be used to define a space, which is then analyzed “in silico”. Thus the invention provides a computer-based method for the analysis of molecular structures which comprises; providing the coordinates of at least two atoms of an EXPB1 structure of the invention (“selected coordinates”); providing the structure of a molecular structure to be fitted to said coordinates; and fitting the structure to the selected coordinates of the EXPB1.

In practice, it will be desirable to model a sufficient number of atoms of the EXPB1 as defined herein, which represent a binding groove, e.g. the atoms of the residues identified in residues G1229-N157 which also preferably maintains the binding motifs TWYG, GGACG, HFD. Thus, in this embodiment of the invention, there will preferably be provided the coordinates of at least 5, preferably at least 10, more preferably at least 50 and even more preferably at least 100, e.g. at least 500 such as at least 1000, selected atoms of the EXPB1 structure.

Although every different compound metabolized by EXPB1 may interact with different parts of the binding pocket of the protein, the structure of this EXPB1 allows the identification of a number of particular sites which are likely to be involved in many of the interactions of EXPB1 with a candidate compound. The residues are set out in FIGS. 2 and 3. Thus in this aspect of the invention, the selected coordinates may comprise coordinates of some or all of these residues.

In order to provide a three-dimensional structure of compounds to be fitted to an EXPB1 structure of the invention, the compound structure may be modeled in three dimensions using commercially available software for this purpose or, if its crystal structure is available, the coordinates of the structure may be used to provide a representation of the compound for fitting to an EXPB1 structure of the invention.

The binding pockets of cytochrome EXPB1 molecules are of a size which can accommodate more than one ligand. Indeed, some interactions may occur as a result of interaction of the compounds within the binding pocket of the same EXPB1. In any event, the findings of the present invention may be used to examine or predict the interaction of two or more separate molecular structures within the EXPB1 binding pocket of the invention.

Thus the invention provides a computer-based method for the analysis of the interaction of two molecular structures within an EXPB1 binding pocket structure, which comprises: providing the EXPB1 structure; providing a first molecular structure; fitting the first molecular structure to said EXPB1 structure; providing a second molecular structure; and fitting the second molecular structure to a different part said EXPB1 structure.

Optionally the method of analysis further comprises providing a third molecular structure and also fitting that structure to the EXPB1 structure. Indeed, further molecular structures may be provided and fitted in the same way.

In one aspect, one or more of the molecular structures may be fitted to one or more of the polysaccharide binding area, residues G129 through N157 of the EXPB1 binding groove mentioned above, and one or more of the other molecular structures may be fitted to coordinates of amino acids from another part of the EXPB1 binding pocket, such as another part of the ligand-binding region.

Following the fitting of the molecular structures, a person of skill in the art may seek to use molecular modeling to determine to what extent the structures interact with each other (e.g. by hydrogen bonding, other non-covalent interactions, or by reaction to provide a covalent bond between parts of the structures) or the interaction of one structure with EXPB1 is altered by the presence of another structure.

The person of skill in the art may use in silico modeling methods to alter one or more of the structures in order to design new structures which interact in different ways with EXPB1, so as to speed up or slow down their metabolism, as the case may be.

Newly designed structures may be synthesized and their interaction with EXPB1 may be determined or predicted as to how the newly designed structure is metabolized by said EXPB1 structure. This process may be iterated so as to further alter the interaction between it and the EXPB1.

By “fitting”, it is meant determining by automatic, or semi-automatic means, interactions between at least one atom of a molecular structure and at least one atom of an EXPB1 structure of the invention, and calculating the extent to which such an interaction is stable. Interactions include attraction and repulsion, brought about by charge, steric considerations and the like. Various computer-based methods for fitting are described further herein.

More specifically, the interaction of a compound or compounds with EXPB1 can be examined through the use of computer modeling using a docking program such as GOLD (Jones et al., J. Mol. Biol., 245, 43-53 (1995), Jones et al., J. Mol. Biol., 267:727-748 (1997)), GRAMM (Vakser, I. A., Proteins, Suppl., 1:226-230 (1997)), DOCK (Kuntz et al, J. Mol. Biol. 1982, 161:269-288, Makino et al, J. Comput. Chem. 1997, 18:1812-1825), AUTODOCK (Goodsell et al, Proteins 1990, 8:195-202, Morris et al, J. Comput. Chem. 1998, 19:1639-1662.), FlexX, (Rarey et al, J. Mol. Biol. 1996, 261:470-489) or ICM (Abagyan et al, J. Comput. Chem. 1994, 15:488-506). This procedure can include computer fitting of compounds to EXPB1 to ascertain how well the shape and the chemical structure of the compound will bind to the EXPB1.

Also computer-assisted, manual examination of the active site structure of EXPB1 may be performed. The use of programs such as GRID (Goodford, J. Med. Chem., 28, (1985), 849-857)—a program that determines probable interaction sites between molecules with various functional groups and an the polysaccharide binding surface—may also be used to analyze the active site to predict, for example, the types of modifications which will alter the rate of conformational change, or cell wall extension a compound or plant cell type.

Computer programs can be employed to estimate the attraction, repulsion, and steric hindrance of the two binding partners (i.e. the EXPB1 and a compound).

If more than one EXPB1 active site is characterized and a plurality of respective smaller compounds are designed or selected, a compound may be formed by linking the respective small compounds into a larger compound, which maintains the relative positions and orientations of the respective compounds at the active sites. The larger compound may be formed as a real molecule or by computer modeling.

Detailed structural information can then be obtained about the binding of the compound to EXPB1, and in the light of this information adjustments can be made to the structure or functionality of the compound, e.g. to alter its interaction with EXPB1. The above steps may be repeated and re-repeated as necessary.

As indicated above, molecular structures, which may be fitted to the EXPB1 structure of the invention, include compounds under development as potential enzymatic agents. The agents may be fitted in order to determine how the action of EXPB1 modifies the agent and to provide a basis for modeling candidate agents, which are metabolized at a different rate by an EXPB1.

Molecular structures, which may be used in the present invention, will usually be compounds under development for pharmaceutical use. Generally such compounds will be organic molecules, which are typically from about 100 to 2000 Da, more preferably from about 100 to 1000 Da in molecular weight. Such compounds include peptides and derivatives thereof, and the like. In principle, any compound under development in the field of enzymology can be used in the present invention in order to facilitate its development or to allow further design to improve its properties.

(iii) Analysis of Compounds in Binding Pocket Regions

Our finding of a long grooved binding region allows the analysis and design methods described in the preceding subsections to be focused on compounds which interact with one or more of the residues which make up this area.

Thus in one embodiment, the present invention provides a method for modifying the structure of a compound (polysaccharide) in order to alter its binding to EXPB1 or hydrolysis when bound to EXPB1, which method comprises: fitting a starting compound to one or more coordinates of at least one amino acid residue of the ligand-binding region of the EXPB1; modifying the starting compound structure so as to increase or decrease its interaction with the ligand-binding region.

In another embodiment, the present invention provides a method for modifying the structure of a compound in order to alter its metabolism by an EXPB1, which method comprises: fitting a starting compound to one or more coordinates of at least one amino acid residue of the ligand-binding region of the EXPB1; modifying the starting compound structure so as to increase or decrease its interaction with the ligand-binding region; wherein said ligand-binding region is defined as including at least one, such as at least two, for example such as at least five, preferably at least ten of the EXPB1 residues in the binding groove.

In another embodiment, the invention provides a method for modifying the structure of a compound in order to alter its binding properties to EXPB1 or cell wall extension when bound, which method comprises: fitting a starting compound to one or more coordinates of at least one amino acid residue of the binding region of the EXPB1; modifying the starting compound structure so as to increase or decrease its interaction with the binding region.

Desirably, in the above aspects of the invention, coordinates from at least two, preferably at least five, and more preferably at least ten amino acid residues of the EXPB1 will be used.

For the avoidance of doubt, the term “modifying” is used as defined in the preceding subsection, and once such a compound has been developed it may be synthesized and tested also as described above.

(viii) Compounds of the Invention.

Where a potential modified compound has been developed by fitting a starting compound to the EXPB1 structure of the invention and predicting from this a modified compound with an altered rate of metabolism (including a slower, faster or zero rate), the invention further includes the step of synthesizing the modified compound and testing it in an in vivo or in vitro biological system in order to determine its activity and/or the rate at which it is metabolized.

The method comprises: (a) providing EXPB1 under conditions where, in the absence of modulator, the EXPB1 is able to metabolize known substrates; (b) providing the compound; and (c) determining the extent to which the compound is metabolized in the presence of EXPB1 or (d) determining the extent to which the compound inhibits metabolism of a known substrate of EXPB1.

More preferably, in the latter steps the compound is contacted with EXPB1 under conditions to determine its function.

For example, in the contacting step above the compound is contacted with EXPB1 in the presence of the compound, and typically a buffer and substrate, to determine the ability of said compound to inhibit EXPB1 or to be metabolized by EXPB1. So, for example, an assay mixture for EXPB1 may be produced which comprises the compound, substrate and buffer.

In another aspect, the invention includes a compound, which is identified by the methods of the invention described above.

Following identification of such a compound, it may be manufactured and/or used in the preparation, i.e. manufacture or formulation, of a composition such as an enzymatic composition used in ethanol production, paper recycling or other plant cell extension industrial applications.

Thus, the present invention extends in various aspects not only to a compound as provided by the invention, but also to formulations including acceptable excipients, vehicles or carriers, and optionally other ingredients.

The above-described processes of the invention may be iterated in that the modified compound may itself be the basis for further compound design.

By “optimizing the structure” we mean e.g. adding molecular scaffolding, adding or varying functional groups, or connecting the molecule with other molecules (e.g. using a fragment linking approach) such that the chemical structure of the modulator molecule is changed while its original modulating functionality is maintained or enhanced. Such optimization is regularly undertaken during chemical compound development programs to e.g. enhance potency, promote pharmacological acceptability, increase chemical stability etc. of lead compounds.

Modification will be those conventional in the art known to the skilled medicinal chemist, and will include, for example, substitutions or removal of groups containing residues which interact with the amino acid side chain groups of an EXPB1 structure of the invention. For example, the replacements may include the addition or removal of groups in order to decrease or increase the charge of a group in a test compound, the replacement of a charge group with a group of the opposite charge, or the replacement of a hydrophobic group with a hydrophilic group or vice versa. It will be understood that these are only examples of the type of substitutions considered by medicinal chemists in the development of new pharmaceutical compounds and other modifications may be made, depending upon the nature of the starting compound and its activity.

EXAMPLE 1
Crystal Structure and Activities of EXPB1 (Zea m 1), a Beta-Expansin and Group-1 Pollen Allergen from Maize

Expansins are small extracellular proteins that promote turgor-driven extension of plant cell walls. EXPB1 (also called Zea m 1) is a member of the β-expansin subfamily known in the allergen literature as group-1 grass pollen allergens. EXPB1 induces extension and stress relaxation of grass cell walls. To help elucidate expansin's mechanism of wall loosening, we determined the structure of EXPB1 by X-ray crystallography to 2.75 Å resolution. EXPB1 consists of two domains closely packed and aligned so as to form a long, shallow groove with potential to bind a glycan backbone of ˜10 sugar residues.

The structure of EXPB1 domain 1 resembles that of family-45 glucoside hydrolase (GH45), with conservation of most of the residues in the catalytic site. However, EXPB1 lacks a second aspartate that serves as the catalytic base required for hydrolytic activity in GH45 enzymes. Domain 2 of EXPB1 is an immunoglobulin-like β-sandwich with aromatic and polar residues that form a potential surface for polysaccharide binding in line with the glycan binding cleft of domain 1. EXPB1 binds to maize cell walls, most strongly to xylans, causing swelling of the cell wall. Tests for hydrolytic activity by EXPB1 with various wall polysaccharides proved negative. Moreover, GH45 enzymes and a GH45-related protein called “swollenin”, lacked wall extension activity comparable to that of expansins. We propose a model of expansin action in which EXPB1 facilitates the local movement and stress relaxation of arabinoxylan-cellulose networks within the wall by noncovalent rearrangement of its target.

Prior to maturation plant cells typically experience a period of prolonged cell enlargement, often resulting in a >10³fold increase in volume. The impressive height of trees, some exceeding 100 m, depends on such enlargement, which entails massive vacuolar expansion and irreversible yielding of the cellulosic cell wall. In physical terms, the rate-limiting process for cell enlargement resides within the cell wall, which must be loosened so as to allow wall stress relaxation and consequent water uptake for vacuole enlargement and stretching of the wall (1, 2). Currently, the only plant proteins shown to cause cell wall relaxation are expansins (3, 4), although xyloglucan endotransglucosylase, pectate lyase, cellulase and other enzymes participate in cell wall restructuring during cell growth (5-8).

Expansins were originally discovered in a “fishing expedition” for catalysts of cell wall extension (9, 10). When walls are clamped in tension and incubated in acidic buffer, these proteins rapid induce wall extension and enhance wall stress relaxation. Their biological role in promoting cell enlargement is amply supported by in-vitro and in-vivo experiments, as well as by studies of gene expression, gene silencing, and ectopic expression (3, 11-13). In addition to cell enlargement, expansins are also implicated in other developmental processes where wall loosening occurs, such as in fruit softening, organ abscission, seed germination, and pollen tube invasion of the grass stigma (14-17).

Two expansin families with wall-loosening activity have been identified, named α-expansins (EXPA) and β-expansins (EXPB); both are found in all groups of land plants, from mosses to flowering plants (3, 18). Although they have only ˜20% amino acid identity, EXPA and EXPB proteins are of similar size (˜27 kD), their sequences align well with one another and they contain a number of conserved residues and characteristic motifs distributed throughout the length of the protein. EXPA and EXPB appear to act on different cell wall components, but their native targets have not yet been well defined.

A subset of β-expansins is known in the immunological literature as group-1 grass pollen allergens (19-21). These β-expansins are abundantly and specifically expressed in grass pollen, causing hay fever and seasonal asthma in an estimated 200-400 million humans (22, 23). The extraordinary abundance of group-1 allergens—comprising up to 4% of the protein extracted from grass pollen (24)—is unique (as far as we know) in the world of expansins, which are typically found in very low abundance and tightly bound to the cell wall. The abundance of group-1 allergens in grass pollen bespeaks a unique biological role, namely to loosen the cell walls of the grass stigma and style, thereby aiding pollen tube penetration and assisting delivery of its two sperm cells to the ovule, where a double fertilization occurs, forming the diploid zygote and the triploid endosperm. Seed development follows, and because cereal grasses provide the largest food source for humanity (e.g. rice, maize, wheat, and barley, to name but a few), the importance of these events for human welfare is hard to overestimate.

Other genes in the β-expansin family are expressed in a variety of other tissues in the plant body and in general lack the specific allergenic epitopes characteristic of group-1 allergens (24, 25). These so-called “vegetative β-expansins” are thought to have cell wall loosening activity and substrate specificity similar to the group-1 allergens, but these inferences have yet to be demonstrated experimentally.

The mechanism by which expansins loosen cell walls has not yet been worked out in molecular detail. Plant cell walls consist of a scaffold of long cellulose microfibrils ˜4 nm in diameter, embedded in a matrix of cellulose-binding glycans, such as xyloglucan and arabinoxylan, and gel-forming pectic polysaccharides (FIG. 1). The cellulose-binding glycans form a stable network with the cellulose microfibrils by binding to their surface via hydrogen bonds between hydroxyl groups and via van der Waals forces between the sugar rings; the network is further stabilized by calcium ions and borate diesters that link pectic polysaccharides together. Cell walls also contain small amounts of structural proteins with a reinforcing role (26, 27). Wall expansion entails rearrangement or modification of the matrix to allow turgor-driven movement or slippage of cellulose microfibrils within the matrix (1).

Most of the biochemical work on expansins to date has focused on α-expansins, which do not hydrolyze the major structural polysaccharides of the wall and indeed are devoid of every enzyme activity assayed to date (28). Our current model proposes that α-expansins disrupt the polysaccharide complexes that link cellulose microfibrils together. The pollen β-expansins (group-1 allergens) have a marked loosening action on cell walls from grasses, but not from dicots, whereas the reverse is true for α-expansins; therefore it seems that the two forms of expansin target different components of the cell wall (21, 24). Grass cell walls are notable for containing relatively small amounts of xyloglucan and pectin, which are replaced with β-(1→3),(1→4)-D-glucan and glucuronoarabinoxylan (29)—two potential targets of β-expansins in their wall-loosening activity.

Sequence analysis suggests that expansins consist of two domains (2, 3). The putative N-terminal domain (D1) has distant sequence similarity (˜20% identity) to the catalytic domain of family-45 glycoside hydrolases (GH45; http://afmb.cnrs-mrs.fr/CAZY/). Despite this resemblance, α-expansins do not hydrolyze wall polysaccharides and so the sequence similarity is enigmatic. The C-terminal domain (D2) has sequence similarity (from 35% to <10% identity) to another class of allergens, the group-2/3 grass pollen allergens, whose biological function is unknown (30).

In this study we present the crystal structure of a native β-expansin purified from maize pollen. In the allergen field it is designated Zea m 1 isoform d, whereas by expansin nomenclature it is called EXPB1 (GenBank accession AAO45608). The allergen name “Zea m 1” encompasses a group of at least four pollen proteins (EXPB1, EXPB9, EXPB10, EXPB11) in two rather divergent sequence classes (24). EXPB1 is the most abundant of the maize group-1 allergens. We also test EXPB1 for binding and activity on cell walls. At the end we discuss a molecular model of expansin action that is consistent with its structure and known biophysical and biochemical activities.

Results

EXPB1 has two closely-packed domains. Native EXPB1 was purified from maize pollen and crystallized in 15% (w/v) polyethylene glycol 4000 with 0.1 or 0.2 M ammonium sulfate. Two crystals were analyzed, yielding X-ray diffraction patterns consistent with the monoclinic C₂space group. EXPB1 structure was solved and refined to 2.75 Å resolution (see Methods) with a crystallographic R-factor of 0.233 and an R-free of 0.291 (Table 1).

EXPB1 contains two domains (residues 19-140 [D1] and 147-245 [D2]) connected by a short linker (residues 141-146) and aligned end to end so as to make a closely-packed irregular cylinder ˜66 Å long and 26 Å in diameter (FIG. 2A). At its N-terminus EXPB1 has a flexible sequence (residues 1-18) containing hydroxyproline (O9) and a glycan attached to N10, part of the glycosylation consensus sequence NXT. The end of the glycan comes close to the polysaccharide-binding groove (see figures) of the symmetry-related protein in the crystalline lattice, with one of the mannose residues stacking against the planar surface formed by residues Gly39 and Gly40 and stabilized further by two hydrogen bonds with the side chain of D37. These interactions with the symmetry-related protein account in part for the unusual ordering of the glycan, as well as the ability to crystallize the glycosylated protein.

Based on its electron density, our model of this N-linked glycan consists of a β-(1→4)-linked backbone of GlcNac₁GlcNac₂Man₃with two Man residues and a Xyl residue attached to Man₃and a Fuc residue linked to GlcNac₁. Such so-called paucimannosidic-type N-linked glycans are characteristically processed in the Golgi and in post-Golgi steps (31).

Previous analysis (2, 3) indicated that D1 has distant sequence similarity to members of glycoside hydrolase family 45 (GH45), whose members have been characterized as inverting endo-β-(1→4)-D-glucanases (2, 3, 32, 33). Superposition of D1 with a GH45 enzyme (PDB #4ENG) using the secondary structure matching algorithm in CCP4 (34) gives good overlap of the two structures for 84 residues (60%) of the peptide backbone of D1 (FIG. 2B), with an root mean square deviation (rmsd) of 2.5 Å. Two of the three disulfide bonds in D1 superimpose exactly with 4ENG disulfides (the exception being C78-C84). Likewise, all of the β-strands in D1 superimpose on β-strands of 4ENG, although the β-strands in EXPB1 are generally shorter. Both structures have short α-helices, but these do not overlap in the two structures.

What is missing in EXPB1 is a residue corresponding to D10*, the catalytic base required for glucan hydrolysis by GH45 enzymes (35). As indicated in FIG. 2C, D10* is located on a loop that is not aligned with any part of EXPB1. EXPB proteins do have a conserved acidic residue, D37, which is located in a loop (residues 29-38) in the general vicinity corresponding to D10* in 4ENG. This loop is well resolved in D1. However, D37 is located too far from D107 and Y27 to function as the required base. In 4ENG, the catalytic carboxylate groups are located 8.5 Å apart, which is sufficient distance to accommodate a water molecule needed for hydrolysis (35). In D1, the carboxylates for D107 and D37 are 15 Å apart, too distant for this catalytic mechanism. Moreover, simple lateral movement of the loop to bring D37 into a correct position seems unlikely as the loop residues following D37 are rigidly held in place by a several stabilizing interactions. Thus, a key part of the catalytic machinery required for hydrolytic activity of GH45 enzymes is lacking in EXPB1.

We conclude that, despite the structural similarity of D1 to GH45, EXPB1 does not induce wall extension via wall polysaccharide hydrolysis.

Structure of Domain 2 (D2). Residues 147-245 of EXPB1 make up a second domain (D2) composed of eight β strands assembled into two antiparallel β sheets (FIG. 2A. The two β sheets are at slight angles to each other and form a β-sandwich similar to the immunoglobulin fold. D2 has 36% sequence identity with Phl p 2, a group-2/3 grass pollen allergen (PDB #1WHO), and superposition of the two structures shows them to have identical folds (rmsd of 1.3 Å; FIG. 2D). In comparing the two structures, we find that D2 tends to have shorter β strands compared with Phl p 2 and the two proteins deviate slightly in the loop regions connecting the β-strands.

D1 and D2 form a long potential polysaccharide-binding site. The two EXPB1 domains align so as to form a long, shallow groove with highly conserved polar and aromatic residues suitably positioned to bind a twisted polysaccharide chain of 10 xylose residues (FIGS. 2E-G). The groove extends from the conserved G129 at one end of D1, spans across a stretch of conserved residues in D1 and D2 (see numbered residues in FIG. 2E as well as annotated sequence logo in FIG. 3) and ends at N157, a distance of some 47 Å. Many of the conserved residues common to EXPA and EXPB make up this potential binding surface, including residues in the classic expansin motifs TWYG, GGACG, and HFD (see FIG. 3).

The openness of the long groove may enable EXPB1 to bind polysaccharides that are part of a bulky cell wall complex, such as on the surface of cellulose; that openness may also be important for binding branched glycans such as arabinoxylan which itself binds to the surface of cellulose microfibrils. Because EXPB1 binds preferentially to xylans (see below), we have modeled an arabinoxylan, characteristic of grass cell walls, bound to the long groove of EXPB1 (FIG. 2G). From this model it is clear that the open groove of EXPB1 can accommodate the side chains found on such polysaccharides.

A second conserved surface in D2 is far removed from D1 (arrows in FIG. 2G). There is a shallow cup formed by the conserved W232 and F210. Adjacent to this pocket is a hydrophobic surface patch formed by the conserved residues P209, P229, V227 and Y238. The pocket and adjacent region could provide a second glucan binding surface for ˜3 residues.

Binding. EXPB1 bound to isolated maize cell wall (FIG. 4B). We observed that cell walls incubated with EXPB1 swelled significantly when compared with control cell walls (FIG. 4D). When purified polysaccharide fractions were immobilized onto nitrocellulose membranes, EXPB1 bound preferentially to xylans, with negligible binding to β-(1→3),(1→4) D-glucan and glucomannan (FIG. 4C). Intermediate binding to xyloglucan was observed. Specific binding to cellulose and to nitrocellulose was also seen, although with less avidity than to xylan (A. Tabuchi and D. J. Cosgrove, manuscript in preparation).

Discussion

With the molecular structure of EXPB1 in hand, we can examine previous inferences about expansin structure and its mechanism of cell wall loosening, but first the use of the group-1 pollen allergen for this study merits comment. Unlike other forms of expansin, which are found in very low abundance and have low solubility, the group-1 allergens are produced in copious amounts by grass pollen, from which they are readily extracted, purified, and concentrated to high levels without precipitation. Moreover, grasses produce abundant pollen, with maize being an especially liberal donor. In contrast to recombinant forms, use of the native protein insures correct processing and post-translational modifications. We note that expression of active expansins in various recombinant systems has proved problematic, due to improper folding, aggregation and hyperglycosylation (M. Shieh and D. J. Cosgrove, unpublished data). Other forms of {tilde over (□)}expansin (e.g. the vegetative homologs) require harsh conditions to extract them from plant tissues (38), resulting in denatured protein; in soybean cultures an EXPB accumulates in the medium, but in a degraded and inactive form (39). EXPA proteins have been purified from various plant tissues, but in our experience they are difficult to concentrate to levels suitable for crystallization.

The high solubility and abundance of the group-1 allergens thus commends them for crystallization studies, but it should be noted that some of their biochemical properties may be specialized for their unique biological role in grass pollination. A case in point is their atypical pH dependence (maximum activity at pH 5.5; (24)), which is shifted to less acidic values than that found for other expansins. Likewise, their high solubility seems to be exceptional. Nevertheless, the general features of EXPB1 structure should prove to be common to the whole expansin family.

EXPB1 is composed of two domains. Although D1 structurally resembles GH45 and indeed has conserved much of the GH45 catalytic site, it lacks the second Asp residue—the catalytic base—required for hydrolytic activity in GH45 enzymes (33, 35). Thus, expansin's lack of wall polysaccharide hydrolytic activity, documented here for EXPB1 and in previous work for EXPA (28, 40), can be understood in structural terms as due to the lack of the required catalytic base. Furthermore, our finding that bona fide GH45 enzymes lack expansin's wall extension activity lends additional support to the conclusion that expansin does not loosen the cell wall by polysaccharide hydrolysis.

D2 as binding module? We previously speculated that D2 may be a carbohydrate-binding module (CBM) (2, 4). This notion gains indirect support from the structure of D2, in which two surface aromatic residues (W194, Y160) are in line with two aromatic residues (W26, Y27) in D1, forming part of an extended, open, and highly conserved surface in EXPB1. D2 has an immunoglobulin-like fold. Proteins with this fold form a large superfamily of β-sandwich proteins implicated in binding interactions, but lacking in enzymatic activity (41). At least 16 of the currently recognized CBM families in the Carbohydrate-Active Enzymes (CAZY) database (http://afmb.cnrs-mrs.fr/CAZY/) have a β-sandwich fold. However, the specific fold topology of D2 does not match any of these CBM folds and D2 lacks a bound metal atom, found in nearly all of the β-sandwich CBMs (42).

Nevertheless, from the structure of EXPB1 we expect that D2 aids glycan binding, particularly via the two surface aromatic residues W194 and Y160, aided by polar residues S193, R199, C156 and N157. These potential sugar-binding residues do not correspond to those inferred from a homology model of Lol p 1, a group-1 allergen from rye grass (43). In this model, which was based on the structure of Phl p 2, a group-2 allergen (30, 44), the authors identified two potential polysaccharide binding surfaces, one of which corresponds to the buried D2 face contacting D1.

It is notable that endoglucanases are most often found in nature as modular enzymes, coupled to a CBM via a long, highly glycosylated linker. Crystallization of intact GH45 enzymes with their CBMs has not yet been achieved, probably because the two domains do not maintain a fixed spatial relationship to each other. This difficulty of crystallization is a common experience with many CBM-coupled enzymes, and so successful crystallization of the two-domain EXPB1 is notable in this regard. In EXPB1 the linker is very short and the multiple contacts between D1 and D2 enable close coupling of the two domains, which may function as a single unit in binding the cell wall.

Expansins as cysteine proteases? A controversial hypothesis has been proposed that group-1 allergens are papain-related cysteine proteinases, with conservation of papain's active site residues C25, H159 and N175 (the “catalytic triad”) (45, 46). According to this hypothesis, C73 in EXPB1 should correspond to papain's C25. However, from the structure of EXPB1 we see that C73 participates in a disulfide bond conserved with GH45 enzymes, is relatively inaccessible, and is nowhere near the conserved surface. Moreover, the residues claimed to correspond to papain's H159 and N175 are dispersed in D2, remote from C73 and are not conserved in expansins. We conclude that the resemblance to papain suggested by Grobe et al. (45, 46) is not supported by our crystallographic model of EXPB1.

The conserved surface of EXPB1 does contain two Cys residues (C58, C156), but their environment does not resemble that of papain's active site. C58, which is conserved in about half of the EXPB family, is relatively inaccessible, being mostly buried underneath Y27 at the bottom of the extended groove. C156 not conserved in the EXPB family, but is usually replaced by serine. Experimental assays failed to detect proteinase activity in native EXPB1 (47). Moreover, the group-1 allergens are noted for their remarkable stability, which is also the case for EXPB1. We deem it likely that recombinant expression of EXPB in Pichia induced a host protease that accounted for the protein instability observed by Grobe et al. (45, 46). In fact, such host proteinase induction has been reported upon recombinant expression of a group-1 allergen (48).

Comparison with vegetative β-expansins and with α-expansins. EXPB1 is a member of the group-1 grass pollen allergens, which comprise a subset of the larger EXPB family. The EXPB family is notably larger in grasses than in other groups of land plants, and part of this expansion involved the unique evolution and radiation of the pollen allergen class of EXPBs, which are encoded by multiple genes (49). For instance, we classified 5 of the 19 EXPB genes in the rice genome as group-1 allergens (49). Multiple EXPB genes of the pollen allergen class may account in part for the numerous group-1 “isoallergens” found in grass pollen (19, 20, 50, 51).

There are minor conserved differences between the allergen class and the remaining “vegetative” EXPBs. These are so slight that we expect the structural features of EXPB1 are characteristic of the vegetative EXPBs, with one exception: the N-terminal extension in EXPB1 contains a motif (VPPGPNITT) that is consistently found, with only minor variation, in group-1 grass pollen allergens, but not in other EXPBs. This motif contains one or more hydroxyprolines and a glycosylated asparagine, features common to the pollen allergen class of EXPB (52). The function of this N-terminal extension is unknown, but it may play a role in protein recognition, transport, packaging and processing by the pollen secretory apparatus. Additionally, the glycosylated extension may contribute to the exceptional solubility of the group-1 allergens (other expansins characterized to date have very low solubility) or may interact with other components of the cell wall. While this motif is a unique hallmark of the group-1 allergens, many EXPB proteins lack an N-terminal extension altogether, and so it is not an essential part of expansin function. However, an N-terminal extension with similar post-translational modifications was found as part of an EXPB expressed in soybean cell cultures (39).

The good sequence alignment and conservation of motifs between the EXPB and EXPA families make it likely that EXPA proteins will have the same three-dimensional structure as reported here for EXPB1. There are two notable regions where EXPA and EXPB differ. EXPA has an additional stretch of ˜12 amino acids in the region corresponding to E99/P100 in EXPB1. E99 and P100 are part of a loop between β strands IV and V in D1; these residues form part of the upraised flank to the left of the long groove identified in FIG. 2. The additional residues in EXPA may form a larger shoulder flanking this groove, stabilized by a disulfide bond between a pair of cysteines in this loop that are conserved in the EXPA family but are lacking in many EXPBs, mostly notably absent in the pollen allergens. This idea gains support from the structure of another GH45 enzyme (PDB #1WC2 (53)) which contains just such a loop (residues 102-114) stabilized by a disulfide bond. The loop creates a shoulder abutting the catalytic cleft. EXPAs therefore may have a steeper binding cleft than that does EXPB1.

A second difference is that EXPAs lack a segment corresponding to G120-H127 in EXPB1. This segment, which contains few conserved residues, forms α-helix c and constitutes part of the surface of the pointed end of D1. This surface is remote from the conserved regions we have identified, and so is unlikely to affect activity.

Allergenic epitopes. Allergies to grass pollen are widespread, afflicting an estimated 200-400 million people, and numerous studies have concluded that the group-1 allergens are the most important allergenic components of grass pollen (23, 23, 54, 55). Maize EXPB1 and its orthologs in turf grasses share common epitopes, as judged by antibody cross reactivity, with the predominant epitopes found in the protein portion of the molecule and the glycosyl residues being of secondary antigenic significance (52, 56, 57). The dominant group-1 allergenic epitopes, which have been identified by epitope mapping studies, can be readily located on the surface of EXPB1. For instance, the 15-residue c98 epitope identified by Ball et al. (58) includes D107 in the conserved catalytic site of EXPB1, but also includes residues that are exposed on the opposite side of the protein. “Site D” identified by Hiller et al. (59) overlaps part of the extended conserved groove of D1 containing the motif TWYG28 (FIG. 2E), whereas “site A” identified by Esch and Klapper (60) includes the small conserved pocket containing W232 and Y238, found on the far side of D2, as indicated in FIGS. 2E, G. This pocket is also part of “peptide 5” (22), a synthetic peptide derived from B cell epitopes of Phl p 1, the group-1 allergen of timothy grass pollen. Antibodies against peptide 5 showed great potency in reducing binding of IgEs from patients with strong grass pollen allergens, and so this peptide was considered a potentially useful component of an epitope-based vaccine for treating patients with severe allergies to grass pollen (22). With the structure of EXPB1 in hand, one may consider designing synthetic peptides that more closely resemble the natural epitopes occurring on the conserved surface of group-1 allergens. These may be of use for immunotherapy as well as mechanistic studies concerning the molecular and cellular bases for the potency of these proteins as allergens.

In view of the sequence conservation within the EXPB family, as well as within the entire expansin superfamily, it is surprising that the dominant antigenic epitopes of the group-1 allergens are not shared by vegetative EXPBs or by EXPA members. Nevertheless, this seems to be the case because antibodies raised against the group-1 allergens do not recognize other forms of expansin. This is indeed fortunate, for otherwise persons with strong allergies to grass pollen would also be allergic to fresh fruits, vegetables, grains and other plant tissues that express members of this large gene family that is ubiquitous in plants.

A molecular model of wall loosening by expansins. Expansin action may be summarized as follows: the protein binds one or more wall polysaccharides and within seconds induces wall stress relaxation followed by wall extension, without hydrolysis of the wall polymers. There is no requirement for ATP or other source of chemical energy, and the wall continues to extend so long as the wall bears sufficient tension and expansin is present (that is, expansin acts catalytically, not stoichiometrically).

In the case of EXPB1, we imagine that stress relaxation begins when it binds a taut arabinoxylan tethered to a cellulose microfibril, causing local release of the arabinoxylan from the cellulose surface. Movement of the β-expansin along the arabinoxylan-cellulose junction would enable it to unzip the hydrogen bonds between the polysaccharides, relaxing the taut tether and allowing turgor-driven displacement of cellulose and arabinoxylan, which may then reassociate in a relaxed state to restore wall strength. During this movement, the two expansin domains might shift in a hinge-like manner, binding and letting go of the arabinoxylan independently of each other, leading to an inchworm-like movement along the polysaccharide. We estimate that as little as 10° shift in angle between domains could cause a one-residue dislocation of the polysaccharide along the binding surface.

To assess the feasibility of such inter-domain movement, we estimated the buried surface area between the two domains, using CCP4. The value is 589 Å, which is indicative of a weak inter-domain interaction (61), is consistent with domain movements as imagined above. A potential source of energy for these movements is the mechanical strain energy stored by the taut polysaccharide in a turgor-stretched cell wall. In this model, expansin acts as molecular device that uses the strain energy stored in a taut cellulose-binding glycan to help dissociate the glycan from the surface of cellulose.

Materials and Methods

Protein Purification, Crystallization and Data Collection. Native Zea m 1 was extracted from pollen of field-grown maize plants at 4° C. in 0.125 M sodium carbonate and then purified to electrophoretic homogeneity in the presence of 5 mM dithiothreitol using two chromatographic steps as described (24). With this method four Zea m 1 isoforms were readily distinguished and we used the most abundant isoform, Zea m 1d (=EXPB1), for crystallization and activity assays. For the binding experiments, EXPB1 was further purified by HPLC on a reverse phase column (Discovery C8, 15 cm×4.6 mm i.d., 5 μm, Supelco) pre-equilibrated with 10% acetonitrile containing 0.1% trifluoroacetic acid. Bound protein was eluted at 1 mL min⁻ with a linear gradient of 22 to 90% acetonitrile in the same solution for 20 min at a flow rate of 1 mL min⁻, at 25° C. We confirmed wall extension activity of EXPB1 purified in this way.

Crystals were grown at 21° C. for 9 days using EXPB1 at 10.5 mg/mL in 100 mM Na acetate, pH 4.6, in 5-μL hanging drops, with addition of 5-μL precipitant (15% (w/v) polyethylene glycol 4000 with 0.1 or 0.2 M ammonium sulfate) and with 1-mL reservoir volume. Two crystals were analyzed, yielding diffraction patterns consistent with the monoclinic C₂space group. Crystal 1 had unit cell dimensions of a=113.7 Å, b=45.2 Å, and c=70.3 Å, with angles α=90.0°, β=124.6°, and γ=90.0°; crystal 2 had unit cell dimensions of a=112.6 Å, b=44.4 Å, and c=69.6 Å, with angles α=90.0°, β=124.4°, and γ=90.0°.

Data were collected using a RIGAKU RU200 rotating anode X-ray generator with CuK□ radiation, operating at 5 KW of power (50 kV, 100 mA) (Molecular Structure Corporation, The Woodlands, Tex.). Three-degree oscillation frames, each exposed for 120 minutes were collected on an R-AXIS IV detector. The two crystals were used to get a 93% complete dataset. DENZO and SCALEPACK software suite (62) were used for data processing.

Structure Solution and Refinement. Our final model of EXPB1 structure was based on the native crystal data set and was solved by molecular replacement calculations using the program AmoRe (63) with the structure of Phl p 1 (PDB entry code 1N10) which has 58% amino acid identity with EXPB1 over 240 residues. EXPB1 has four more residues at its C-terminus. The best molecular replacement solution in AMoRE was obtained by deleting the first 13 residues of the N-terminus (attempts that included this stretch did not yield a solution) and by including all the side-chains for the rest of the protein (attempts with just the backbone atoms did not yield a good solution as well) and including all the available data to 2.75 Å. The correlation co-efficient and the R-factor for the best solution was 55.1 and 51.0 respectively. The next best solution had an inferior correlation co-efficient and R-factor of 49.3 and 53.9, enabling us to proceed with further refinement and model building with confidence. For further refinement details and comparison with the 1N10 structure, see supplemental text, published on the PNAS web site. Coordinates and structure factors of the structure have been deposited in the protein data bank (PDB code 2HCZ; (64)). A summary of the refinement results is given in Table 1 (on PNAS web site).

Polysaccharide Hydrolysis. Two mg of dye-coupled insoluble polysaccharides (AZCL-polysaccharides, Megazyme, Wicklow, Ireland) were suspended in 100/L buffer (50 mM sodium acetate, pH 4.5, with 1 mM NaN₃and 10 mM dithiothreitol) and incubated with shaking at 30° C. for 48 h+/−30 μg of EXPB1. At the end of the incubation, 300 μL of 2.5% Trizma base was added to each tube to stop reaction, the suspension was centrifuged, and the absorbance (590 nm) of the supernatant was measured.

Binding. Cell walls were collected from maize silks, cleaned by phenol/acetic acid/water washes (65) and lyophilized. EXPB1 was purified on a CM-Sepharose Fast Flow (Amersham Biosciences) column in a LP system (Bio-Rad) (24). EXPB1 (10 μg) was incubated with 1 mg cell wall in 400 μL of 50 mM sodium acetate, pH 5.5, for 1 h at 25° C. with agitation. After incubation, protein remaining in the supernatant was analyzed by SDS-PAGE (12% poly acrylamide), stained with SYPRO Ruby protein gel stain (Bio-Rad).

Commercial polysaccharides dissolved in 20 mM sodium acetate, pH 4.5 (200 μg, oat spelts xylan (Sigma), birch wood xylan (Fluka), barley β-glucan (Sigma, G-6513), konjac glucomanna (Megazyme) and tamarind xyloglucan (Megazyme) were applied to nitrocellulose membranes disks (ca. 7 mm diameter, Protran, BA83, pore size; 0.2 μm, Whatman). The disks were dried at 80° C. overnight. The coated disks were incubated with blocking reagent (Roche) dissolved in 0.1 M maleic acid buffer for 1 h at room temperature to reduce nonspecific binding of EXPB1. After the blocking, the disks were washed with 20 mM Na acetate 5 times for 3 min each, then incubated with EXPB1 (20 μg per tube; purified by reverse-phase chromatography; see above) in 400 μL of 20 mM sodium acetate, pH 5.5 at 25° C. for 1. After the incubation, the supernatant (unbound protein) was analyzed by reverse phase chromatography (above). The amount of EXPB1 bound to the coated nitrocellulose membrane disks was calculated from the reduction in the amount of unbound protein, assessed by reverse-phase HPLC of the supernatant.

Acknowledgments. This work was supported by DOE Grant FG02-84ER13179 and NIH Grant 5R01GM60397 to DJC. We thank: Dr. Greg Farber for instimable advice and assistance with growing the EXPB1 crystals; Dr. Javier Sampedro for useful discussions; Daniel M. Durachko, Edward Wagner and Dr. Hemant Yennawar for expert technical assistance; Dr. Colin Mitchison for gift of the swollenin sample; Dr. Inez Munoz for gift of the TrCel45 sample; Dr. Jan-Christer Janson for gift of the MeCel45 sample.

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TABLE 1

HEADER
ALLERGEN 19-JUN-06 2HCZ

TITLE
CRYSTAL STRUCTURE OF EXPB1 (ZEA M 1), A BETA-EXPANSIN AND

TITLE
2 GROUP-1 POLLEN ALLERGEN FROM MAIZE

COMPND
MOL_ID: 1;

COMPND
2 MOLECULE: BETA-EXPANSIN 1A;

COMPND
3 CHAIN: X;

COMPND
4 SYNONYM: POLLEN ALLERGEN ZEA M 1, ZEA M I

SOURCE
MOL_ID: 1;

SOURCE
2 ORGANISM_SCIENTIFIC: ZEA MAYS;

SOURCE
3 ORGANISM_COMMON: MAIZE;

SOURCE
4 TISSUE: POLLEN

KEYWDS
DOMAIN 1 IS A BETA BARREL AND DOMAIN 2 IS A

IMMUNOGLOBULIN

KEYWDS
2 LIKE BETA-SANDWICH

EXPDTA
X-RAY DIFFRACTION

AUTHOR
N. H. YENNAWAR, D. J. COSGROVE

REVDAT
2 24-OCT-06 2HCZ 1 HEADER

REVDAT
1 22-AUG-06 2HCZ 0

JRNL
AUTH N. H. YENNAWAR, L. C. LI, D. M. DUDZINSKI, A. TABUCHI,

JRNL
AUTH 2 D. J. COSGROVE

JRNL
TITL CRYSTAL STRUCTURE AND ACTIVITIES OF EXPB1 (ZEA M

JRNL
TITL 2 1), A BETA-EXPANSIN AND GROUP-1 POLLEN ALLERGEN

JRNL
TITL 3 FROM MAIZE.

JRNL
REF PROC.NATL.ACAD.SCI.USA V. 103 14664 2006

JRNL
REFN ASTM PNASA6 US ISSN 0027-8424

REMARK
1

REMARK
2

REMARK
2
RESOLUTION. 2.75 ANGSTROMS.

REMARK
3

REMARK
3
REFINEMENT.

REMARK
3
PROGRAM: CNS

REMARK
3
AUTHORS: BRUNGER, ADAMS, CLORE, DELANO, GROS, GROSSE-

REMARK
3
: KUNSTLEVE, JIANG, KUSZEWSKI, NILGES, PANNU,

REMARK
3
: READ, RICE, SIMONSON, WARREN

REMARK
3

REMARK
3
REFINEMENT TARGET: ENGH & HUBER

REMARK
3

REMARK
3
DATA USED IN REFINEMENT.

REMARK
3
RESOLUTION RANGE HIGH (ANGSTROMS):
2.75

REMARK
3
RESOLUTION RANGE LOW (ANGSTROMS):
29.63

REMARK
3
DATA CUTOFF (SIGMA(F)):
0.000

REMARK
3
DATA CUTOFF HIGH (ABS(F)):
NULL

REMARK
3
DATA CUTOFF LOW (ABS(F)):
NULL

REMARK
3
COMPLETENESS (WORKING + TEST) (%): 92.1

REMARK
3
NUMBER OF REFLECTIONS:
7007

REMARK
3

REMARK
3
FIT TO DATA USED IN REFINEMENT.

REMARK
3
CROSS-VALIDATION METHOD: NULL

REMARK
3
FREE R VALUE TEST SET SELECTION: RANDOM

REMARK
3
R VALUE (WORKING SET): 0.233

REMARK
3
FREE R VALUE: 0.290

REMARK
3
FREE R VALUE TEST SET SIZE (%): 4.800

REMARK
3
FREE R VALUE TEST SET COUNT: 367

REMARK
3
ESTIMATED ERROR OF FREE R VALUE: NULL

REMARK
3

REMARK
3
FIT N THE HIGHEST RESOLUTION BIN.

REMARK
3
TOTAL NUMBER OF BINS USED:
8

REMARK
3
BIN RESOLUTION RANGE HIGH (A):
2.75

REMARK
3
BIN RESOLUTION RANGE LOW (A):
2.87

REMARK
3
BIN COMPLETENESS (WORKING + TEST) (%): NULL

REMARK
3
REFLECTIONS IN BIN (WORKING SET): 780

REMARK
3
BIN R VALUE (WORKING SET): 0.4200

REMARK
3
BIN FREE R VALUE: 0.5450

REMARK
3
BIN FREE R VALUE TEST SET SIZE (%): NULL

REMARK
3
BIN FREE R VALUE TEST SET COUNT: 41

REMARK
3
ESTIMATED ERROR OF BIN FREE R VALUE: NULL

REMARK
3

REMARK
3
NUMBER OF NON-HYDROGEN ATOMS USED IN REFINEMENT.

REMARK
3
PROTEIN ATOMS:
1872

REMARK
3
NUCLEIC ACID ATOMS:
0

REMARK
3
HETEROGEN ATOMS:
80

REMARK
3
SOLVENT ATOMS:
17

REMARK
3

REMARK
3
B VALUES.

REMARK
3
FROM WILSON PLOT (A**2): NULL

REMARK
3
MEAN B VALUE (OVERALL, A**2): 56.87

REMARK
3
OVERALL ANISOTROPIC B VALUE.

REMARK
3
B11 (A**2): −1.64000

REMARK
3
B22 (A**2): −4.79800

REMARK
3
B33 (A**2): 6.43900

REMARK
3
B12 (A**2): 0.00000

REMARK
3
B13 (A**2): −1.44900

REMARK
3
B23 (A**2): 0.00000

REMARK
3

REMARK
3
ESTIMATED COORDINATE ERROR.

REMARK
3
ESD FROM LUZZATI PLOT (A): NULL

REMARK
3
ESD FROM SIGMAA (A): NULL

REMARK
3
LOW RESOLUTION CUTOFF (A): NULL

REMARK
3

REMARK
3
CROSS-VALIDATED ESTIMATED COORDINATE ERROR.

REMARK
3
ESD FROM C-V LUZZATI PLOT (A): NULL

REMARK
3
ESD FROM C-V SIGMAA (A): NULL

REMARK
3

REMARK
3
RMS DEVIATIONS FROM IDEAL VALUES.

REMARK
3
BOND LENGTHS (A): 0.008

REMARK
3
BOND ANGLES (DEGREES): 1.65

REMARK
3
DIHEDRAL ANGLES (DEGREES): NULL

REMARK
3
IMPROPER ANGLES (DEGREES): NULL

REMARK
3

REMARK
3
ISOTROPIC THERMAL MODEL: NULL

REMARK
3

REMARK
3
ISOTROPIC THERMAL FACTOR RESTRAINTS. RMS SIGMA

REMARK
3
MAIN-CHAIN BOND (A**2): NULL; NULL

REMARK
3
MAIN-CHAIN ANGLE (A**2): NULL; NULL

REMARK
3
SIDE-CHAIN BOND (A**2): NULL; NULL

REMARK
3
SIDE-CHAIN ANGLE (A**2): NULL; NULL

REMARK
3

REMARK
3
BULK SOLVENT MODELING.

REMARK
3
METHOD USED: NULL

REMARK
3
KSOL: NULL

REMARK
3
BSOL: 46.18

REMARK
3

REMARK
3
NCS MODEL: NULL

REMARK
3

REMARK
3
NCS RESTRAINTS. RMS SIGMA/WEIGHT

REMARK
3
GROUP 1 POSITIONAL (A): NULL; NULL

REMARK
3
GROUP 1 B-FACTOR (A**2): NULL; NULL

REMARK
3

REMARK
3
PARAMETER FILE 1: PROTEIN_REP.PARAM

REMARK
3
PARAMETER FILE 2: CARBOHYDRATE.PARAM

REMARK
3
PARAMETER FILE 3: CNS_TOPPAR:WATER_REP.PARAM

REMARK
3
PARAMETER FILE 4: NULL

REMARK
3
TOPOLOGY FILE 1: NULL

REMARK
3
TOPOLOGY FILE 2: NULL

REMARK
3
TOPOLOGY FILE 3: NULL

REMARK
3
TOPOLOGY FILE 4: NULL

REMARK
3

REMARK
3
OTHER REFINEMENT REMARKS: NULL

REMARK
4

REMARK
4
2HCZ COMPLIES WITH FORMAT V. 3.0, 1-DEC-2006

REMARK
4

REMARK
4
THIS IS THE REMEDIATED VERSION OF THIS PDB ENTRY.

REMARK
4
REMEDIATED DATA FILE REVISION 3.101 (2007-05-29)

REMARK
100

REMARK
100
THIS ENTRY HAS BEEN PROCESSED BY RCSB.

REMARK
100
THE RCSB ID CODE IS RCSB038210.

REMARK
200

REMARK
200
EXPERIMENTAL DETAILS

REMARK
200
EXPERIMENT TYPE: X-RAY DIFFRACTION

REMARK
200
DATE OF DATA COLLECTION: 01-OCT-1998

REMARK
200
TEMPERATURE (KELVIN): 298.0

REMARK
200
PH: 4.50

REMARK
200
NUMBER OF CRYSTALS USED: 2

REMARK
200

REMARK
200
SYNCHROTRON (Y/N)
: N

REMARK
200
RADIATION SOURCE:
ROTATING ANODE

REMARK
200
BEAMLINE: NULL

REMARK
200
X-RAY GENERATOR MODEL: RIGAKU RU200

REMARK
200
MONOCHROMATIC OR LAUE (M/L): M

REMARK
200
WAVELENGTH OR RANGE (A): 1.5418

REMARK
200
MONOCHROMATOR: GRAPHITE

REMARK
200
OPTICS: GRAPHITE

REMARK
200

REMARK
200
DETECTOR TYPE: IMAGE PLATE

REMARK
200
DETECTOR MANUFACTURER: RIGAKU RAXIS IV

REMARK
200
INTENSITY-INTEGRATION SOFTWARE: CRYSTALCLEAR

(MSC/RIGAKU)

REMARK
200
DATA SCALING SOFTWARE: SCALEPACK

REMARK
200

REMARK
200
NUMBER OF UNIQUE REFLECTIONS: 7013

REMARK
200
RESOLUTION RANGE HIGH (A): 2.750

REMARK
200
RESOLUTION RANGE LOW (A): 100.000

REMARK
200
REJECTION CRITERIA (SIGMA(I)): 0.000

REMARK
200

REMARK
200
OVERALL.

REMARK
200
COMPLETENESS FOR RANGE (%): 93.1

REMARK
200
DATA REDUNDANCY: NULL

REMARK
200
R MERGE (I): 0.07900

REMARK
200
R SYM (I): NULL

REMARK
200
<I/SIGMA(I)> FOR THE DATA SET: 7.3000

REMARK
200

REMARK
200
IN THE HIGHEST RESOLUTION SHELL.

REMARK
200
HIGHEST RESOLUTION SHELL, RANGE HIGH (A): 2.75

REMARK
200
HIGHEST RESOLUTION SHELL, RANGE LOW (A): 2.85

REMARK
200
COMPLETENESS FOR SHELL (%): 96.6

REMARK
200
DATA REDUNDANCY IN SHELL: NULL

REMARK
200
R MERGE FOR SHELL (I): 0.40200

REMARK
200
R SYM FOR SHELL (I): NULL

REMARK
200
<I/SIGMA(I)> FOR SHELL: NULL

REMARK
200

REMARK
200
DIFFRACTION PROTOCOL: SINGLE WAVELENGTH

REMARK
200
METHOD USED TO DETERMINE THE STRUCTURE: MOLECULAR

REPLACEMENT

REMARK
200
SOFTWARE USED: AMORE

REMARK
200
STARTING MODEL: 1N10

REMARK
200

REMARK
200
REMARK: NULL

REMARK
280

REMARK
280
CRYSTAL

REMARK
280
SOLVENT CONTENT, VS (%): 54.03

REMARK
280
MATTHEWS COEFFICIENT, VM (ANGSTROMS**3/DA): 2.68

REMARK
280

REMARK
280
CRYSTALLIZATION CONDITIONS: 15% PEG 4000, 50 MM SODIUM

ACETATE

REMARK
280
AND 0.1M AMMONIUM SULPHATE, PH 4.5, VAPOR DIFFUSION,

HANGING

REMARK
280
DROP, TEMPERATURE 294 K

REMARK
290

REMARK
290
CRYSTALLOGRAPHIC SYMMETRY

REMARK
290
SYMMETRY OPERATORS FOR SPACE GROUP: C 1 2 1

REMARK
290

REMARK
290
SYMOP SYMMETRY

REMARK
290
NNNMMM OPERATOR

REMARK
290
1555
X, Y, Z

REMARK
290
2555
−X, Y, −Z

REMARK
290
3555
½ + X, ½ + Y, Z

REMARK
290
4555
½ − X, ½ + Y, −Z

REMARK
290

REMARK
290
WHERE NNN -> OPERATOR NUMBER

REMARK
290
MMM -> TRANSLATION VECTOR

REMARK
290

REMARK
290
CRYSTALLOGRAPHIC SYMMETRY TRANSFORMATIONS

REMARK
290
THE FOLLOWING TRANSFORMATIONS OPERATE ON THE

ATOM/HETATM

REMARK
290
RECORDS IN THIS ENTRY TO PRODUCE

CRYSTALLOGRAPHICALLY

REMARK
290
RELATED MOLECULES.

REMARK
290
SMTRY1
1
1.000000
0.000000
0.000000
0.00000

REMARK
290
SMTRY2
1
0.000000
1.000000
0.000000
0.00000

REMARK
290
SMTRY3
1
0.000000
0.000000
1.000000
0.00000

REMARK
290
SMTRY1
2
−1.000000
0.000000
0.000000
0.00000

REMARK
290
SMTRY2
2
0.000000
1.000000
0.000000
0.00000

REMARK
290
SMTRY3
2
0.000000
0.000000
−1.000000
0.00000

REMARK
290
SMTRY1
3
1.000000
0.000000
0.000000
56.70400

REMARK
290
SMTRY2
3
0.000000
1.000000
0.000000
22.25600

REMARK
290
SMTRY3
3
0.000000
0.000000
1.000000
0.00000

REMARK
290
SMTRY1
4
−1.000000
0.000000
0.000000
56.70400

REMARK
290
SMTRY2
4
0.000000
1.000000
0.000000
22.25600

REMARK
290
SMTRY3
4
0.000000
0.000000
−1.000000
0.00000

REMARK
290

REMARK
290
REMARK: NULL

REMARK
300

REMARK
300
BIOMOLECULE: 1

REMARK
300
THIS ENTRY CONTAINS THE CRYSTALLOGRAPHIC

ASYMMETRIC UNIT

REMARK
300
WHICH CONSISTS OF 1 CHAIN(S). SEE REMARK 350 FOR

REMARK
300
INFORMATION ON GENERATING THE BIOLOGICAL

MOLECULE(S).

REMARK
350

REMARK
350
GENERATING THE BIOMOLECULE

REMARK
350
COORDINATES FOR A COMPLETE MULTIMER REPRESENTING

THE KNOWN

REMARK
350
BIOLOGICALLY SIGNIFICANT OLIGOMERIZATION STATE OF

THE

REMARK
350
MOLECULE CAN BE GENERATED BY APPLYING BIOMT

TRANSFORMATIONS

REMARK
350
GIVEN BELOW. BOTH NON-CRYSTALLOGRAPHIC AND

REMARK
350
CRYSTALLOGRAPHIC OPERATIONS ARE GIVEN.

REMARK
350

REMARK
350
BIOMOLECULE: 1

REMARK
350
APPLY THE FOLLOWING TO CHAINS: X, A

REMARK
350
BIOMT1
1
1.000000
0.000000
0.000000
0.00000

REMARK
350
BIOMT2
1
0.000000
1.000000
0.000000
0.00000

REMARK
350
BIOMT3
1
0.000000
0.000000
1.000000
0.00000

REMARK
465

REMARK
465
MISSING RESIDUES

REMARK
465
THE FOLLOWING RESIDUES WERE NOT LOCATED IN THE

REMARK
465
EXPERIMENT. (M = MODEL NUMBER; RES = RESIDUE NAME;

C = CHAIN

REMARK
465
IDENTIFIER; SSSEQ = SEQUENCE NUMBER; I = INSERTION CODE.)

REMARK
465

REMARK
465
M RES C SSSEQI

REMARK
465
GLY X 1

REMARK
465
PRO X 2

REMARK
465
PRO X 3

REMARK
500

REMARK
500
GEOMETRY AND STEREOCHEMISTRY

REMARK
500
SUBTOPIC: CLOSE CONTACTS IN SAME ASYMMETRIC UNIT

REMARK
500

REMARK
500
THE FOLLOWING ATOMS ARE IN CLOSE CONTACT.

REMARK
500

REMARK
500
ATM1 RES C SSEQI ATM2 RES C SSEQI

REMARK
500
C3 NAG A 1 C1 FCA A 7 2.10

REMARK
500
O6 MAN A 3 C2 MAN A 5 2.11

REMARK
500
O ASN X 16 N LYS X 18 2.14

REMARK
500

REMARK
500
GEOMETRY AND STEREOCHEMISTRY

REMARK
500
SUBTOPIC: COVALENT BOND LENGTHS

REMARK
500

REMARK
500
THE STEREOCHEMICAL PARAMETERS OF THE FOLLOWING

RESIDUES

REMARK
500
HAVE VALUES WHICH DEVIATE FROM EXPECTED VALUES BY

MORE

REMARK
500
THAN 6*RMSD (M = MODEL NUMBER; RES = RESIDUE NAME;

C = CHAIN

REMARK
500
IDENTIFIER; SSEQ = SEQUENCE NUMBER; I = INSERTION CODE).

REMARK
500

REMARK
500
STANDARD TABLE:

REMARK
500
FORMAT: (10X, I3, 1X, 2(A3, 1X, A1, I4, A1, 1X, A4, 3X), F6.3)

REMARK
500

REMARK
500
EXPECTED VALUES: ENGH AND HUBER, 1991

REMARK
500

REMARK
500
M RES CSSEQI ATM1 RES CSSEQI ATM2 DEVIATION

REMARK
500
ILE X 44 CA ILE X 44 CB 0.052

REMARK
500

REMARK
500
GEOMETRY AND STEREOCHEMISTRY

REMARK
500
SUBTOPIC: COVALENT BOND ANGLES

REMARK
500

REMARK
500
THE STEREOCHEMICAL PARAMETERS OF THE FOLLOWING

RESIDUES

REMARK
500
HAVE VALUES WHICH DEVIATE FROM EXPECTED VALUES BY

MORE

REMARK
500
THAN 6*RMSD (M = MODEL NUMBER; RES = RESIDUE NAME;

C = CHAIN

REMARK
500
IDENTIFIER; SSEQ = SEQUENCE NUMBER; I = INSERTION CODE).

REMARK
500

REMARK
500
STANDARD TABLE:

REMARK
500
FORMAT: (10X, I3, 1X, A3, 1X, A1, I4, A1, 3(1X, A4, 2X), 12X, F5.1)

REMARK
500

REMARK
500
EXPECTED VALUES: ENGH AND HUBER, 1991

REMARK
500

REMARK
500
M RES CSSEQI ATM1 ATM2 ATM3

REMARK
500
ASN X 10 N-CA-C ANGL. DEV. = −11.5 DEGREES

REMARK
500
HYP X 9 CA-C-N ANGL. DEV. = 10.9 DEGREES

REMARK
500
HYP X 9 O-C-N ANGL. DEV. = −12.0 DEGREES

REMARK
500
GLY X 17 N-CA-C ANGL. DEV. = 12.3 DEGREES

REMARK
500
TRP X 19 N-CA-C ANGL. DEV. = 10.7 DEGREES

REMARK
500
ILE X 44 N-CA-C ANGL. DEV. = −10.3 DEGREES

REMARK
500
CYS X 73 CA-CB-SG ANGL. DEV. = −9.8 DEGREES

REMARK
500
CYS X 128 N-CA-C ANGL. DEV. = −11.2 DEGREES

REMARK
500

REMARK
500
GEOMETRY AND STEREOCHEMISTRY

REMARK
500
SUBTOPIC: TORSION ANGLES

REMARK
500

REMARK
500
TORSION ANGLES OUTSIDE THE EXPECTED RAMACHANDRAN

REGIONS:

REMARK
500
(M = MODEL NUMBER; RES = RESIDUE NAME; C = CHAIN

IDENTIFIER;

REMARK
500
SSEQ = SEQUENCE NUMBER; I = INSERTION CODE).

REMARK
500

REMARK
500
STANDARD TABLE:

REMARK
500
FORMAT: (10X, I3, 1X, A3, 1X, A1, I4, A1, 4X, F7.2, 3X, F7.2)

REMARK
500

REMARK
500
M RES CSSEQI PSI PHI

REMARK
500
ASN X 10 113.05 138.93

REMARK
500
CYS X 156 −117.96 −158.76

REMARK
500
ALA X 162 111.35 121.14

REMARK
500
ASP X 170 −97.85 81.46

REMARK
500
LEU X 183 127.07 92.22

REMARK
500

REMARK
500
GEOMETRY AND STEREOCHEMISTRY

REMARK
500
SUBTOPIC: NON-CIS, NON-TRANS

REMARK
500

REMARK
500
THE FOLLOWING PEPTIDE BONDS DEVIATE SIGNIFICANTLY

FROM BOTH

REMARK
500
CIS AND TRANS CONFORMATION. CIS BONDS, IF ANY, ARE

LISTED

REMARK
500
ON CISPEP RECORDS. TRANS IS DEFINED AS 180 +/− 30 AND

REMARK
500
CIS IS DEFINED AS 0 +/− 30 DEGREES.

REMARK
500
MODEL OMEGA

REMARK
500
GLY X 8 HYP X 9 60.63

REMARK
500
HYP X 9 ASN X 10 −64.54

REMARK
525

REMARK
525
SOLVENT

REMARK
525
THE FOLLOWING SOLVENT MOLECULES LIE FARTHER THAN

EXPECTED

REMARK
525
FROM THE PROTEIN OR NUCLEIC ACID MOLECULE AND MAY

BE

REMARK
525
ASSOCIATED WITH A SYMMETRY RELATED MOLECULE

(M = MODEL

REMARK
525
NUMBER; RES = RESIDUE NAME; C = CHAIN IDENTIFIER;

SSEQ = SEQUENCE

REMARK
525
NUMBER; I = INSERTION CODE):

REMARK
525

REMARK
525
M RES CSSEQI

REMARK
525
HOH 7 DISTANCE = 5.93 ANGSTROMS

REMARK
525
HOH 17 DISTANCE = 5.21 ANGSTROMS

REMARK
900

REMARK
900
RELATED ENTRIES

REMARK
900
RELATED ID: 1N10 RELATED DB: PDB

REMARK
900
CRYSTAL STRUCTURE OF PHL P 1, A MAJOR TIMOTHY GRASS

POLLEN

REMARK
900
ALLERGEN

DBREF 2HCZ X 1 245 UNP P58738 EXB1A_MAIZE 25 269

SEQADV 2HCZ HYP X 9 UNP P58738 PRO 33 MODIFIED RESIDUE

SEQRES
1 X
245
GLY PRO PRO LYS VAL PRO PRO GLY HYP ASN ILE THR

THR

SEQRES
2 X
245
ASN TYR ASN GLY LYS TRP LEU THR ALA ARG ALA THR

TRP

SEQRES
3 X
245
TYR GLY GLN PRO ASN GLY ALA GLY ALA PRO ASP ASN

GLY

SEQRES
4 X
245
GLY ALA CYS GLY ILE LYS ASN VAL ASN LEU PRO PRO

TYR

SEQRES
5 X
245
SER GLY MET THR ALA CYS GLY ASN VAL PRO ILE PHE

LYS

SEQRES
6 X
245
ASP GLY LYS GLY CYS GLY SER CYS TYR GLU VAL ARG

CYS

SEQRES
7 X
245
LYS GLU LYS PRO GLU CYS SER GLY ASN PRO VAL THR

VAL

SEQRES
8 X
245
TYR ILE THR ASP MET ASN TYR GLU PRO ILE ALA PRO TYR

SEQRES
9 X
245
HIS PHE ASP LEU SER GLY LYS ALA PHE GLY SER LEU ALA

SEQRES
10 X
245
LYS PRO GLY LEU ASN ASP LYS ILE ARG HIS CYS GLY ILE

SEQRES
11 X
245
MET ASP VAL GLU PHE ARG ARG VAL ARG CYS LYS TYR

PRO

SEQRES
12 X
245
ALA GLY GLN LYS ILE VAL PHE HIS ILE GLU LYS GLY CYS

SEQRES
13 X
245
ASN PRO ASN TYR LEU ALA VAL LEU VAL LYS TYR VAL

ALA

SEQRES
14 X
245
ASP ASP GLY ASP ILE VAL LEU MET GLU ILE GLN ASP LYS

SEQRES
15 X
245
LEU SER ALA GLU TRP LYS PRO MET LYS LEU SER TRP

GLY

SEQRES
16 X
245
ALA ILE TRP ARG MET ASP THR ALA LYS ALA LEU LYS

GLY

SEQRES
17 X
245
PRO PHE SER ILE ARG LEU THR SER GLU SER GLY LYS LYS

SEQRES
18 X
245
VAL ILE ALA LYS ASP VAL ILE PRO ALA ASN TRP ARG PRO

SEQRES
19 X
245
ASP ALA VAL TYR THR SER ASN VAL GLN PHE TYR

MODRES 2HCZ HYP X 9 PRO 4-HYDROXYPROLINE

MODRES 2HCZ ASN X 10 ASN GLYCOSYLATION SITE

HET
HYP
X
9
8

HET
NAG
A
1
14

HET
NAG
A
2
14

HET
MAN
A
3
11

HET
MAN
A
4
11

HET
XYS
A
6
9

HET
MAN
A
5
11

HET
FCA
A
7
10

HETNAM
HYP 4-HYDROXYPROLINE

HETNAM
NAG N-ACETYL-D-GLUCOSAMINE

HETNAM
MAN ALPHA-D-MANNOSE

HETNAM
XYS XYLOPYRANOSE

HETNAM
FCA ALPHA-D-FUCOSE

HETSYN
HYP HYDROXYPROLTNE

HETSYN
NAG NAG

FORMUL
1 HYP C5 H9 N O3

FORMUL
2 NAG 2(C8 H15 N O6)

FORMUL
2 MAN 3(C6 H12 O6)

FORMUL
2 XYS C5 H10 O5

FORMUL
4 FCA C6 H12 O5

FORMUL
5 HOH *17(H2 O)

HELIX
1
1 ASN X 60 LYS X 65 1
6

HELIX
2
2 ASP X 66 LYS X 68 5
3

HELIX
3
3 SER X 109 LEU X 116 1
8

HELIX
4
4 LEU X 121 ARG X 126 1
6

SHEET
1
A 7 LEU X 20 TRP X 26 0

SHEET
2
A 7 HIS X 105 LEU X 108 1 O LEU X 108 N THR X 25

SHEET
3
A 7 THR X 56 GLY X 59 −1 N CYS X 58 O ASP X 107

SHEET
4
A 7 VAL X 89 MET X 96 1 O ASP X 95 N ALA X 57

SHEET
5
A 7 CYS X 73 ARG X 77 −1 N VAL X 76 O VAL X 89

SHEET
6
A 7 MET X 131 VAL X 138 −1 O GLU X 134 N ARG X 77

SHEET
7
A 7 LEU X 20 TRP X 26 −1 N ALA X 24 O MET X 131

SHEET
1
B 5 LYS X 191 TRP X 194 0

SHEET
2
B 5 ILE X 197 ARG X 199 −1 O ARG X 199 N LYS X 191

SHEET
3
B 5 VAL X 163 LYS X 166 −1 N VAL X 163 O TRP X 198

SHEET
4
B 5 VAL X 149 ILE X 152 −1 N HIS X 151 O LEU X 164

SHEET
5
B 5 VAL X 237 THR X 239 −1 O TYR X 238 N PHE X 150

SHEET
1
C 3 ILE X 174 GLU X 178 0

SHEET
2
C 3 ILE X 212 SER X 216 −1 O ARG X 213 N GLU X 178

SHEET
3
C 3 LYS X 221 ALA X 224 −1 O ALA X 224 N ILE X 212

SSBOND
1
CYS X 42 CYS X 70

SSBOND
2
CYS X 73 CYS X 140

SSB0ND
3
CYS X 78 CYS X 84

LINK
ND2 ASN X 10
C1 NAG A 1

LINK
O4 NAG A 1
C1 NAG A 2

LINK
O4 NAG A 2
C1 MAN A 3

LINK
O2 MAN A 3
C1 XYS A 6

LINK
O6 MAN A 3
C1 MAN A 5

LINK
O3 NAG A 1
C1 FCA A 7

LINK
O3 MAN A 3
C1 MAN A 4

CISPEP 1 PRO X 50 PRO X 51 0 −0.15

CRYST1 113.408 44.512 69.467 90.00 124.64 90.00 C 1 2 1 4

ORIGX1
1.000000
0.000000
0.000000
0.00000

ORIGX2
0.000000
1.000000
0.000000
0.00000

ORIGX3
0.000000
0.000000
1.000000
0.00000

SCALE1
0.008818
0.000000
0.006092
0.00000

SCALE2
0.000000
0.022466
0.000000
0.00000

SCALE3
0.000000
0.000000
0.017497
0.00000

ATOM
1
N
LYS
X
4
−11.044
−7.638
34.841
1.00
92.39
N

ATOM
2
CA
LYS
X
4
−9.973
−7.749
33.810
1.00
92.39
C

ATOM
3
C
LYS
X
4
−10.533
−7.543
32.405
1.00
92.39
C

ATOM
4
O
LYS
X
4
−11.535
−8.149
32.032
1.00
92.39
O

ATOM
5
CB
LYS
X
4
−9.313
−9.126
33.897
1.00
90.09
C

ATOM
6
CG
LYS
X
4
−8.209
−9.354
32.881
1.00
90.09
C

ATOM
7
CD
LYS
X
4
−7.712
−10.784
32.940
1.00
90.09
C

ATOM
8
CE
LYS
X
4
−6.665
−11.045
31.875
1.00
90.09
C

ATOM
9
NZ
LYS
X
4
−6.247
−12.474
31.854
1.00
90.09
N

ATOM
10
N
VAL
X
5
−9.882
−6.684
31.629
1.00
61.66
N

ATOM
11
CA
VAL
X
5
−10.323
−6.418
30.267
1.00
61.66
C

ATOM
12
C
VAL
X
5
−9.453
−7.136
29.242
1.00
61.66
C

ATOM
13
O
VAL
X
5
−8.236
−6.963
29.217
1.00
61.66
O

ATOM
14
CB
VAL
X
5
−10.316
−4.906
29.959
1.00
43.90
C

ATOM
15
CG1
VAL
X
5
−10.042
−4.672
28.489
1.00
43.90
C

ATOM
16
CG2
VAL
X
5
−11.659
−4.301
30.332
1.00
43.90
C

ATOM
17
N
PRO
X
6
−10.079
−7.962
28.388
1.00
62.91
N

ATOM
18
CA
PRO
X
6
−9.422
−8.739
27.329
1.00
62.91
C

ATOM
19
C
PRO
X
6
−9.096
−7.921
26.067
1.00
62.91
C

ATOM
20
O
PRO
X
6
−9.963
−7.256
25.501
1.00
62.91
O

ATOM
21
CB
PRO
X
6
−10.430
−9.854
27.056
1.00
60.98
C

ATOM
22
CG
PRO
X
6
−11.740
−9.160
27.261
1.00
60.98
C

ATOM
23
CD
PRO
X
6
−11.493
−8.361
28.528
1.00
60.98
C

ATOM
24
N
PRO
X
7
−7.837
−7.987
25.605
1.00
44.83
N

ATOM
25
CA
PRO
X
7
−7.310
−7.287
24.426
1.00
44.83
C

ATOM
26
C
PRO
X
7
−8.084
−7.529
23.140
1.00
44.83
C

ATOM
27
O
PRO
X
7
−8.671
−8.583
22.968
1.00
44.83
O

ATOM
28
CB
PRO
X
7
−5.872
−7.794
24.336
1.00
45.78
C

ATOM
29
CG
PRO
X
7
−5.955
−9.164
24.956
1.00
45.78
C

ATOM
30
CD
PRO
X
7
−6.831
−8.916
26.148
1.00
45.78
C

ATOM
31
N
GLY
X
8
−8.054
−6.535
22.254
1.00
94.04
N

ATOM
32
CA
GLY
X
8
−8.745
−6.563
20.971
1.00
94.04
C

ATOM
33
C
GLY
X
8
−9.377
−7.843
20.445
1.00
94.04
C

ATOM
34
O
GLY
X
8
−10.249
−8.411
21.101
1.00
94.04
O

HETATM
35
N
HYP
X
9
−9.033
−8.354
19.261
1.00
57.41
N

HETATM
36
CA
HYP
X
9
−7.613
−8.797
18.932
1.00
57.41
C

HETATM
37
C
HYP
X
9
−6.643
−7.615
19.045
1.00
57.41
C

HETATM
38
O
HYP
X
9
−6.058
−7.717
20.111
1.00
57.41
O

HETATM
39
CB
HYP
X
9
−7.650
−9.469
17.547
1.00
57.41
C

HETATM
40
CG
HYP
X
9
−8.931
−9.087
16.923
1.00
57.41
C

HETATM
41
CD
HYP
X
9
−9.620
−8.151
17.908
1.00
57.41
C

HETATM
42
OD1
HYP
X
9
−9.742
−10.250
16.748
1.00
57.41
O

ATOM
43
N
ASN
X
10
−5.825
−7.147
18.092
1.00
65.18
N

ATOM
44
CA
ASN
X
10
−4.661
−7.776
17.413
1.00
65.18
C

ATOM
45
C
ASN
X
10
−4.911
−7.316
16.006
1.00
65.18
C

ATOM
46
O
ASN
X
10
−5.894
−7.692
15.386
1.00
65.18
O

ATOM
47
CB
ASN
X
10
−4.637
−9.290
17.513
1.00
42.31
C

ATOM
48
CG
ASN
X
10
−3.220
−9.895
17.664
1.00
42.31
C

ATOM
49
OD1
ASN
X
10
−2.478
−10.011
16.693
1.00
42.31
O

ATOM
50
ND2
ASN
X
10
−2.872
−10.219
18.917
1.00
42.31
N

ATOM
51
N
ILE
X
11
−4.019
−6.463
15.524
1.00
32.17
N

ATOM
52
CA
ILE
X
11
−4.162
−5.836
14.228
1.00
32.17
C

ATOM
53
C
ILE
X
11
−3.131
−6.269
13.218
1.00
32.17
C

ATOM
54
O
ILE
X
11
−1.929
−6.249
13.471
1.00
32.17
O

ATOM
55
CB
ILE
X
11
−4.196
−4.291
14.444
1.00
23.05
C

ATOM
56
CG1
ILE
X
11
−5.539
−3.944
15.077
1.00
23.05
C

ATOM
57
CG2
ILE
X
11
−4.015
−3.525
13.158
1.00
23.05
C

ATOM
58
CD1
ILE
X
11
−5.648
−2.564
15.569
1.00
23.05
C

ATOM
59
N
THR
X
12
−3.636
−6.670
12.059
1.00
42.32
N

ATOM
60
CA
THR
X
12
−2.805
−7.160
10.974
1.00
42.32
C

ATOM
61
C
THR
X
12
−2.568
−6.124
9.907
1.00
42.32
C

ATOM
62
O
THR
X
12
−3.041
−4.994
10.004
1.00
42.32
O

ATOM
63
CB
THR
X
12
−3.449
−8.365
10.314
1.00
51.84
C

ATOM
64
OG1
THR
X
12
−4.786
−8.024
9.932
1.00
51.84
O

ATOM
65
CG2
THR
X
12
−3.484
−9.540
11.277
1.00
51.84
C

ATOM
66
N
THR
X
13
−1.826
−6.521
8.882
1.00
31.52
N

ATOM
67
CA
THR
X
13
−1.532
−5.620
7.793
1.00
31.52
C

ATOM
68
C
THR
X
13
−2.739
−5.437
6.868
1.00
31.52
C

ATOM
69
O
THR
X
13
−2.620
−4.935
5.754
1.00
31.52
O

ATOM
70
CB
THR
X
13
−0.353
−6.120
7.003
1.00
31.48
C

ATOM
71
OG1
THR
X
13
−0.615
−7.453
6.563
1.00
31.48
O

ATOM
72
CG2
THR
X
13
0.889
−6.096
7.862
1.00
31.48
C

ATOM
73
N
ASN
X
14
−3.909
−5.827
7.346
1.00
39.41
N

ATOM
74
CA
ASN
X
14
−5.118
−5.663
6.567
1.00
39.41
C

ATOM
75
C
ASN
X
14
−5.551
−4.194
6.652
1.00
39.41
C

ATOM
76
O
ASN
X
14
−6.186
−3.785
7.621
1.00
39.41
O

ATOM
77
CB
ASN
X
14
−6.210
−6.583
7.122
1.00
75.84
C

ATOM
78
CG
ASN
X
14
−7.544
−6.392
6.433
1.00
75.84
C

ATOM
79
OD1
ASN
X
14
−7.617
−6.311
5.205
1.00
75.84
O

ATOM
80
ND2
ASN
X
14
−8.613
−6.329
7.222
1.00
75.84
N

ATOM
81
N
TYR
X
15
−5.205
−3.398
5.645
1.00
34.29
N

ATOM
82
CA
TYR
X
15
−5.574
−1.986
5.648
1.00
34.29
C

ATOM
83
C
TYR
X
15
−7.024
−1.708
5.222
1.00
34.29
C

ATOM
84
O
TYR
X
15
−7.256
−1.197
4.135
1.00
34.29
O

ATOM
85
CB
TYR
X
15
−4.605
−1.202
4.750
1.00
28.79
C

ATOM
86
CG
TYR
X
15
−3.152
−1.309
5.199
1.00
28.79
C

ATOM
87
CD1
TYR
X
15
−2.227
−2.065
4.487
1.00
28.79
C

ATOM
88
CD2
TYR
X
15
−2.724
−0.689
6.381
1.00
28.79
C

ATOM
89
CE1
TYR
X
15
−0.927
−2.206
4.941
1.00
28.79
C

ATOM
90
CE2
TYR
X
15
−1.426
−0.822
6.842
1.00
28.79
C

ATOM
91
CZ
TYR
X
15
−0.533
−1.578
6.127
1.00
28.79
C

ATOM
92
OH
TYR
X
15
0.752
−1.708
6.609
1.00
28.79
O

ATOM
93
N
ASN
X
16
−7.998
−2.021
6.081
1.00
45.44
N

ATOM
94
CA
ASN
X
16
−9.414
−1.793
5.758
1.00
45.44
C

ATOM
95
C
ASN
X
16
−9.935
−0.433
6.215
1.00
45.44
C

ATOM
96
O
ASN
X
16
−10.921
−0.343
6.942
1.00
45.44
O

ATOM
97
CB
ASN
X
16
−10.301
−2.887
6.363
1.00
49.59
C

ATOM
98
CG
ASN
X
16
−10.458
−2.758
7.868
1.00
49.59
C

ATOM
99
OD1
ASN
X
16
−9.623
−3.225
8.626
1.00
49.59
O

ATOM
100
ND2
ASN
X
16
−11.534
−2.116
8.301
1.00
49.59
N

ATOM
101
N
GLY
X
17
−9.257
0.614
5.763
1.00
44.10
N

ATOM
102
CA
GLY
X
17
−9.595
1.992
6.084
1.00
44.10
C

ATOM
103
C
GLY
X
17
−10.736
2.415
6.993
1.00
44.10
C

ATOM
104
O
GLY
X
17
−11.173
3.562
6.900
1.00
44.10
O

ATOM
105
N
LYS
X
18
−11.240
1.558
7.871
1.00
46.12
N

ATOM
106
CA
LYS
X
18
−12.306
2.036
8.730
1.00
46.12
C

ATOM
107
C
LYS
X
18
−12.032
1.996
10.222
1.00
46.12
C

ATOM
108
O
LYS
X
18
−11.373
1.103
10.737
1.00
46.12
O

ATOM
109
CB
LYS
X
18
−13.639
1.380
8.374
1.00
99.00
C

ATOM
110
CG
LYS
X
18
−14.290
2.045
7.150
1.00
99.00
C

ATOM
111
CD
LYS
X
18
−14.283
3.571
7.291
1.00
99.00
C

ATOM
112
CE
LYS
X
18
−14.408
4.267
5.946
1.00
99.00
C

ATOM
113
NZ
LYS
X
18
−14.104
5.729
6.041
1.00
99.00
N

ATOM
114
N
TRP
X
19
−12.568
3.005
10.894
1.00
43.83
N

ATOM
115
CA
TRP
X
19
−12.366
3.248
12.308
1.00
43.83
C

ATOM
116
C
TRP
X
19
−12.793
2.257
13.354
1.00
43.83
C

ATOM
117
O
TRP
X
19
−13.855
1.659
13.283
1.00
43.83
O

ATOM
118
CB
TRP
X
19
−12.938
4.620
12.638
1.00
49.74
C

ATOM
119
CG
TRP
X
19
−12.492
5.613
11.625
1.00
49.74
C

ATOM
120
CD1
TRP
X
19
−13.056
5.850
10.398
1.00
49.74
C

ATOM
121
CD2
TRP
X
19
−11.315
6.422
11.688
1.00
49.74
C

ATOM
122
NE1
TRP
X
19
−12.295
6.754
9.694
1.00
49.74
N

ATOM
123
CE2
TRP
X
19
−11.222
7.122
10.462
1.00
49.74
C

ATOM
124
CE3
TRP
X
19
−10.328
6.624
12.660
1.00
49.74
C

ATOM
125
CZ2
TRP
X
19
−10.179
8.009
10.186
1.00
49.74
C

ATOM
126
CZ3
TRP
X
19
−9.291
7.505
12.386
1.00
49.74
C

ATOM
127
CH2
TRP
X
19
−9.225
8.188
11.158
1.00
49.74
C

ATOM
128
N
LEU
X
20
−11.924
2.115
14.345
1.00
31.46
N

ATOM
129
CA
LEU
X
20
−12.138
1.231
15.470
1.00
31.46
C

ATOM
130
C
LEU
X
20
−12.099
2.090
16.730
1.00
31.46
C

ATOM
131
O
LEU
X
20
−11.491
3.161
16.740
1.00
31.46
O

ATOM
132
CB
LEU
X
20
−11.020
0.202
15.530
1.00
26.96
C

ATOM
133
CG
LEU
X
20
−10.943
−0.813
14.404
1.00
26.96
C

ATOM
134
CD1
LEU
X
20
−9.602
−1.516
14.413
1.00
26.96
C

ATOM
135
CD2
LEU
X
20
−12.078
−1.792
14.584
1.00
26.96
C

ATOM
136
N
THR
X
21
−12.742
1.617
17.790
1.00
52.90
N

ATOM
137
CA
THR
X
21
−12.760
2.347
19.046
1.00
52.90
C

ATOM
138
C
THR
X
21
−11.534
1.975
19.873
1.00
52.90
C

ATOM
139
O
THR
X
21
−10.907
0.939
19.650
1.00
52.90
O

ATOM
140
CB
THR
X
21
−14.023
2.032
19.853
1.00
51.86
C

ATOM
141
OG1
THR
X
21
−15.171
2.252
19.033
1.00
51.86
O

ATOM
142
CG2
THR
X
21
−14.128
2.940
21.062
1.00
51.86
C

ATOM
143
N
ALA
X
22
−11.194
2.834
20.824
1.00
55.16
N

ATOM
144
CA
ALA
X
22
−10.050
2.612
21.686
1.00
55.16
C

ATOM
145
C
ALA
X
22
−9.981
3.743
22.696
1.00
55.16
C

ATOM
146
O
ALA
X
22
−10.505
4.825
22.450
1.00
55.16
O

ATOM
147
CB
ALA
X
22
−8.781
2.579
20.858
1.00
9.51
C

ATOM
148
N
ARG
X
23
−9.350
3.488
23.837
1.00
59.92
N

ATOM
149
CA
ARG
X
23
−9.205
4.517
24.853
1.00
59.92
C

ATOM
150
C
ARG
X
23
−7.787
5.051
24.811
1.00
59.92
C

ATOM
151
O
ARG
X
23
−6.820
4.289
24.771
1.00
59.92
O

ATOM
152
CB
ARG
X
23
−9.533
3.973
26.246
1.00
77.51
C

ATOM
153
CG
ARG
X
23
−11.019
4.054
26.577
1.00
77.51
C

ATOM
154
CD
ARG
X
23
−11.285
4.021
28.083
1.00
77.51
C

ATOM
155
NE
ARG
X
23
−12.625
4.511
28.420
1.00
77.51
N

ATOM
156
CZ
ARG
X
23
−13.048
5.757
28.215
1.00
77.51
C

ATOM
157
NH1
ARG
X
23
−12.243
6.661
27.672
1.00
77.51
N

ATOM
158
NH2
ARG
X
23
−14.283
6.102
28.552
1.00
77.51
N

ATOM
159
N
ALA
X
24
−7.659
6.369
24.805
1.00
44.47
N

ATOM
160
CA
ALA
X
24
−6.341
6.966
24.748
1.00
44.47
C

ATOM
161
C
ALA
X
24
−6.021
7.717
26.014
1.00
44.47
C

ATOM
162
O
ALA
X
24
−6.913
8.109
26.760
1.00
44.47
O

ATOM
163
CB
ALA
X
24
−6.253
7.906
23.564
1.00
47.78
C

ATOM
164
N
THR
X
25
−4.732
7.894
26.255
1.00
50.03
N

ATOM
165
CA
THR
X
25
−4.239
8.644
27.402
1.00
50.03
C

ATOM
166
C
THR
X
25
−2.843
9.018
26.977
1.00
50.03
C

ATOM
167
O
THR
X
25
−2.365
8.568
25.931
1.00
50.03
O

ATOM
168
CB
THR
X
25
−4.123
7.808
28.701
1.00
22.06
C

ATOM
169
OG1
THR
X
25
−3.065
6.849
28.560
1.00
22.06
O

ATOM
170
CG2
THR
X
25
−5.435
7.106
29.015
1.00
22.06
C

ATOM
171
N
TRP
X
26
−2.181
9.833
27.778
1.00
39.67
N

ATOM
172
CA
TRP
X
26
−0.832
10.237
27.435
1.00
39.67
C

ATOM
173
C
TRP
X
26
0.069
10.120
28.645
1.00
39.67
C

ATOM
174
O
TRP
X
26
−0.404
10.061
29.777
1.00
39.67
O

ATOM
175
CB
TRP
X
26
−0.825
11.673
26.900
1.00
46.69
C

ATOM
176
CG
TRP
X
26
−1.363
12.699
27.859
1.00
46.69
C

ATOM
177
CD1
TRP
X
26
−2.668
12.895
28.210
1.00
46.69
C

ATOM
178
CD2
TRP
X
26
−0.605
13.684
28.562
1.00
46.69
C

ATOM
179
NE1
TRP
X
26
−2.767
13.948
29.086
1.00
46.69
N

ATOM
180
CE2
TRP
X
26
−1.515
14.451
29.318
1.00
46.69
C

ATOM
181
CE3
TRP
X
26
0.760
13.995
28.624
1.00
46.69
C

ATOM
182
CZ2
TRP
X
26
−1.111
15.511
30.122
1.00
46.69
C

ATOM
183
CZ3
TRP
X
26
1.165
15.045
29.421
1.00
46.69
C

ATOM
184
CH2
TRP
X
26
0.230
15.795
30.162
1.00
46.69
C

ATOM
185
N
TYR
X
27
1.368
10.083
28.402
1.00
45.95
N

ATOM
186
CA
TYR
X
27
2.314
9.957
29.486
1.00
45.95
C

ATOM
187
C
TYR
X
27
3.535
10.801
29.163
1.00
45.95
C

ATOM
188
O
TYR
X
27
3.703
11.238
28.018
1.00
45.95
O

ATOM
189
CB
TYR
X
27
2.719
8.497
29.644
1.00
30.14
C

ATOM
190
CG
TYR
X
27
3.577
7.999
28.511
1.00
30.14
C

ATOM
191
CD1
TYR
X
27
3.085
7.956
27.206
1.00
30.14
C

ATOM
192
CD2
TYR
X
27
4.895
7.583
28.737
1.00
30.14
C

ATOM
193
CE1
TYR
X
27
3.891
7.509
26.138
1.00
30.14
C

ATOM
194
CE2
TYR
X
27
5.710
7.129
27.677
1.00
30.14
C

ATOM
195
CZ
TYR
X
27
5.197
7.098
26.378
1.00
30.14
C

ATOM
196
OH
TYR
X
27
5.981
6.670
25.328
1.00
30.14
O

ATOM
197
N
GLY
X
28
4.383
11.020
30.170
1.00
44.81
N

ATOM
198
CA
GLY
X
28
5.584
11.820
29.980
1.00
44.81
C

ATOM
199
C
GLY
X
28
5.305
13.312
30.079
1.00
44.81
C

ATOM
200
O
GLY
X
28
4.337
13.730
30.722
1.00
44.81
O

ATOM
201
N
GLN
X
29
6.148
14.122
29.443
1.00
65.60
N

ATOM
202
CA
GLN
X
29
5.943
15.565
29.470
1.00
65.60
C

ATOM
203
C
GLN
X
29
4.793
15.924
28.532
1.00
65.60
C

ATOM
204
O
GLN
X
29
4.520
15.209
27.570
1.00
65.60
O

ATOM
205
CB
GLN
X
29
7.221
16.299
29.066
1.00
87.69
C

ATOM
206
CG
GLN
X
29
7.660
17.361
30.078
1.00
87.69
C

ATOM
207
CD
GLN
X
29
7.957
16.790
31.467
1.00
87.69
C

ATOM
208
OE1
GLN
X
29
7.073
16.255
32.141
1.00
87.69
O

ATOM
209
NE2
GLN
X
29
9.209
16.905
31.897
1.00
87.69
N

ATOM
210
N
PRO
X
30
4.108
17.045
28.797
1.00
50.97
N

ATOM
211
CA
PRO
X
30
2.973
17.495
27.984
1.00
50.97
C

ATOM
212
C
PRO
X
30
3.313
17.904
26.555
1.00
50.97
C

ATOM
213
O
PRO
X
30
2.447
17.913
25.690
1.00
50.97
O

ATOM
214
CB
PRO
X
30
2.403
18.661
28.797
1.00
69.17
C

ATOM
215
CG
PRO
X
30
2.977
18.465
30.189
1.00
69.17
C

ATOM
216
CD
PRO
X
30
4.360
17.994
29.890
1.00
69.17
C

ATOM
217
N
ASN
X
31
4.568
18.260
26.317
1.00
33.36
N

ATOM
218
CA
ASN
X
31
5.017
18.658
24.991
1.00
33.36
C

ATOM
219
C
ASN
X
31
6.219
17.812
24.633
1.00
33.36
C

ATOM
220
O
ASN
X
31
7.083
18.221
23.854
1.00
33.36
O

ATOM
221
CB
ASN
X
31
5.394
20.142
24.963
1.00
78.89
C

ATOM
222
CG
ASN
X
31
4.264
21.022
24.463
1.00
78.89
C

ATOM
223
OD1
ASN
X
31
3.174
21.040
25.036
1.00
78.89
O

ATOM
224
ND2
ASN
X
31
4.519
21.756
23.386
1.00
78.89
N

ATOM
225
N
GLY
X
32
6.268
16.623
25.221
1.00
72.27
N

ATOM
226
CA
GLY
X
32
7.360
15.711
24.954
1.00
72.27
C

ATOM
227
C
GLY
X
32
6.895
14.567
24.077
1.00
72.27
C

ATOM
228
O
GLY
X
32
5.734
14.515
23.665
1.00
72.27
O

ATOM
229
N
ALA
X
33
7.808
13.647
23.795
1.00
50.75
N

ATOM
230
CA
ALA
X
33
7.501
12.495
22.966
1.00
50.75
C

ATOM
231
C
ALA
X
33
7.306
11.232
23.814
1.00
50.75
C

ATOM
232
O
ALA
X
33
7.839
10.174
23.493
1.00
50.75
O

ATOM
233
CB
ALA
X
33
8.622
12.287
21.950
1.00
41.40
C

ATOM
234
N
GLY
X
34
6.550
11.348
24.901
1.00
51.63
N

ATOM
235
CA
GLY
X
34
6.312
10.198
25.758
1.00
51.63
C

ATOM
236
C
GLY
X
34
7.402
9.870
26.774
1.00
51.63
C

ATOM
237
O
GLY
X
34
7.419
10.402
27.894
1.00
51.63
O

ATOM
238
N
ALA
X
35
8.313
8.980
26.388
1.00
81.75
N

ATOM
239
CA
ALA
X
35
9.407
8.570
27.263
1.00
81.75
C

ATOM
240
C
ALA
X
35
10.295
9.745
27.653
1.00
81.75
C

ATOM
241
O
ALA
X
35
10.804
10.460
26.792
1.00
81.75
O

ATOM
242
CB
ALA
X
35
10.241
7.502
26.581
1.00
76.43
C

ATOM
243
N
PRO
X
36
10.487
9.963
28.964
1.00
39.52
N

ATOM
244
CA
PRO
X
36
11.322
11.059
29.458
1.00
39.52
C

ATOM
245
C
PRO
X
36
12.663
11.134
28.730
1.00
39.52
C

ATOM
246
O
PRO
X
36
13.208
12.218
28.512
1.00
39.52
O

ATOM
247
CB
PRO
X
36
11.467
10.720
30.930
1.00
53.57
C

ATOM
248
CG
PRO
X
36
10.105
10.173
31.256
1.00
53.57
C

ATOM
249
CD
PRO
X
36
9.841
9.250
30.085
1.00
53.57
C

ATOM
250
N
ASP
X
37
13.172
9.970
28.338
1.00
47.65
N

ATOM
251
CA
ASP
X
37
14.447
9.865
27.624
1.00
47.65
C

ATOM
252
C
ASP
X
37
14.282
9.995
26.111
1.00
47.65
C

ATOM
253
O
ASP
X
37
15.265
10.016
25.369
1.00
47.65
O

ATOM
254
CB
ASP
X
37
15.106
8.519
27.930
1.00
65.15
C

ATOM
255
CG
ASP
X
37
14.152
7.350
27.754
1.00
65.15
C

ATOM
256
OD1
ASP
X
37
14.633
6.206
27.628
1.00
65.15
O

ATOM
257
OD2
ASP
X
37
12.919
7.572
27.752
1.00
65.15
O

ATOM
258
N
ASN
X
38
13.033
10.078
25.664
1.00
73.67
N

ATOM
259
CA
ASN
X
38
12.723
10.181
24.246
1.00
73.67
C

ATOM
260
C
ASN
X
38
13.202
8.932
23.514
1.00
73.67
C

ATOM
261
O
ASN
X
38
13.939
8.999
22.526
1.00
73.67
O

ATOM
262
CB
ASN
X
38
13.361
11.428
23.642
1.00
36.05
C

ATOM
263
CG
ASN
X
38
12.766
12.705
24.190
1.00
36.05
C

ATOM
264
OD1
ASN
X
38
11.569
12.785
24.481
1.00
36.05
O

ATOM
265
ND2
ASN
X
38
13.599
13.722
24.318
1.00
36.05
N

ATOM
266
N
GLY
X
39
12.768
7.790
24.031
1.00
36.67
N

ATOM
267
CA
GLY
X
39
13.106
6.500
23.465
1.00
36.67
C

ATOM
268
C
GLY
X
39
11.937
5.627
23.860
1.00
36.67
C

ATOM
269
O
GLY
X
39
10.855
6.154
24.139
1.00
36.67
O

ATOM
270
N
GLY
X
40
12.129
4.313
23.915
1.00
17.90
N

ATOM
271
CA
GLY
X
40
11.015
3.462
24.293
1.00
17.90
C

ATOM
272
C
GLY
X
40
11.279
1.982
24.121
1.00
17.90
C

ATOM
273
O
GLY
X
40
12.359
1.547
23.681
1.00
17.90
O

ATOM
274
N
ALA
X
41
10.260
1.205
24.465
1.00
32.57
N

ATOM
275
CA
ALA
X
41
10.332
−0.233
24.390
1.00
32.57
C

ATOM
276
C
ALA
X
41
10.851
−0.749
23.067
1.00
32.57
C

ATOM
277
O
ALA
X
41
11.387
−1.848
23.023
1.00
32.57
O

ATOM
278
CB
ALA
X
41
8.985
−0.817
24.675
1.00
38.27
C

ATOM
279
N
CYS
X
42
10.706
0.025
21.991
1.00
38.80
N

ATOM
280
CA
CYS
X
42
11.182
−0.422
20.678
1.00
38.80
C

ATOM
281
C
CYS
X
42
12.672
−0.248
20.500
1.00
38.80
C

ATOM
282
O
CYS
X
42
13.289
−0.915
19.673
1.00
38.80
O

ATOM
283
CB
CYS
X
42
10.464
0.313
19.539
1.00
32.69
C

ATOM
284
SG
CYS
X
42
8.700
−0.098
19.380
1.00
32.69
S

ATOM
285
N
GLY
X
43
13.257
0.665
21.259
1.00
43.16
N

ATOM
286
CA
GLY
X
43
14.683
0.869
21.133
1.00
43.16
C

ATOM
287
C
GLY
X
43
15.080
1.917
20.118
1.00
43.16
C

ATOM
288
O
GLY
X
43
16.264
2.083
19.830
1.00
43.16
O

ATOM
289
N
ILE
X
44
14.110
2.619
19.547
1.00
38.75
N

ATOM
290
CA
ILE
X
44
14.464
3.657
18.594
1.00
38.75
C

ATOM
291
C
ILE
X
44
14.605
4.852
19.513
1.00
38.75
C

ATOM
292
O
ILE
X
44
13.842
4.997
20.461
1.00
38.75
O

ATOM
293
CB
ILE
X
44
13.354
3.887
17.476
1.00
45.89
C

ATOM
294
CG1
ILE
X
44
12.519
5.140
17.759
1.00
45.89
C

ATOM
295
CG2
ILE
X
44
12.418
2.689
17.397
1.00
45.89
C

ATOM
296
CD1
ILE
X
44
13.108
6.423
17.216
1.00
45.89
C

ATOM
297
N
LYS
X
45
15.595
5.690
19.247
1.00
63.81
N

ATOM
298
CA
LYS
X
45
15.833
6.860
20.074
1.00
63.81
C

ATOM
299
C
LYS
X
45
15.649
8.171
19.313
1.00
63.81
C

ATOM
300
O
LYS
X
45
15.740
8.207
18.090
1.00
63.81
O

ATOM
301
CB
LYS
X
45
17.248
6.785
20.656
1.00
81.98
C

ATOM
302
CG
LYS
X
45
18.299
6.215
19.698
1.00
81.98
C

ATOM
303
CD
LYS
X
45
18.194
6.820
18.292
1.00
81.98
C

ATOM
304
CE
LYS
X
45
19.449
6.584
17.460
1.00
81.98
C

ATOM
305
NZ
LYS
X
45
20.621
7.346
18.000
1.00
81.98
N

ATOM
306
N
ASN
X
46
15.384
9.239
20.055
1.00
47.32
N

ATOM
307
CA
ASN
X
46
15.201
10.576
19.493
1.00
47.32
C

ATOM
308
C
ASN
X
46
13.819
10.790
18.915
1.00
47.32
C

ATOM
309
O
ASN
X
46
13.603
11.725
18.152
1.00
47.32
O

ATOM
310
CB
ASN
X
46
16.245
10.848
18.412
1.00
45.80
C

ATOM
311
CG
ASN
X
46
17.661
10.672
18.921
1.00
45.80
C

ATOM
312
OD1
ASN
X
46
18.061
11.308
19.894
1.00
45.80
O

ATOM
313
ND2
ASN
X
46
18.428
9.807
18.262
1.00
45.80
N

ATOM
314
N
VAL
X
47
12.880
9.938
19.312
1.00
46.74
N

ATOM
315
CA
VAL
X
47
11.516
10.007
18.814
1.00
46.74
C

ATOM
316
C
VAL
X
47
10.898
11.396
18.903
1.00
46.74
C

ATOM
317
O
VAL
X
47
9.776
11.614
18.460
1.00
46.74
O

ATOM
318
CB
VAL
X
47
10.605
8.978
19.540
1.00
30.63
C

ATOM
319
CG1
VAL
X
47
11.392
7.727
19.836
1.00
30.63
C

ATOM
320
CG2
VAL
X
47
10.022
9.563
20.793
1.00
30.63
C

ATOM
321
N
ASN
X
48
11.627
12.344
19.466
1.00
52.85
N

ATOM
322
CA
ASN
X
48
11.104
13.694
19.568
1.00
52.85
C

ATOM
323
C
ASN
X
48
11.487
14.486
18.317
1.00
52.85
C

ATOM
324
O
ASN
X
48
10.726
15.340
17.852
1.00
52.85
O

ATOM
325
CB
ASN
X
48
11.641
14.364
20.837
1.00
79.29
C

ATOM
326
CG
ASN
X
48
13.153
14.347
20.911
1.00
79.29
C

ATOM
327
OD1
ASN
X
48
13.824
15.149
20.271
1.00
79.29
O

ATOM
328
ND2
ASN
X
48
13.698
13.419
21.684
1.00
79.29
N

ATOM
329
N
LEU
X
49
12.661
14.179
17.768
1.00
63.14
N

ATOM
330
CA
LEU
X
49
13.165
14.843
16.566
1.00
63.14
C

ATOM
331
C
LEU
X
49
12.485
14.274
15.317
1.00
63.14
C

ATOM
332
O
LEU
X
49
11.821
13.239
15.384
1.00
63.14
O

ATOM
333
CB
LEU
X
49
14.682
14.647
16.455
1.00
46.75
C

ATOM
334
CG
LEU
X
49
15.558
15.152
17.614
1.00
46.75
C

ATOM
335
CD1
LEU
X
49
17.014
14.803
17.361
1.00
46.75
C

ATOM
336
CD2
LEU
X
49
15.392
16.653
17.769
1.00
46.75
C

ATOM
337
N
PRO
X
50
12.636
14.944
14.159
1.00
51.87
N

ATOM
338
CA
PRO
X
50
12.004
14.431
12.945
1.00
51.87
C

ATOM
339
C
PRO
X
50
12.700
13.143
12.577
1.00
51.87
C

ATOM
340
O
PRO
X
50
13.838
12.916
12.995
1.00
51.87
O

ATOM
341
CB
PRO
X
50
12.272
15.529
11.935
1.00
58.58
C

ATOM
342
CG
PRO
X
50
13.632
15.969
12.322
1.00
58.58
C

ATOM
343
CD
PRO
X
50
13.504
16.087
13.833
1.00
58.58
C

ATOM
344
N
PRO
X
51
12.031
12.280
11.797
1.00
45.83
N

ATOM
345
CA
PRO
X
51
10.679
12.455
11.260
1.00
45.83
C

ATOM
346
C
PRO
X
51
9.569
12.239
12.278
1.00
45.83
C

ATOM
347
O
PRO
X
51
8.518
12.880
12.203
1.00
45.83
O

ATOM
348
CB
PRO
X
51
10.626
11.422
10.161
1.00
38.67
C

ATOM
349
CG
PRO
X
51
11.410
10.307
10.773
1.00
38.67
C

ATOM
350
CD
PRO
X
51
12.615
11.021
11.305
1.00
38.67
C

ATOM
351
N
TYR
X
52
9.787
11.322
13.215
1.00
34.77
N

ATOM
352
CA
TYR
X
52
8.778
11.038
14.237
1.00
34.77
C

ATOM
353
C
TYR
X
52
8.240
12.344
14.782
1.00
34.77
C

ATOM
354
O
TYR
X
52
7.027
12.546
14.834
1.00
34.77
O

ATOM
355
CB
TYR
X
52
9.380
10.208
15.367
1.00
30.14
C

ATOM
356
CG
TYR
X
52
10.006
8.933
14.872
1.00
30.14
C

ATOM
357
CD1
TYR
X
52
11.372
8.841
14.666
1.00
30.14
C

ATOM
358
CD2
TYR
X
52
9.218
7.830
14.564
1.00
30.14
C

ATOM
359
CE1
TYR
X
52
11.943
7.680
14.163
1.00
30.14
C

ATOM
360
CE2
TYR
X
52
9.773
6.669
14.059
1.00
30.14
C

ATOM
361
CZ
TYR
X
52
11.137
6.594
13.857
1.00
30.14
C

ATOM
362
OH
TYR
X
52
11.682
5.434
13.332
1.00
30.14
O

ATOM
363
N
SER
X
53
9.161
13.221
15.186
1.00
38.91
N

ATOM
364
CA
SER
X
53
8.838
14.545
15.713
1.00
38.91
C

ATOM
365
C
SER
X
53
7.830
14.542
16.846
1.00
38.91
C

ATOM
366
O
SER
X
53
6.898
15.340
16.848
1.00
38.91
O

ATOM
367
CB
SER
X
53
8.308
15.439
14.589
1.00
35.32
C

ATOM
368
OG
SER
X
53
9.279
15.627
13.573
1.00
35.32
O

ATOM
369
N
GLY
X
54
8.003
13.645
17.808
1.00
27.35
N

ATOM
370
CA
GLY
X
54
7.078
13.600
18.927
1.00
27.35
C

ATOM
371
C
GLY
X
54
5.672
13.104
18.646
1.00
27.35
C

ATOM
372
O
GLY
X
54
4.788
13.235
19.486
1.00
27.35
O

ATOM
373
N
MET
X
55
5.429
12.537
17.476
1.00
62.97
N

ATOM
374
CA
MET
X
55
4.094
12.028
17.216
1.00
62.97
C

ATOM
375
C
MET
X
55
4.096
10.533
17.522
1.00
62.97
C

ATOM
376
O
MET
X
55
3.645
9.716
16.716
1.00
62.97
O

ATOM
377
CB
MET
X
55
3.698
12.291
15.768
1.00
39.34
C

ATOM
378
CG
MET
X
55
3.443
13.765
15.437
1.00
39.34
C

ATOM
379
SD
MET
X
55
2.351
14.625
16.593
1.00
39.34
S

ATOM
380
CE
MET
X
55
0.775
13.923
16.287
1.00
39.34
C

ATOM
381
N
THR
X
56
4.602
10.209
18.718
1.00
19.36
N

ATOM
382
CA
THR
X
56
4.753
8.854
19.240
1.00
19.36
C

ATOM
383
C
THR
X
56
3.590
8.307
20.057
1.00
19.36
C

ATOM
384
O
THR
X
56
2.721
9.046
20.500
1.00
19.36
O

ATOM
385
CB
THR
X
56
5.980
8.809
20.114
1.00
52.97
C

ATOM
386
OG1
THR
X
56
7.124
9.135
19.324
1.00
52.97
O

ATOM
387
CG2
THR
X
56
6.149
7.449
20.725
1.00
52.97
C

ATOM
388
N
ALA
X
57
3.564
6.995
20.243
1.00
21.43
N

ATOM
389
CA
ALA
X
57
2.519
6.403
21.053
1.00
21.43
C

ATOM
390
C
ALA
X
57
2.769
4.972
21.507
1.00
21.43
C

ATOM
391
O
ALA
X
57
3.665
4.288
21.017
1.00
21.43
O

ATOM
392
CB
ALA
X
57
1.205
6.501
20.354
1.00
1.67
C

ATOM
393
N
CYS
X
58
1.954
4.551
22.468
1.00
41.38
N

ATOM
394
CA
CYS
X
58
2.023
3.228
23.067
1.00
41.38
C

ATOM
395
C
CYS
X
58
0.718
2.501
22.859
1.00
41.38
C

ATOM
396
O
CYS
X
58
−0.361
3.097
22.887
1.00
41.38
O

ATOM
397
CB
CYS
X
58
2.255
3.318
24.585
1.00
51.16
C

ATOM
398
SG
CYS
X
58
3.890
3.849
25.146
1.00
51.16
S

ATOM
399
N
GLY
X
59
0.821
1.195
22.679
1.00
35.87
N

ATOM
400
CA
GLY
X
59
−0.367
0.397
22.503
1.00
35.87
C

ATOM
401
C
GLY
X
59
−0.130
−0.927
23.172
1.00
35.87
C

ATOM
402
O
GLY
X
59
1.021
−1.332
23.339
1.00
35.87
O

ATOM
403
N
ASN
X
60
−1.220
−1.587
23.554
1.00
34.54
N

ATOM
404
CA
ASN
X
60
−1.160
−2.882
24.199
1.00
34.54
C

ATOM
405
C
ASN
X
60
−0.713
−4.004
23.255
1.00
34.54
C

ATOM
406
O
ASN
X
60
−0.140
−3.750
22.194
1.00
34.54
O

ATOM
407
CB
ASN
X
60
−2.515
−3.225
24.839
1.00
40.64
C

ATOM
408
CG
ASN
X
60
−3.697
−2.642
24.083
1.00
40.64
C

ATOM
409
OD1
ASN
X
60
−3.534
−1.992
23.055
1.00
40.64
O

ATOM
410
ND2
ASN
X
60
−4.901
−2.872
24.598
1.00
40.64
N

ATOM
411
N
VAL
X
61
−0.963
−5.243
23.666
1.00
40.02
N

ATOM
412
CA
VAL
X
61
−0.580
−6.436
22.908
1.00
40.02
C

ATOM
413
C
VAL
X
61
−0.985
−6.397
21.435
1.00
40.02
C

ATOM
414
O
VAL
X
61
−0.185
−6.705
20.550
1.00
40.02
O

ATOM
415
CB
VAL
X
61
−1.204
−7.718
23.531
1.00
36.95
C

ATOM
416
CG1
VAL
X
61
−0.313
−8.915
23.256
1.00
36.95
C

ATOM
417
CG2
VAL
X
61
−1.438
−7.521
25.021
1.00
36.95
C

ATOM
418
N
PRO
X
62
−2.242
−6.025
21.161
1.00
33.00
N

ATOM
419
CA
PRO
X
62
−2.803
−5.935
19.814
1.00
33.00
C

ATOM
420
C
PRO
X
62
−1.966
−5.020
18.935
1.00
33.00
C

ATOM
421
O
PRO
X
62
−1.440
−5.418
17.894
1.00
33.00
O

ATOM
422
CB
PRO
X
62
−4.178
−5.352
20.066
1.00
21.95
C

ATOM
423
CG
PRO
X
62
−4.502
−5.810
21.448
1.00
21.95
C

ATOM
424
CD
PRO
X
62
−3.225
−5.573
22.155
1.00
21.95
C

ATOM
425
N
ILE
X
63
−1.843
−3.779
19.380
1.00
23.78
N

ATOM
426
CA
ILE
X
63
−1.092
−2.786
18.653
1.00
23.78
C

ATOM
427
C
ILE
X
63
0.388
−3.074
18.680
1.00
23.78
C

ATOM
428
O
ILE
X
63
0.994
−3.274
17.631
1.00
23.78
O

ATOM
429
CB
ILE
X
63
−1.355
−1.416
19.242
1.00
30.12
C

ATOM
430
CG1
ILE
X
63
−2.850
−1.134
19.175
1.00
30.12
C

ATOM
431
CG2
ILE
X
63
−0.570
−0.371
18.503
1.00
30.12
C

ATOM
432
CD1
ILE
X
63
−3.257
0.102
19.901
1.00
30.12
C

ATOM
433
N
PHE
X
64
0.953
−3.121
19.888
1.00
27.84
N

ATOM
434
CA
PHE
X
64
2.382
−3.352
20.101
1.00
27.84
C

ATOM
435
C
PHE
X
64
2.930
−4.704
19.654
1.00
27.84
C

ATOM
436
O
PHE
X
64
3.998
−4.764
19.053
1.00
27.84
O

ATOM
437
CB
PHE
X
64
2.738
−3.120
21.573
1.00
27.48
C

ATOM
438
CG
PHE
X
64
4.215
−3.101
21.831
1.00
27.48
C

ATOM
439
CD1
PHE
X
64
5.012
−2.124
21.266
1.00
27.48
C

ATOM
440
CD2
PHE
X
64
4.825
−4.122
22.549
1.00
27.48
C

ATOM
441
CE1
PHE
X
64
6.391
−2.164
21.397
1.00
27.48
C

ATOM
442
CE2
PHE
X
64
6.211
−4.169
22.684
1.00
27.48
C

ATOM
443
CZ
PHE
X
64
6.990
−3.190
22.104
1.00
27.48
C

ATOM
444
N
LYS
X
65
2.210
−5.784
19.947
1.00
32.57
N

ATOM
445
CA
LYS
X
65
2.639
−7.130
19.555
1.00
32.57
C

ATOM
446
C
LYS
X
65
4.087
−7.447
19.926
1.00
32.57
C

ATOM
447
O
LYS
X
65
4.945
−7.570
19.054
1.00
32.57
O

ATOM
448
CB
LYS
X
65
2.439
−7.330
18.048
1.00
29.02
C

ATOM
449
CG
LYS
X
65
0.975
−7.456
17.637
1.00
29.02
C

ATOM
450
CD
LYS
X
65
0.784
−7.344
16.120
1.00
29.02
C

ATOM
451
CE
LYS
X
65
0.914
−5.904
15.625
1.00
29.02
C

ATOM
452
NZ
LYS
X
65
1.197
−5.823
14.158
1.00
29.02
N

ATOM
453
N
ASP
X
66
4.335
−7.586
21.226
1.00
29.13
N

ATOM
454
CA
ASP
X
66
5.656
−7.896
21.777
1.00
29.13
C

ATOM
455
C
ASP
X
66
6.847
−7.411
20.965
1.00
29.13
C

ATOM
456
O
ASP
X
66
7.877
−8.097
20.894
1.00
29.13
O

ATOM
457
CB
ASP
X
66
5.816
−9.401
22.020
1.00
31.30
C

ATOM
458
CG
ASP
X
66
4.797
−9.953
23.008
1.00
31.30
C

ATOM
459
OD1
ASP
X
66
4.157
−9.159
23.740
1.00
31.30
O

ATOM
460
OD2
ASP
X
66
4.646
−11.195
23.049
1.00
31.30
O

ATOM
461
N
GLY
X
67
6.698
−6.239
20.351
1.00
28.31
N

ATOM
462
CA
GLY
X
67
7.780
−5.652
19.583
1.00
28.31
C

ATOM
463
C
GLY
X
67
7.749
−5.839
18.088
1.00
28.31
C

ATOM
464
O
GLY
X
67
8.655
−5.394
17.382
1.00
28.31
O

ATOM
465
N
LYS
X
68
6.717
−6.496
17.587
1.00
49.24
N

ATOM
466
CA
LYS
X
68
6.623
−6.727
16.155
1.00
49.24
C

ATOM
467
C
LYS
X
68
5.762
−5.654
15.506
1.00
49.24
C

ATOM
468
O
LYS
X
68
5.610
−5.613
14.290
1.00
49.24
O

ATOM
469
CB
LYS
X
68
6.076
−8.135
15.902
1.00
87.29
C

ATOM
470
CG
LYS
X
68
7.063
−9.218
16.328
1.00
87.29
C

ATOM
471
CD
LYS
X
68
6.391
−10.408
16.990
1.00
87.29
C

ATOM
472
CE
LYS
X
68
5.658
−11.287
15.992
1.00
87.29
C

ATOM
473
NZ
LYS
X
68
5.011
−12.461
16.659
1.00
87.29
N

ATOM
474
N
GLY
X
69
5.214
−4.780
16.344
1.00
37.58
N

ATOM
475
CA
GLY
X
69
4.394
−3.685
15.866
1.00
37.58
C

ATOM
476
C
GLY
X
69
5.222
−2.410
15.814
1.00
37.58
C

ATOM
477
O
GLY
X
69
4.752
−1.364
15.384
1.00
37.58
O

ATOM
478
N
CYS
X
70
6.461
−2.498
16.275
1.00
19.54
N

ATOM
479
CA
CYS
X
70
7.349
−1.359
16.269
1.00
19.54
C

ATOM
480
C
CYS
X
70
7.524
−0.878
14.859
1.00
19.54
C

ATOM
481
O
CYS
X
70
8.007
−1.632
14.023
1.00
19.54
O

ATOM
482
CB
CYS
X
70
8.712
−1.737
16.827
1.00
42.07
C

ATOM
483
SG
CYS
X
70
8.705
−1.978
18.617
1.00
42.07
S

ATOM
484
N
GLY
X
71
7.161
0.379
14.612
1.00
32.49
N

ATOM
485
CA
GLY
X
71
7.274
0.952
13.284
1.00
32.49
C

ATOM
486
C
GLY
X
71
5.906
1.277
12.703
1.00
32.49
C

ATOM
487
O
GLY
X
71
5.750
2.235
11.960
1.00
32.49
O

ATOM
488
N
SER
X
72
4.907
0.471
13.038
1.00
24.93
N

ATOM
489
CA
SER
X
72
3.556
0.691
12.553
1.00
24.93
C

ATOM
490
C
SER
X
72
3.149
2.138
12.680
1.00
24.93
C

ATOM
491
O
SER
X
72
3.662
2.873
13.511
1.00
24.93
O

ATOM
492
CB
SER
X
72
2.532
−0.115
13.360
1.00
24.64
C

ATOM
493
OG
SER
X
72
2.663
−1.496
13.172
1.00
24.64
O

ATOM
494
N
CYS
X
73
2.175
2.510
11.866
1.00
26.85
N

ATOM
495
CA
CYS
X
73
1.600
3.831
11.871
1.00
26.85
C

ATOM
496
C
CYS
X
73
0.120
3.632
12.014
1.00
26.85
C

ATOM
497
O
CYS
X
73
−0.454
2.716
11.449
1.00
26.85
O

ATOM
498
CB
CYS
X
73
1.909
4.530
10.567
1.00
30.66
C

ATOM
499
SG
CYS
X
73
3.687
4.560
10.483
1.00
30.66
S

ATOM
500
N
TYR
X
74
−0.498
4.477
12.806
1.00
24.09
N

ATOM
501
CA
TYR
X
74
−1.919
4.402
12.991
1.00
24.09
C

ATOM
502
C
TYR
X
74
−2.373
5.830
12.839
1.00
24.09
C

ATOM
503
O
TYR
X
74
−1.559
6.747
12.801
1.00
24.09
O

ATOM
504
CB
TYR
X
74
−2.266
3.856
14.374
1.00
30.54
C

ATOM
505
CG
TYR
X
74
−2.137
2.350
14.480
1.00
30.54
C

ATOM
506
CD1
TYR
X
74
−0.901
1.732
14.351
1.00
30.54
C

ATOM
507
CD2
TYR
X
74
−3.252
1.551
14.716
1.00
30.54
C

ATOM
508
CE1
TYR
X
74
−0.768
0.367
14.452
1.00
30.54
C

ATOM
509
CE2
TYR
X
74
−3.134
0.180
14.823
1.00
30.54
C

ATOM
510
CZ
TYR
X
74
−1.882
−0.406
14.690
1.00
30.54
C

ATOM
511
OH
TYR
X
74
−1.733
−1.768
14.818
1.00
30.54
O

ATOM
512
N
GLU
X
75
−3.674
6.011
12.735
1.00
34.37
N

ATOM
513
CA
GLU
X
75
−4.238
7.319
12.548
1.00
34.37
C

ATOM
514
C
GLU
X
75
−5.313
7.346
13.599
1.00
34.37
C

ATOM
515
O
GLU
X
75
−6.179
6.473
13.629
1.00
34.37
O

ATOM
516
CB
GLU
X
75
−4.840
7.400
11.135
1.00
39.78
C

ATOM
517
CG
GLU
X
75
−5.083
8.786
10.561
1.00
39.78
C

ATOM
518
CD
GLU
X
75
−5.525
8.737
9.099
1.00
39.78
C

ATOM
519
OE1
GLU
X
75
−6.625
8.219
8.806
1.00
39.78
O

ATOM
520
OE2
GLU
X
75
−4.763
9.214
8.238
1.00
39.78
O

ATOM
521
N
VAL
X
76
−5.250
8.323
14.488
1.00
31.76
N

ATOM
522
CA
VAL
X
76
−6.253
8.409
15.523
1.00
31.76
C

ATOM
523
C
VAL
X
76
−6.889
9.788
15.458
1.00
31.76
C

ATOM
524
O
VAL
X
76
−6.260
10.757
15.034
1.00
31.76
O

ATOM
525
CB
VAL
X
76
−5.627
8.134
16.907
1.00
39.03
C

ATOM
526
CG1
VAL
X
76
−4.505
9.104
17.168
1.00
39.03
C

ATOM
527
CG2
VAL
X
76
−6.696
8.206
17.985
1.00
39.03
C

ATOM
528
N
ARG
X
77
−8.150
9.865
15.857
1.00
45.60
N

ATOM
529
CA
ARG
X
77
−8.876
11.118
15.814
1.00
45.60
C

ATOM
530
C
ARG
X
77
−9.867
11.163
16.958
1.00
45.60
C

ATOM
531
O
ARG
X
77
−10.331
10.130
17.428
1.00
45.60
O

ATOM
532
CB
ARG
X
77
−9.633
11.236
14.490
1.00
74.88
C

ATOM
533
CG
ARG
X
77
−10.919
10.410
14.437
1.00
74.88
C

ATOM
534
CD
ARG
X
77
−11.539
10.418
13.051
1.00
74.88
C

ATOM
535
NE
ARG
X
77
−12.867
9.810
13.026
1.00
74.88
N

ATOM
536
CZ
ARG
X
77
−13.539
9.531
11.912
1.00
74.88
C

ATOM
537
NH1
ARG
X
77
−13.006
9.800
10.726
1.00
74.88
N

ATOM
538
NH2
ARG
X
77
−14.751
8.994
11.981
1.00
74.88
N

ATOM
539
N
CYS
X
78
−10.195
12.366
17.403
1.00
50.61
N

ATOM
540
CA
CYS
X
78
−11.142
12.510
18.486
1.00
50.61
C

ATOM
541
C
CYS
X
78
−12.384
13.247
18.037
1.00
50.61
C

ATOM
542
O
CYS
X
78
−12.304
14.219
17.294
1.00
50.61
O

ATOM
543
CB
CYS
X
78
−10.529
13.272
19.655
1.00
84.27
C

ATOM
544
SG
CYS
X
78
−11.853
13.899
20.726
1.00
84.27
S

ATOM
545
N
LYS
X
79
−13.536
12.788
18.501
1.00
55.20
N

ATOM
546
CA
LYS
X
79
−14.784
13.435
18.159
1.00
55.20
C

ATOM
547
C
LYS
X
79
−15.804
13.199
19.262
1.00
55.20
C

ATOM
548
O
LYS
X
79
−17.007
13.194
19.018
1.00
55.20
O

ATOM
549
CB
LYS
X
79
−15.310
12.906
16.822
1.00
64.46
C

ATOM
550
CG
LYS
X
79
−16.551
13.644
16.321
1.00
64.46
C

ATOM
551
CD
LYS
X
79
−16.319
15.161
16.245
1.00
64.46
C

ATOM
552
CE
LYS
X
79
−17.621
15.940
16.100
1.00
64.46
C

ATOM
553
NZ
LYS
X
79
−18.501
15.773
17.291
1.00
64.46
N

ATOM
554
N
GLU
X
80
−15.320
13.005
20.485
1.00
69.81
N

ATOM
555
CA
GLU
X
80
−16.216
12.773
21.610
1.00
69.81
C

ATOM
556
C
GLU
X
80
−16.364
14.053
22.413
1.00
69.81
C

ATOM
557
O
GLU
X
80
−17.442
14.643
22.482
1.00
69.81
O

ATOM
558
CB
GLU
X
80
−15.674
11.667
22.517
1.00
99.00
C

ATOM
559
CG
GLU
X
80
−16.758
10.860
23.228
1.00
99.00
C

ATOM
560
CD
GLU
X
80
−17.868
11.725
23.814
1.00
99.00
C

ATOM
561
OE1
GLU
X
80
−18.648
12.320
23.036
1.00
99.00
O

ATOM
562
OE2
GLU
X
80
−17.963
11.812
25.057
1.00
99.00
O

ATOM
563
N
LYS
X
81
−15.269
14.483
23.024
1.00
99.00
N

ATOM
564
CA
LYS
X
81
−15.290
15.699
23.814
1.00
99.00
C

ATOM
565
C
LYS
X
81
−14.927
16.869
22.907
1.00
99.00
C

ATOM
566
O
LYS
X
81
−14.218
16.693
21.914
1.00
99.00
O

ATOM
567
CB
LYS
X
81
−14.308
15.575
24.981
1.00
79.10
C

ATOM
568
CG
LYS
X
81
−14.499
14.279
25.763
1.00
79.10
C

ATOM
569
CD
LYS
X
81
−13.766
14.282
27.089
1.00
79.10
C

ATOM
570
CE
LYS
X
81
−14.398
15.257
28.083
1.00
79.10
C

ATOM
571
NZ
LYS
X
81
−15.785
14.877
28.493
1.00
79.10
N

ATOM
572
N
PRO
X
82
−15.426
18.076
23.228
1.00
70.36
N

ATOM
573
CA
PRO
X
82
−15.166
19.289
22.448
1.00
70.36
C

ATOM
574
C
PRO
X
82
−13.689
19.488
22.169
1.00
70.36
C

ATOM
575
O
PRO
X
82
−12.902
18.552
22.263
1.00
70.36
O

ATOM
576
CB
PRO
X
82
−15.736
20.391
23.331
1.00
84.04
C

ATOM
577
CG
PRO
X
82
−16.875
19.705
24.012
1.00
84.04
C

ATOM
578
CD
PRO
X
82
−16.256
18.388
24.405
1.00
84.04
C

ATOM
579
N
GLU
X
83
−13.321
20.717
21.836
1.00
55.17
N

ATOM
580
CA
GLU
X
83
−11.933
21.063
21.534
1.00
55.17
C

ATOM
581
C
GLU
X
83
−11.203
19.975
20.756
1.00
55.17
C

ATOM
582
O
GLU
X
83
−9.971
19.939
20.723
1.00
55.17
O

ATOM
583
CB
GLU
X
83
−11.150
21.397
22.821
1.00
75.84
C

ATOM
584
CG
GLU
X
83
−10.952
20.259
23.812
1.00
75.84
C

ATOM
585
CD
GLU
X
83
−11.921
20.318
24.981
1.00
75.84
C

ATOM
586
OE1
GLU
X
83
−13.117
20.012
24.794
1.00
75.84
O

ATOM
587
OE2
GLU
X
83
−11.483
20.680
26.093
1.00
75.84
O

ATOM
588
N
CYS
X
84
−11.975
19.097
20.122
1.00
57.58
N

ATOM
589
CA
CYS
X
84
−11.407
18.014
19.345
1.00
57.58
C

ATOM
590
C
CYS
X
84
−11.399
18.340
17.881
1.00
57.58
C

ATOM
591
O
CYS
X
84
−12.441
18.397
17.244
1.00
57.58
O

ATOM
592
CB
CYS
X
84
−12.165
16.718
19.610
1.00
84.27
C

ATOM
593
SG
CYS
X
84
−11.401
15.849
21.010
1.00
84.27
S

ATOM
594
N
SER
X
85
−10.197
18.570
17.368
1.00
50.63
N

ATOM
595
CA
SER
X
85
−9.987
18.911
15.972
1.00
50.63
C

ATOM
596
C
SER
X
85
−10.933
18.155
15.041
1.00
50.63
C

ATOM
597
O
SER
X
85
−11.336
18.669
13.989
1.00
50.63
O

ATOM
598
CB
SER
X
85
−8.532
18.620
15.577
1.00
76.77
C

ATOM
599
OG
SER
X
85
−8.219
17.239
15.698
1.00
76.77
O

ATOM
600
N
GLY
X
86
−11.300
16.942
15.441
1.00
48.12
N

ATOM
601
CA
GLY
X
86
−12.168
16.132
14.614
1.00
48.12
C

ATOM
602
C
GLY
X
86
−11.291
15.422
13.600
1.00
48.12
C

ATOM
603
O
GLY
X
86
−11.585
14.310
13.168
1.00
48.12
O

ATOM
604
N
ASN
X
87
−10.196
16.072
13.221
1.00
69.99
N

ATOM
605
CA
ASN
X
87
−9.266
15.502
12.261
1.00
69.99
C

ATOM
606
C
ASN
X
87
−8.284
14.559
12.941
1.00
69.99
C

ATOM
607
O
ASN
X
87
−7.938
14.740
14.110
1.00
69.99
O

ATOM
608
CB
ASN
X
87
−8.515
16.616
11.529
1.00
95.79
C

ATOM
609
CG
ASN
X
87
−9.240
17.080
10.283
1.00
95.79
C

ATOM
610
OD1
ASN
X
87
−10.441
17.352
10.312
1.00
95.79
O

ATOM
611
ND2
ASN
X
87
−8.511
17.174
9.177
1.00
95.79
N

ATOM
612
N
PRO
X
88
−7.818
13.537
12.205
1.00
50.49
N

ATOM
613
CA
PRO
X
88
−6.878
12.544
12.711
1.00
50.49
C

ATOM
614
C
PRO
X
88
−5.429
12.998
12.684
1.00
50.49
C

ATOM
615
O
PRO
X
88
−5.057
13.908
11.949
1.00
50.49
O

ATOM
616
CB
PRO
X
88
−7.116
11.366
11.780
1.00
54.49
C

ATOM
617
CG
PRO
X
88
−7.288
12.059
10.460
1.00
54.49
C

ATOM
618
CD
PRO
X
88
−8.223
13.202
10.824
1.00
54.49
C

ATOM
619
N
VAL
X
89
−4.616
12.340
13.499
1.00
61.42
N

ATOM
620
CA
VAL
X
89
−3.192
12.621
13.575
1.00
61.42
C

ATOM
621
C
VAL
X
89
−2.520
11.295
13.260
1.00
61.42
C

ATOM
622
O
VAL
X
89
−3.058
10.242
13.581
1.00
61.42
O

ATOM
623
CB
VAL
X
89
−2.769
13.088
15.003
1.00
47.66
C

ATOM
624
CG1
VAL
X
89
−2.241
14.496
14.949
1.00
47.66
C

ATOM
625
CG2
VAL
X
89
−3.948
13.029
15.957
1.00
47.66
C

ATOM
626
N
THR
X
90
−1.369
11.333
12.609
1.00
52.85
N

ATOM
627
CA
THR
X
90
−0.672
10.096
12.297
1.00
52.85
C

ATOM
628
C
THR
X
90
0.392
9.825
13.353
1.00
52.85
C

ATOM
629
O
THR
X
90
1.376
10.557
13.469
1.00
52.85
O

ATOM
630
CB
THR
X
90
−0.007
10.130
10.889
1.00
49.85
C

ATOM
631
OG1
THR
X
90
−1.016
10.024
9.878
1.00
49.85
O

ATOM
632
CG2
THR
X
90
0.959
8.974
10.723
1.00
49.85
C

ATOM
633
N
VAL
X
91
0.178
8.756
14.112
1.00
51.89
N

ATOM
634
CA
VAL
X
91
1.076
8.341
15.179
1.00
51.89
C

ATOM
635
C
VAL
X
91
1.984
7.167
14.796
1.00
51.89
C

ATOM
636
O
VAL
X
91
1.651
6.373
13.917
1.00
51.89
O

ATOM
637
CB
VAL
X
91
0.258
7.922
16.408
1.00
36.09
C

ATOM
638
CG1
VAL
X
91
1.098
7.103
17.321
1.00
36.09
C

ATOM
639
CG2
VAL
X
91
−0.265
9.136
17.126
1.00
36.09
C

ATOM
640
N
TYR
X
92
3.143
7.079
15.446
1.00
54.85
N

ATOM
641
CA
TYR
X
92
4.063
5.967
15.228
1.00
54.85
C

ATOM
642
C
TYR
X
92
4.060
5.165
16.518
1.00
54.85
C

ATOM
643
O
TYR
X
92
3.809
5.712
17.589
1.00
54.85
O

ATOM
644
CB
TYR
X
92
5.483
6.441
14.949
1.00
51.94
C

ATOM
645
CG
TYR
X
92
5.677
7.072
13.592
1.00
51.94
C

ATOM
646
CD1
TYR
X
92
5.221
8.367
13.332
1.00
51.94
C

ATOM
647
CD2
TYR
X
92
6.344
6.387
12.569
1.00
51.94
C

ATOM
648
CE1
TYR
X
92
5.429
8.970
12.088
1.00
51.94
C

ATOM
649
CE2
TYR
X
92
6.554
6.981
11.327
1.00
51.94
C

ATOM
650
CZ
TYR
X
92
6.094
8.274
11.100
1.00
51.94
C

ATOM
651
OH
TYR
X
92
6.315
8.879
9.892
1.00
51.94
O

ATOM
652
N
ILE
X
93
4.310
3.866
16.422
1.00
35.81
N

ATOM
653
CA
ILE
X
93
4.339
3.034
17.616
1.00
35.81
C

ATOM
654
C
ILE
X
93
5.795
2.737
17.945
1.00
35.81
C

ATOM
655
O
ILE
X
93
6.447
1.924
17.295
1.00
35.81
O

ATOM
656
CB
ILE
X
93
3.542
1.732
17.407
1.00
25.05
C

ATOM
657
CG1
ILE
X
93
2.067
2.069
17.238
1.00
25.05
C

ATOM
658
CG2
ILE
X
93
3.705
0.815
18.589
1.00
25.05
C

ATOM
659
CD1
ILE
X
93
1.550
3.019
18.292
1.00
25.05
C

ATOM
660
N
THR
X
94
6.304
3.425
18.955
1.00
44.25
N

ATOM
661
CA
THR
X
94
7.689
3.260
19.337
1.00
44.25
C

ATOM
662
C
THR
X
94
7.818
2.816
20.773
1.00
44.25
C

ATOM
663
O
THR
X
94
8.930
2.680
21.290
1.00
44.25
O

ATOM
664
CB
THR
X
94
8.450
4.569
19.182
1.00
31.85
C

ATOM
665
OG1
THR
X
94
7.878
5.545
20.055
1.00
31.85
O

ATOM
666
CG2
THR
X
94
8.365
5.067
17.764
1.00
31.85
C

ATOM
667
N
ASP
X
95
6.687
2.594
21.423
1.00
41.33
N

ATOM
668
CA
ASP
X
95
6.734
2.168
22.804
1.00
41.33
C

ATOM
669
C
ASP
X
95
5.582
1.248
23.159
1.00
41.33
C

ATOM
670
O
ASP
X
95
4.811
0.860
22.287
1.00
41.33
O

ATOM
671
CB
ASP
X
95
6.749
3.385
23.719
1.00
46.56
C

ATOM
672
CG
ASP
X
95
7.241
3.056
25.107
1.00
46.56
C

ATOM
673
OD1
ASP
X
95
8.263
2.344
25.219
1.00
46.56
O

ATOM
674
OD2
ASP
X
95
6.609
3.520
26.074
1.00
46.56
O

ATOM
675
N
MET
X
96
5.459
0.901
24.438
1.00
38.95
N

ATOM
676
CA
MET
X
96
4.403
−0.006
24.854
1.00
38.95
C

ATOM
677
C
MET
X
96
3.818
0.242
26.226
1.00
38.95
C

ATOM
678
O
MET
X
96
4.406
0.917
27.052
1.00
38.95
O

ATOM
679
CB
MET
X
96
4.912
−1.445
24.803
1.00
48.57
C

ATOM
680
CG
MET
X
96
6.080
−1.708
25.727
1.00
48.57
C

ATOM
681
SD
MET
X
96
5.962
−3.305
26.529
1.00
48.57
S

ATOM
682
CE
MET
X
96
4.657
−2.957
27.709
1.00
48.57
C

ATOM
683
N
ASN
X
97
2.644
−0.331
26.443
1.00
29.42
N

ATOM
684
CA
ASN
X
97
1.928
−0.253
27.704
1.00
29.42
C

ATOM
685
C
ASN
X
97
0.850
−1.350
27.692
1.00
29.42
C

ATOM
686
O
ASN
X
97
−0.141
−1.265
26.963
1.00
29.42
O

ATOM
687
CB
ASN
X
97
1.295
1.119
27.886
1.00
38.27
C

ATOM
688
CG
ASN
X
97
0.453
1.204
29.152
1.00
38.27
C

ATOM
689
OD1
ASN
X
97
−0.171
2.229
29.431
1.00
38.27
O

ATOM
690
ND2
ASN
X
97
0.429
0.122
29.923
1.00
38.27
N

ATOM
691
N
TYR
X
98
1.048
−2.383
28.506
1.00
52.46
N

ATOM
692
CA
TYR
X
98
0.112
−3.496
28.541
1.00
52.46
C

ATOM
693
C
TYR
X
98
−0.962
−3.480
29.620
1.00
52.46
C

ATOM
694
O
TYR
X
98
−1.615
−4.491
29.844
1.00
52.46
O

ATOM
695
CB
TYR
X
98
0.891
−4.809
28.610
1.00
44.42
C

ATOM
696
CG
TYR
X
98
1.520
−5.214
27.289
1.00
44.42
C

ATOM
697
CD1
TYR
X
98
2.016
−4.258
26.400
1.00
44.42
C

ATOM
698
CD2
TYR
X
98
1.627
−6.556
26.929
1.00
44.42
C

ATOM
699
CE1
TYR
X
98
2.603
−4.634
25.180
1.00
44.42
C

ATOM
700
CE2
TYR
X
98
2.213
−6.943
25.718
1.00
44.42
C

ATOM
701
CZ
TYR
X
98
2.698
−5.978
24.850
1.00
44.42
C

ATOM
702
OH
TYR
X
98
3.286
−6.362
23.667
1.00
44.42
O

ATOM
703
N
GLU
X
99
−1.168
−2.345
30.277
1.00
31.89
N

ATOM
704
CA
GLU
X
99
−2.189
−2.275
31.316
1.00
31.89
C

ATOM
705
C
GLU
X
99
−3.540
−2.672
30.748
1.00
31.89
C

ATOM
706
O
GLU
X
99
−4.013
−2.100
29.769
1.00
31.89
O

ATOM
707
CB
GLU
X
99
−2.264
−0.864
31.902
1.00
68.33
C

ATOM
708
CG
GLU
X
99
−1.155
−0.542
32.893
1.00
68.33
C

ATOM
709
CD
GLU
X
99
−1.185
0.905
33.349
1.00
68.33
C

ATOM
710
OE1
GLU
X
99
−2.288
1.406
33.659
1.00
68.33
O

ATOM
711
OE2
GLU
X
99
−0.107
1.538
33.403
1.00
68.33
O

ATOM
712
N
PRO
X
100
−4.189
−3.666
31.357
1.00
65.90
N

ATOM
713
CA
PRO
X
100
−5.495
−4.080
30.836
1.00
65.90
C

ATOM
714
C
PRO
X
100
−6.634
−3.103
31.124
1.00
65.90
C

ATOM
715
O
PRO
X
100
−7.674
−3.519
31.621
1.00
65.90
O

ATOM
716
CB
PRO
X
100
−5.725
−5.432
31.517
1.00
77.94
C

ATOM
717
CG
PRO
X
100
−4.319
−5.918
31.829
1.00
77.94
C

ATOM
718
CD
PRO
X
100
−3.660
−4.659
32.307
1.00
77.94
C

ATOM
719
N
ILE
X
101
−6.465
−1.819
30.812
1.00
48.28
N

ATOM
720
CA
ILE
X
101
−7.542
−0.859
31.085
1.00
48.28
C

ATOM
721
C
ILE
X
101
−8.640
−0.869
30.036
1.00
48.28
C

ATOM
722
O
ILE
X
101
−9.745
−0.406
30.302
1.00
48.28
O

ATOM
723
CB
ILE
X
101
−7.046
0.587
31.172
1.00
55.27
C

ATOM
724
CG1
ILE
X
101
−6.564
1.050
29.800
1.00
55.27
C

ATOM
725
CG2
ILE
X
101
−5.954
0.700
32.210
1.00
55.27
C

ATOM
726
CD1
ILE
X
101
−6.231
2.515
29.745
1.00
55.27
C

ATOM
727
N
ALA
X
102
−8.334
−1.372
28.840
1.00
49.68
N

ATOM
728
CA
ALA
X
102
−9.327
−1.442
27.771
1.00
49.68
C

ATOM
729
C
ALA
X
102
−8.861
−2.352
26.651
1.00
49.68
C

ATOM
730
O
ALA
X
102
−7.669
−2.602
26.509
1.00
49.68
O

ATOM
731
CB
ALA
X
102
−9.604
−0.058
27.224
1.00
50.56
C

ATOM
732
N
PRO
X
103
−9.804
−2.862
25.840
1.00
33.37
N

ATOM
733
CA
PRO
X
103
−9.506
−3.750
24.716
1.00
33.37
C

ATOM
734
C
PRO
X
103
−8.323
−3.215
23.928
1.00
33.37
C

ATOM
735
O
PRO
X
103
−7.287
−3.865
23.839
1.00
33.37
O

ATOM
736
CB
PRO
X
103
−10.804
−3.727
23.922
1.00
31.39
C

ATOM
737
CG
PRO
X
103
−11.829
−3.671
24.995
1.00
31.39
C

ATOM
738
CD
PRO
X
103
−11.256
−2.619
25.931
1.00
31.39
C

ATOM
739
N
TYR
X
104
−8.482
−2.029
23.345
1.00
49.21
N

ATOM
740
CA
TYR
X
104
−7.392
−1.393
22.599
1.00
49.21
C

ATOM
741
C
TYR
X
104
−7.017
−0.122
23.343
1.00
49.21
C

ATOM
742
O
TYR
X
104
−7.884
0.685
23.675
1.00
49.21
O

ATOM
743
CB
TYR
X
104
−7.806
−1.035
21.168
1.00
45.54
C

ATOM
744
CG
TYR
X
104
−8.016
−2.213
20.242
1.00
45.54
C

ATOM
745
CD1
TYR
X
104
−9.302
−2.643
19.911
1.00
45.54
C

ATOM
746
CD2
TYR
X
104
−6.934
−2.864
19.659
1.00
45.54
C

ATOM
747
CE1
TYR
X
104
−9.500
−3.684
19.018
1.00
45.54
C

ATOM
748
CE2
TYR
X
104
−7.120
−3.903
18.770
1.00
45.54
C

ATOM
749
CZ
TYR
X
104
−8.405
−4.306
18.450
1.00
45.54
C

ATOM
750
OH
TYR
X
104
−8.596
−5.315
17.540
1.00
45.54
O

ATOM
751
N
HIS
X
105
−5.726
0.059
23.591
1.00
29.90
N

ATOM
752
CA
HIS
X
105
−5.264
1.220
24.327
1.00
29.90
C

ATOM
753
C
HIS
X
105
−4.078
1.933
23.707
1.00
29.90
C

ATOM
754
O
HIS
X
105
−3.072
1.310
23.381
1.00
29.90
O

ATOM
755
CB
HIS
X
105
−4.915
0.794
25.748
1.00
64.97
C

ATOM
756
CG
HIS
X
105
−4.261
1.865
26.558
1.00
64.97
C

ATOM
757
ND1
HIS
X
105
−3.024
1.701
27.140
1.00
64.97
N

ATOM
758
CD2
HIS
X
105
−4.678
3.106
26.899
1.00
64.97
C

ATOM
759
CE1
HIS
X
105
−2.707
2.796
27.808
1.00
64.97
C

ATOM
760
NE2
HIS
X
105
−3.694
3.664
27.678
1.00
64.97
N

ATOM
761
N
PHE
X
106
−4.211
3.247
23.542
1.00
36.72
N

ATOM
762
CA
PHE
X
106
−3.138
4.074
22.994
1.00
36.72
C

ATOM
763
C
PHE
X
106
−2.725
4.999
24.112
1.00
36.72
C

ATOM
764
O
PHE
X
106
−3.571
5.569
24.803
1.00
36.72
O

ATOM
765
CB
PHE
X
106
−3.599
4.955
21.818
1.00
34.76
C

ATOM
766
CG
PHE
X
106
−3.737
4.230
20.515
1.00
34.76
C

ATOM
767
CD1
PHE
X
106
−4.923
3.576
20.183
1.00
34.76
C

ATOM
768
CD2
PHE
X
106
−2.676
4.194
19.613
1.00
34.76
C

ATOM
769
CE1
PHE
X
106
−5.049
2.899
18.972
1.00
34.76
C

ATOM
770
CE2
PHE
X
106
−2.794
3.517
18.397
1.00
34.76
C

ATOM
771
CZ
PHE
X
106
−3.981
2.872
18.080
1.00
34.76
C

ATOM
772
N
ASP
X
107
−1.422
5.145
24.287
1.00
25.99
N

ATOM
773
CA
ASP
X
107
−0.879
6.021
25.310
1.00
25.99
C

ATOM
774
C
ASP
X
107
−0.043
6.976
24.495
1.00
25.99
C

ATOM
775
O
ASP
X
107
1.114
6.703
24.183
1.00
25.99
O

ATOM
776
CB
ASP
X
107
0.004
5.234
26.286
1.00
56.65
C

ATOM
777
CG
ASP
X
107
−0.010
5.817
27.690
1.00
56.65
C

ATOM
778
OD1
ASP
X
107
0.816
5.389
28.517
1.00
56.65
O

ATOM
779
OD2
ASP
X
107
−0.850
6.692
27.974
1.00
56.65
O

ATOM
780
N
LEU
X
108
−0.643
8.096
24.134
1.00
46.68
N

ATOM
781
CA
LEU
X
108
0.037
9.062
23.300
1.00
46.68
C

ATOM
782
C
LEU
X
108
1.006
9.953
24.046
1.00
46.68
C

ATOM
783
O
LEU
X
108
1.015
10.012
25.270
1.00
46.68
O

ATOM
784
CB
LEU
X
108
−1.003
9.919
22.578
1.00
36.48
C

ATOM
785
CG
LEU
X
108
−2.051
9.128
21.783
1.00
36.48
C

ATOM
786
CD1
LEU
X
108
−3.179
10.040
21.345
1.00
36.48
C

ATOM
787
CD2
LEU
X
108
−1.392
8.462
20.592
1.00
36.48
C

ATOM
788
N
SER
X
109
1.844
10.630
23.283
1.00
37.04
N

ATOM
789
CA
SER
X
109
2.792
11.572
23.834
1.00
37.04
C

ATOM
790
C
SER
X
109
1.946
12.809
24.130
1.00
37.04
C

ATOM
791
O
SER
X
109
0.773
12.868
23.752
1.00
37.04
O

ATOM
792
CB
SER
X
109
3.839
11.918
22.781
1.00
35.78
C

ATOM
793
OG
SER
X
109
3.209
12.477
21.642
1.00
35.78
O

ATOM
794
N
GLY
X
110
2.528
13.797
24.801
1.00
45.00
N

ATOM
795
CA
GLY
X
110
1.775
15.006
25.086
1.00
45.00
C

ATOM
796
C
GLY
X
110
1.492
15.712
23.774
1.00
45.00
C

ATOM
797
O
GLY
X
110
0.375
16.162
23.515
1.00
45.00
O

ATOM
798
N
LYS
X
111
2.525
15.793
22.943
1.00
42.59
N

ATOM
799
CA
LYS
X
111
2.441
16.423
21.630
1.00
42.59
C

ATOM
800
C
LYS
X
111
1.277
15.820
20.850
1.00
42.59
C

ATOM
801
O
LYS
X
111
0.321
16.518
20.494
1.00
42.59
O

ATOM
802
CB
LYS
X
111
3.748
16.183
20.878
1.00
36.13
C

ATOM
803
CG
LYS
X
111
3.972
17.011
19.635
1.00
36.13
C

ATOM
804
CD
LYS
X
111
5.397
16.781
19.164
1.00
36.13
C

ATOM
805
CE
LYS
X
111
5.834
17.775
18.107
1.00
36.13
C

ATOM
806
NZ
LYS
X
111
5.108
17.577
16.820
1.00
36.13
N

ATOM
807
N
ALA
X
112
1.368
14.514
20.596
1.00
54.93
N

ATOM
808
CA
ALA
X
112
0.339
13.786
19.863
1.00
54.93
C

ATOM
809
C
ALA
X
112
−1.027
13.978
20.498
1.00
54.93
C

ATOM
810
O
ALA
X
112
−1.941
14.493
19.872
1.00
54.93
O

ATOM
811
CB
ALA
X
112
0.678
12.307
19.819
1.00
46.83
C

ATOM
812
N
PHE
X
113
−1.167
13.571
21.748
1.00
52.54
N

ATOM
813
CA
PHE
X
113
−2.448
13.701
22.422
1.00
52.54
C

ATOM
814
C
PHE
X
113
−3.049
15.087
22.222
1.00
52.54
C

ATOM
815
O
PHE
X
113
−4.263
15.229
22.051
1.00
52.54
O

ATOM
816
CB
PHE
X
113
−2.294
13.433
23.915
1.00
34.42
C

ATOM
817
CG
PHE
X
113
−3.591
13.167
24.606
1.00
34.42
C

ATOM
818
CD1
PHE
X
113
−3.941
11.876
24.969
1.00
34.42
C

ATOM
819
CD2
PHE
X
113
−4.486
14.196
24.841
1.00
34.42
C

ATOM
820
CE1
PHE
X
113
−5.167
11.607
25.552
1.00
34.42
C

ATOM
821
CE2
PHE
X
113
−5.723
13.939
25.427
1.00
34.42
C

ATOM
822
CZ
PHE
X
113
−6.067
12.637
25.783
1.00
34.42
C

ATOM
823
N
GLY
X
114
−2.194
16.107
22.254
1.00
62.96
N

ATOM
824
CA
GLY
X
114
−2.659
17.472
22.078
1.00
62.96
C

ATOM
825
C
GLY
X
114
−3.149
17.732
20.670
1.00
62.96
C

ATOM
826
O
GLY
X
114
−4.180
18.375
20.475
1.00
62.96
O

ATOM
827
N
SER
X
115
−2.404
17.220
19.693
1.00
41.75
N

ATOM
828
CA
SER
X
115
−2.734
17.373
18.282
1.00
41.75
C

ATOM
829
C
SER
X
115
−4.139
16.942
17.914
1.00
41.75
C

ATOM
830
O
SER
X
115
−4.631
17.271
16.842
1.00
41.75
O

ATOM
831
CB
SER
X
115
−1.743
16.599
17.427
1.00
31.27
C

ATOM
832
OG
SER
X
115
−0.458
17.184
17.502
1.00
31.27
O

ATOM
833
N
LEU
X
116
−4.797
16.200
18.785
1.00
36.02
N

ATOM
834
CA
LEU
X
116
−6.149
15.774
18.473
1.00
36.02
C

ATOM
835
C
LEU
X
116
−7.111
16.927
18.683
1.00
36.02
C

ATOM
836
O
LEU
X
116
−8.313
16.787
18.456
1.00
36.02
O

ATOM
837
CB
LEU
X
116
−6.563
14.606
19.369
1.00
67.90
C

ATOM
838
CG
LEU
X
116
−5.945
13.238
19.085
1.00
67.90
C

ATOM
839
CD1
LEU
X
116
−6.298
12.271
20.203
1.00
67.90
C

ATOM
840
CD2
LEU
X
116
−6.460
12.723
17.756
1.00
67.90
C

ATOM
841
N
ALA
X
117
−6.580
18.074
19.104
1.00
67.56
N

ATOM
842
CA
ALA
X
117
−7.409
19.247
19.400
1.00
67.56
C

ATOM
843
C
ALA
X
117
−7.461
20.372
18.376
1.00
67.56
C

ATOM
844
O
ALA
X
117
−6.535
20.577
17.595
1.00
67.56
O

ATOM
845
CB
ALA
X
117
−6.996
19.826
20.749
1.00
67.72
C

ATOM
846
N
LYS
X
118
−8.566
21.107
18.405
1.00
62.31
N

ATOM
847
CA
LYS
X
118
−8.756
22.246
17.520
1.00
62.31
C

ATOM
848
C
LYS
X
118
−7.646
23.219
17.901
1.00
62.31
C

ATOM
849
O
LYS
X
118
−7.308
23.349
19.076
1.00
62.31
O

ATOM
850
CB
LYS
X
118
−10.121
22.887
17.774
1.00
70.06
C

ATOM
851
CG
LYS
X
118
−11.287
21.909
17.729
1.00
70.06
C

ATOM
852
CD
LYS
X
118
−12.588
22.547
18.219
1.00
70.06
C

ATOM
853
CE
LYS
X
118
−13.162
23.554
17.225
1.00
70.06
C

ATOM
854
NZ
LYS
X
118
−13.743
22.908
16.013
1.00
70.06
N

ATOM
855
N
PRO
X
119
−7.070
23.920
16.916
1.00
70.89
N

ATOM
856
CA
PRO
X
119
−5.989
24.880
17.166
1.00
70.89
C

ATOM
857
C
PRO
X
119
−6.164
25.752
18.409
1.00
70.89
C

ATOM
858
O
PRO
X
119
−7.208
26.385
18.598
1.00
70.89
O

ATOM
859
CB
PRO
X
119
−5.958
25.697
15.881
1.00
73.89
C

ATOM
860
CG
PRO
X
119
−6.308
24.670
14.851
1.00
73.89
C

ATOM
861
CD
PRO
X
119
−7.472
23.948
15.498
1.00
73.89
C

ATOM
862
N
GLY
X
120
−5.130
25.764
19.252
1.00
72.64
N

ATOM
863
CA
GLY
X
120
−5.138
26.561
20.470
1.00
72.64
C

ATOM
864
C
GLY
X
120
−5.803
25.937
21.682
1.00
72.64
C

ATOM
865
O
GLY
X
120
−5.872
26.557
22.744
1.00
72.64
O

ATOM
866
N
LEU
X
121
−6.278
24.704
21.534
1.00
98.68
N

ATOM
867
CA
LEU
X
121
−6.955
24.012
22.625
1.00
98.68
C

ATOM
868
C
LEU
X
121
−6.145
22.839
23.185
1.00
98.68
C

ATOM
869
O
LEU
X
121
−6.619
22.100
24.047
1.00
98.68
O

ATOM
870
CB
LEU
X
121
−8.324
23.533
22.134
1.00
57.37
C

ATOM
871
CG
LEU
X
121
−9.223
24.670
21.636
1.00
57.37
C

ATOM
872
CD1
LEU
X
121
−10.441
24.133
20.912
1.00
57.37
C

ATOM
873
CD2
LEU
X
121
−9.643
25.510
22.814
1.00
57.37
C

ATOM
874
N
ASN
X
122
−4.920
22.688
22.697
1.00
66.63
N

ATOM
875
CA
ASN
X
122
−4.028
21.617
23.125
1.00
66.63
C

ATOM
876
C
ASN
X
122
−4.042
21.372
24.630
1.00
66.63
C

ATOM
877
O
ASN
X
122
−4.832
20.574
25.136
1.00
66.63
O

ATOM
878
CB
ASN
X
122
−2.600
21.935
22.689
1.00
75.00
C

ATOM
879
CG
ASN
X
122
−2.524
22.355
21.247
1.00
75.00
C

ATOM
880
OD1
ASN
X
122
−3.178
23.313
20.843
1.00
75.00
O

ATOM
881
ND2
ASN
X
122
−1.729
21.642
20.456
1.00
75.00
N

ATOM
882
N
ASP
X
123
−3.140
22.058
25.329
1.00
76.12
N

ATOM
883
CA
ASP
X
123
−2.991
21.952
26.780
1.00
76.12
C

ATOM
884
C
ASP
X
123
−4.332
21.807
27.480
1.00
76.12
C

ATOM
885
O
ASP
X
123
−4.436
21.173
28.527
1.00
76.12
O

ATOM
886
CB
ASP
X
123
−2.230
23.171
27.320
1.00
85.08
C

ATOM
887
CG
ASP
X
123
−1.873
24.177
26.226
1.00
85.08
C

ATOM
888
OD1
ASP
X
123
−0.770
24.768
26.285
1.00
85.08
O

ATOM
889
OD2
ASP
X
123
−2.701
24.389
25.313
1.00
85.08
O

ATOM
890
N
LYS
X
124
−5.362
22.394
26.894
1.00
66.79
N

ATOM
891
CA
LYS
X
124
−6.688
22.294
27.465
1.00
66.79
C

ATOM
892
C
LYS
X
124
−7.091
20.828
27.457
1.00
66.79
C

ATOM
893
O
LYS
X
124
−7.331
20.227
28.504
1.00
66.79
O

ATOM
894
CB
LYS
X
124
−7.675
23.093
26.626
1.00
84.27
C

ATOM
895
CG
LYS
X
124
−9.096
23.060
27.139
1.00
84.27
C

ATOM
896
CD
LYS
X
124
−10.013
23.849
26.232
1.00
84.27
C

ATOM
897
CE
LYS
X
124
−9.509
25.278
26.057
1.00
84.27
C

ATOM
898
NZ
LYS
X
124
−8.222
25.348
25.282
1.00
84.27
N

ATOM
899
N
ILE
X
125
−7.143
20.266
26.252
1.00
75.63
N

ATOM
900
CA
ILE
X
125
−7.529
18.876
26.019
1.00
75.63
C

ATOM
901
C
ILE
X
125
−6.622
17.872
26.746
1.00
75.63
C

ATOM
902
O
ILE
X
125
−7.072
16.790
27.126
1.00
75.63
O

ATOM
903
CB
ILE
X
125
−7.529
18.560
24.487
1.00
75.19
C

ATOM
904
CG1
ILE
X
125
−8.471
17.387
24.172
1.00
35.80
C

ATOM
905
CG2
ILE
X
125
−6.102
18.233
24.020
1.00
35.80
C

ATOM
906
CD1
ILE
X
125
−8.440
16.933
22.709
1.00
75.19
C

ATOM
907
N
ARG
X
126
−5.352
18.218
26.934
1.00
64.76
N

ATOM
908
CA
ARG
X
126
−4.440
17.315
27.626
1.00
64.76
C

ATOM
909
C
ARG
X
126
−4.852
17.059
29.069
1.00
64.76
C

ATOM
910
O
ARG
X
126
−4.330
16.144
29.719
1.00
64.76
O

ATOM
911
CB
ARG
X
126
−3.013
17.857
27.625
1.00
44.07
C

ATOM
912
CG
ARG
X
126
−2.272
17.627
26.344
1.00
44.07
C

ATOM
913
CD
ARG
X
126
−2.649
18.658
25.291
1.00
44.07
C

ATOM
914
NE
ARG
X
126
−1.742
19.806
25.268
1.00
44.07
N

ATOM
915
CZ
ARG
X
126
−0.417
19.716
25.322
1.00
44.07
C

ATOM
916
NH1
ARG
X
126
0.172
18.535
25.419
1.00
44.07
N

ATOM
917
NH2
ARG
X
126
0.324
20.805
25.243
1.00
44.07
N

ATOM
918
N
HIS
X
127
−5.768
17.873
29.586
1.00
61.54
N

ATOM
919
CA
HIS
X
127
−6.202
17.674
30.954
1.00
61.54
C

ATOM
920
C
HIS
X
127
−7.577
17.054
31.054
1.00
61.54
C

ATOM
921
O
HIS
X
127
−8.510
17.614
31.633
1.00
61.54
O

ATOM
922
CB
HIS
X
127
−6.154
18.967
31.759
1.00
99.00
C

ATOM
923
CG
HIS
X
127
−5.643
18.768
33.151
1.00
99.00
C

ATOM
924
ND1
HIS
X
127
−4.333
18.429
33.416
1.00
99.00
N

ATOM
925
CD2
HIS
X
127
−6.277
18.778
34.347
1.00
99.00
C

ATOM
926
CE1
HIS
X
127
−4.184
18.235
34.714
1.00
99.00
C

ATOM
927
NE2
HIS
X
127
−5.349
18.440
35.301
1.00
99.00
N

ATOM
928
N
CYS
X
128
−7.675
15.881
30.451
1.00
66.83
N

ATOM
929
CA
CYS
X
128
−8.870
15.068
30.466
1.00
66.83
C

ATOM
930
C
CYS
X
128
−8.256
13.704
30.661
1.00
66.83
C

ATOM
931
O
CYS
X
128
−8.936
12.728
30.954
1.00
66.83
O

ATOM
932
CB
CYS
X
128
−9.621
15.162
29.141
1.00
99.00
C

ATOM
933
SG
CYS
X
128
−10.751
16.576
29.066
1.00
99.00
S

ATOM
934
N
GLY
X
129
−6.936
13.675
30.503
1.00
72.04
N

ATOM
935
CA
GLY
X
129
−6.165
12.462
30.684
1.00
72.04
C

ATOM
936
C
GLY
X
129
−6.580
11.275
29.849
1.00
72.04
C

ATOM
937
O
GLY
X
129
−5.813
10.814
29.010
1.00
72.04
O

ATOM
938
N
ILE
X
130
−7.786
10.773
30.089
1.00
37.73
N

ATOM
939
CA
ILE
X
130
−8.290
9.630
29.360
1.00
37.73
C

ATOM
940
C
ILE
X
130
−9.574
9.940
28.604
1.00
37.73
C

ATOM
941
O
ILE
X
130
−10.493
10.543
29.148
1.00
37.73
O

ATOM
942
CB
ILE
X
130
−8.562
8.450
30.307
1.00
63.78
C

ATOM
943
CG1
ILE
X
130
−8.890
7.200
29.489
1.00
63.78
C

ATOM
944
CG2
ILE
X
130
−9.723
8.781
31.233
1.00
63.78
C

ATOM
945
CD1
ILE
X
130
−9.015
5.949
30.317
1.00
63.78
C

ATOM
946
N
MET
X
131
−9.639
9.501
27.351
1.00
69.22
N

ATOM
947
CA
MET
X
131
−10.819
9.721
26.519
1.00
69.22
C

ATOM
948
C
MET
X
131
−10.923
8.658
25.440
1.00
69.22
C

ATOM
949
O
MET
X
131
−9.938
8.001
25.102
1.00
69.22
O

ATOM
950
CB
MET
X
131
−10.747
11.083
25.839
1.00
74.80
C

ATOM
951
CG
MET
X
131
−9.551
11.220
24.922
1.00
74.80
C

ATOM
952
SD
MET
X
131
−9.720
12.587
23.787
1.00
74.80
S

ATOM
953
CE
MET
X
131
−8.990
13.896
24.741
1.00
74.80
C

ATOM
954
N
ASP
X
132
−12.125
8.501
24.899
1.00
63.49
N

ATOM
955
CA
ASP
X
132
−12.368
7.541
23.830
1.00
63.49
C

ATOM
956
C
ASP
X
132
−11.990
8.165
22.486
1.00
63.49
C

ATOM
957
O
ASP
X
132
−12.501
9.223
22.112
1.00
63.49
O

ATOM
958
CB
ASP
X
132
−13.849
7.129
23.783
1.00
56.30
C

ATOM
959
CG
ASP
X
132
−14.245
6.214
24.923
1.00
56.30
C

ATOM
960
OD1
ASP
X
132
−13.497
5.257
25.198
1.00
56.30
O

ATOM
961
OD2
ASP
X
132
−15.310
6.438
25.536
1.00
56.30
O

ATOM
962
N
VAL
X
133
−11.090
7.513
21.764
1.00
49.41
N

ATOM
963
CA
VAL
X
133
−10.678
7.994
20.454
1.00
49.41
C

ATOM
964
C
VAL
X
133
−11.121
7.015
19.361
1.00
49.41
C

ATOM
965
O
VAL
X
133
−11.863
6.071
19.626
1.00
49.41
O

ATOM
966
CB
VAL
X
133
−9.151
8.168
20.385
1.00
47.86
C

ATOM
967
CG1
VAL
X
133
−8.751
9.469
21.049
1.00
47.86
C

ATOM
968
CG2
VAL
X
133
−8.468
7.000
21.066
1.00
47.86
C

ATOM
969
N
GLU
X
134
−10.679
7.257
18.133
1.00
37.50
N

ATOM
970
CA
GLU
X
134
−11.005
6.382
17.013
1.00
37.50
C

ATOM
971
C
GLU
X
134
−9.755
6.207
16.186
1.00
37.50
C

ATOM
972
O
GLU
X
134
−8.998
7.159
16.016
1.00
37.50
O

ATOM
973
CB
GLU
X
134
−12.116
6.982
16.166
1.00
39.35
C

ATOM
974
CG
GLU
X
134
−13.483
6.443
16.519
1.00
39.35
C

ATOM
975
CD
GLU
X
134
−14.587
7.108
15.735
1.00
39.35
C

ATOM
976
OE1
GLU
X
134
−14.347
7.434
14.553
1.00
39.35
O

ATOM
977
OE2
GLU
X
134
−15.691
7.295
16.298
1.00
39.35
O

ATOM
978
N
PHE
X
135
−9.521
5.004
15.674
1.00
41.16
N

ATOM
979
CA
PHE
X
135
−8.309
4.788
14.893
1.00
41.16
C

ATOM
980
C
PHE
X
135
−8.401
3.698
13.832
1.00
41.16
C

ATOM
981
O
PHE
X
135
−9.391
2.972
13.732
1.00
41.16
O

ATOM
982
CB
PHE
X
135
−7.165
4.406
15.817
1.00
16.80
C

ATOM
983
CG
PHE
X
135
−7.208
2.972
16.236
1.00
16.80
C

ATOM
984
CD1
PHE
X
135
−8.232
2.509
17.050
1.00
16.80
C

ATOM
985
CD2
PHE
X
135
−6.276
2.064
15.737
1.00
16.80
C

ATOM
986
CE1
PHE
X
135
−8.332
1.163
17.359
1.00
16.80
C

ATOM
987
CE2
PHE
X
135
−6.365
0.715
16.037
1.00
16.80
C

ATOM
988
CZ
PHE
X
135
−7.393
0.258
16.848
1.00
16.80
C

ATOM
989
N
ARG
X
136
−7.328
3.591
13.059
1.00
11.01
N

ATOM
990
CA
ARG
X
136
−7.210
2.586
12.039
1.00
11.01
C

ATOM
991
C
ARG
X
136
−5.763
2.595
11.607
1.00
11.01
C

ATOM
992
O
ARG
X
136
−5.156
3.644
11.524
1.00
11.01
O

ATOM
993
CB
ARG
X
136
−8.135
2.889
10.858
1.00
49.76
C

ATOM
994
CG
ARG
X
136
−7.888
4.187
10.107
1.00
49.76
C

ATOM
995
CD
ARG
X
136
−8.927
4.292
9.003
1.00
49.76
C

ATOM
996
NE
ARG
X
136
−9.281
5.662
8.636
1.00
49.76
N

ATOM
997
CZ
ARG
X
136
−8.576
6.427
7.814
1.00
49.76
C

ATOM
998
NH1
ARG
X
136
−7.465
5.971
7.255
1.00
49.76
N

ATOM
999
NH2
ARG
X
136
−8.988
7.651
7.547
1.00
49.76
N

ATOM
1000
N
ARG
X
137
−5.178
1.435
11.366
1.00
24.96
N

ATOM
1001
CA
ARG
X
137
−3.788
1.417
10.944
1.00
24.96
C

ATOM
1002
C
ARG
X
137
−3.711
2.019
9.538
1.00
24.96
C

ATOM
1003
O
ARG
X
137
−4.736
2.177
8.872
1.00
24.96
O

ATOM
1004
CB
ARG
X
137
−3.256
−0.019
10.952
1.00
17.89
C

ATOM
1005
CG
ARG
X
137
−1.799
−0.133
10.557
1.00
17.89
C

ATOM
1006
CD
ARG
X
137
−1.233
−1.521
10.847
1.00
17.89
C

ATOM
1007
NE
ARG
X
137
0.169
−1.625
10.438
1.00
17.89
N

ATOM
1008
CZ
ARG
X
137
0.861
−2.757
10.472
1.00
17.89
C

ATOM
1009
NH1
ARG
X
137
0.278
−3.869
10.897
1.00
17.89
N

ATOM
1010
NH2
ARG
X
137
2.125
−2.779
10.073
1.00
17.89
N

ATOM
1011
N
VAL
X
138
−2.505
2.362
9.093
1.00
25.83
N

ATOM
1012
CA
VAL
X
138
−2.310
2.951
7.780
1.00
25.83
C

ATOM
1013
C
VAL
X
138
−0.899
2.646
7.314
1.00
25.83
C

ATOM
1014
O
VAL
X
138
−0.034
2.325
8.113
1.00
25.83
O

ATOM
1015
CB
VAL
X
138
−2.465
4.491
7.815
1.00
15.38
C

ATOM
1016
CG1
VAL
X
138
−3.714
4.875
8.569
1.00
15.38
C

ATOM
1017
CG2
VAL
X
138
−1.243
5.137
8.463
1.00
15.38
C

ATOM
1018
N
ARG
X
139
−0.650
2.725
6.022
1.00
43.70
N

ATOM
1019
CA
ARG
X
139
0.697
2.481
5.569
1.00
43.70
C

ATOM
1020
C
ARG
X
139
1.483
3.669
6.126
1.00
43.70
C

ATOM
1021
O
ARG
X
139
0.890
4.660
6.537
1.00
43.70
O

ATOM
1022
CB
ARG
X
139
0.733
2.436
4.043
1.00
50.01
C

ATOM
1023
CG
ARG
X
139
−0.096
1.301
3.464
1.00
50.01
C

ATOM
1024
CD
ARG
X
139
0.066
1.214
1.961
1.00
50.01
C

ATOM
1025
NE
ARG
X
139
0.028
−0.165
1.470
1.00
50.01
N

ATOM
1026
CZ
ARG
X
139
−1.074
−0.896
1.362
1.00
50.01
C

ATOM
1027
NH1
ARG
X
139
−2.246
−0.385
1.705
1.00
50.01
N

ATOM
1028
NH2
ARG
X
139
−0.998
−2.140
0.916
1.00
50.01
N

ATOM
1029
N
CYS
X
140
2.804
3.578
6.172
1.00
28.61
N

ATOM
1030
CA
CYS
X
140
3.580
4.684
6.702
1.00
28.61
C

ATOM
1031
C
CYS
X
140
4.266
5.422
5.587
1.00
28.61
C

ATOM
1032
O
CYS
X
140
4.840
4.807
4.698
1.00
28.61
O

ATOM
1033
CB
CYS
X
140
4.671
4.202
7.650
1.00
34.90
C

ATOM
1034
SG
CYS
X
140
4.213
3.206
9.085
1.00
34.90
S

ATOM
1035
N
LYS
X
141
4.259
6.744
5.672
1.00
40.28
N

ATOM
1036
CA
LYS
X
141
4.885
7.581
4.668
1.00
40.28
C

ATOM
1037
C
LYS
X
141
5.914
8.457
5.346
1.00
40.28
C

ATOM
1038
O
LYS
X
141
5.638
9.067
6.376
1.00
40.28
O

ATOM
1039
CB
LYS
X
141
3.803
8.423
3.986
1.00
85.00
C

ATOM
1040
CG
LYS
X
141
4.259
9.683
3.267
1.00
85.00
C

ATOM
1041
CD
LYS
X
141
3.512
10.896
3.845
1.00
85.00
C

ATOM
1042
CE
LYS
X
141
3.441
12.087
2.887
1.00
85.00
C

ATOM
1043
NZ
LYS
X
141
2.483
11.883
1.758
1.00
85.00
N

ATOM
1044
N
TYR
X
142
7.111
8.497
4.782
1.00
44.40
N

ATOM
1045
CA
TYR
X
142
8.200
9.320
5.310
1.00
44.40
C

ATOM
1046
C
TYR
X
142
8.589
10.333
4.231
1.00
44.40
C

ATOM
1047
O
TYR
X
142
8.116
10.258
3.098
1.00
44.40
O

ATOM
1048
CB
TYR
X
142
9.429
8.451
5.641
1.00
63.47
C

ATOM
1049
CG
TYR
X
142
9.331
7.661
6.927
1.00
63.47
C

ATOM
1050
CD1
TYR
X
142
9.514
8.276
8.161
1.00
63.47
C

ATOM
1051
CD2
TYR
X
142
9.015
6.308
6.912
1.00
63.47
C

ATOM
1052
CE1
TYR
X
142
9.378
7.564
9.348
1.00
63.47
C

ATOM
1053
CE2
TYR
X
142
8.876
5.588
8.090
1.00
63.47
C

ATOM
1054
CZ
TYR
X
142
9.056
6.221
9.307
1.00
63.47
C

ATOM
1055
OH
TYR
X
142
8.892
5.515
10.481
1.00
63.47
O

ATOM
1056
N
PRO
X
143
9.438
11.309
4.575
1.00
58.88
N

ATOM
1057
CA
PRO
X
143
9.821
12.267
3.542
1.00
58.88
C

ATOM
1058
C
PRO
X
143
10.342
11.480
2.342
1.00
58.88
C

ATOM
1059
O
PRO
X
143
10.807
10.350
2.483
1.00
58.88
O

ATOM
1060
CB
PRO
X
143
10.901
13.082
4.234
1.00
81.89
C

ATOM
1061
CG
PRO
X
143
10.376
13.156
5.632
1.00
81.89
C

ATOM
1062
CD
PRO
X
143
9.957
11.724
5.889
1.00
81.89
C

ATOM
1063
N
ALA
X
144
10.265
12.071
1.160
1.00
30.74
N

ATOM
1064
CA
ALA
X
144
10.704
11.382
−0.041
1.00
30.74
C

ATOM
1065
C
ALA
X
144
12.199
11.199
−0.068
1.00
30.74
C

ATOM
1066
O
ALA
X
144
12.951
12.033
0.421
1.00
30.74
O

ATOM
1067
CB
ALA
X
144
10.238
12.141
−1.283
1.00
49.37
C

ATOM
1068
N
GLY
X
145
12.630
10.093
−0.651
1.00
48.20
N

ATOM
1069
CA
GLY
X
145
14.047
9.822
−0.738
1.00
48.20
C

ATOM
1070
C
GLY
X
145
14.568
9.167
0.516
1.00
48.20
C

ATOM
1071
O
GLY
X
145
15.619
8.543
0.479
1.00
48.20
O

ATOM
1072
N
GLN
X
146
13.832
9.308
1.618
1.00
42.12
N

ATOM
1073
CA
GLN
X
146
14.215
8.739
2.913
1.00
42.12
C

ATOM
1074
C
GLN
X
146
14.158
7.220
2.904
1.00
42.12
C

ATOM
1075
O
GLN
X
146
13.075
6.641
2.993
1.00
42.12
O

ATOM
1076
CB
GLN
X
146
13.285
9.265
4.009
1.00
58.85
C

ATOM
1077
CG
GLN
X
146
13.442
8.563
5.347
1.00
58.85
C

ATOM
1078
CD
GLN
X
146
14.754
8.888
6.018
1.00
58.85
C

ATOM
1079
OE1
GLN
X
146
14.943
9.989
6.537
1.00
58.85
O

ATOM
1080
NE2
GLN
X
146
15.675
7.936
6.003
1.00
58.85
N

ATOM
1081
N
LYS
X
147
15.319
6.577
2.818
1.00
48.15
N

ATOM
1082
CA
LYS
X
147
15.373
5.119
2.784
1.00
48.15
C

ATOM
1083
C
LYS
X
147
15.513
4.518
4.184
1.00
48.15
C

ATOM
1084
O
LYS
X
147
15.515
5.243
5.178
1.00
48.15
O

ATOM
1085
CB
LYS
X
147
16.522
4.659
1.885
1.00
50.56
C

ATOM
1086
CG
LYS
X
147
16.720
5.537
0.650
1.00
50.56
C

ATOM
1087
CD
LYS
X
147
15.428
5.738
−0.166
1.00
50.56
C

ATOM
1088
CE
LYS
X
147
15.290
4.724
−1.284
1.00
50.56
C

ATOM
1089
NZ
LYS
X
147
16.461
4.788
−2.198
1.00
50.56
N

ATOM
1090
N
ILE
X
148
15.619
3.193
4.257
1.00
55.60
N

ATOM
1091
CA
ILE
X
148
15.732
2.504
5.539
1.00
55.60
C

ATOM
1092
C
ILE
X
148
17.118
2.575
6.160
1.00
55.60
C

ATOM
1093
O
ILE
X
148
18.124
2.319
5.497
1.00
55.60
O

ATOM
1094
CB
ILE
X
148
15.333
1.040
5.408
1.00
49.44
C

ATOM
1095
CG1
ILE
X
148
13.836
0.943
5.139
1.00
49.44
C

ATOM
1096
CG2
ILE
X
148
15.678
0.296
6.673
1.00
49.44
C

ATOM
1097
CD1
ILE
X
148
13.346
−0.468
4.945
1.00
49.44
C

ATOM
1098
N
VAL
X
149
17.154
2.902
7.450
1.00
57.64
N

ATOM
1099
CA
VAL
X
149
18.412
3.031
8.168
1.00
57.64
C

ATOM
1100
C
VAL
X
149
18.690
1.992
9.254
1.00
57.64
C

ATOM
1101
O
VAL
X
149
17.831
1.691
10.087
1.00
57.64
O

ATOM
1102
CB
VAL
X
149
18.515
4.414
8.797
1.00
56.80
C

ATOM
1103
CG1
VAL
X
149
19.850
4.565
9.494
1.00
56.80
C

ATOM
1104
CG2
VAL
X
149
18.351
5.466
7.722
1.00
56.80
C

ATOM
1105
N
PHE
X
150
19.910
1.459
9.223
1.00
43.64
N

ATOM
1106
CA
PHE
X
150
20.376
0.470
10.193
1.00
43.64
C

ATOM
1107
C
PHE
X
150
21.378
1.098
11.164
1.00
43.64
C

ATOM
1108
O
PHE
X
150
22.375
1.699
10.751
1.00
43.64
O

ATOM
1109
CB
PHE
X
150
21.056
−0.697
9.488
1.00
68.61
C

ATOM
1110
CG
PHE
X
150
20.250
−1.282
8.375
1.00
68.61
C

ATOM
1111
CD1
PHE
X
150
20.217
−0.666
7.125
1.00
68.61
C

ATOM
1112
CD2
PHE
X
150
19.525
−2.457
8.568
1.00
68.61
C

ATOM
1113
CE1
PHE
X
150
19.478
−1.212
6.084
1.00
68.61
C

ATOM
1114
CE2
PHE
X
150
18.781
−3.012
7.533
1.00
68.61
C

ATOM
1115
CZ
PHE
X
150
18.758
−2.387
6.288
1.00
68.61
C

ATOM
1116
N
HIS
X
151
21.109
0.944
12.455
1.00
51.27
N

ATOM
1117
CA
HIS
X
151
21.975
1.489
13.497
1.00
51.27
C

ATOM
1118
C
HIS
X
151
22.589
0.356
14.313
1.00
51.27
C

ATOM
1119
O
HIS
X
151
21.877
−0.536
14.777
1.00
51.27
O

ATOM
1120
CB
HIS
X
151
21.166
2.398
14.421
1.00
61.52
C

ATOM
1121
CG
HIS
X
151
21.981
3.059
15.486
1.00
61.52
C

ATOM
1122
ND1
HIS
X
151
22.887
2.375
16.266
1.00
61.52
N

ATOM
1123
CD2
HIS
X
151
22.000
4.339
15.923
1.00
61.52
C

ATOM
1124
CE1
HIS
X
151
23.429
3.205
17.138
1.00
61.52
C

ATOM
1125
NE2
HIS
X
151
22.908
4.403
16.951
1.00
61.52
N

ATOM
1126
N
ILE
X
152
23.909
0.399
14.487
1.00
66.30
N

ATOM
1127
CA
ILE
X
152
24.622
−0.620
15.256
1.00
66.30
C

ATOM
1128
C
ILE
X
152
24.543
−0.279
16.741
1.00
66.30
C

ATOM
1129
O
ILE
X
152
24.780
0.862
17.125
1.00
66.30
O

ATOM
1130
CB
ILE
X
152
26.098
−0.680
14.852
1.00
59.48
C

ATOM
1131
CG1
ILE
X
152
26.218
−0.727
13.328
1.00
59.48
C

ATOM
1132
CG2
ILE
X
152
26.743
−1.914
15.445
1.00
59.48
C

ATOM
1133
CD1
ILE
X
152
25.636
−1.978
12.698
1.00
59.48
C

ATOM
1134
N
GLU
X
153
24.217
−1.262
17.577
1.00
70.76
N

ATOM
1135
CA
GLU
X
153
24.103
−1.017
19.014
1.00
70.76
C

ATOM
1136
C
GLU
X
153
25.381
−1.317
19.815
1.00
70.76
C

ATOM
1137
O
GLU
X
153
26.319
−1.946
19.306
1.00
70.76
O

ATOM
1138
CB
GLU
X
153
22.929
−1.815
19.587
1.00
42.71
C

ATOM
1139
CG
GLU
X
153
21.567
−1.481
18.986
1.00
42.71
C

ATOM
1140
CD
GLU
X
153
21.257
0.010
18.998
1.00
42.71
C

ATOM
1141
OE1
GLU
X
153
21.934
0.764
18.260
1.00
42.71
O

ATOM
1142
OE2
GLU
X
153
20.343
0.435
19.742
1.00
42.71
O

ATOM
1143
N
LYS
X
154
25.406
−0.859
21.069
1.00
71.01
N

ATOM
1144
CA
LYS
X
154
26.555
−1.046
21.960
1.00
71.01
C

ATOM
1145
C
LYS
X
154
26.836
−2.519
22.275
1.00
71.01
C

ATOM
1146
O
LYS
X
154
26.030
−3.192
22.908
1.00
71.01
O

ATOM
1147
CB
LYS
X
154
26.345
−0.253
23.256
1.00
69.14
C

ATOM
1148
CG
LYS
X
154
25.140
−0.653
24.113
1.00
69.14
C

ATOM
1149
CD
LYS
X
154
23.783
−0.316
23.477
1.00
69.14
C

ATOM
1150
CE
LYS
X
154
22.667
−0.349
24.540
1.00
69.14
C

ATOM
1151
NZ
LYS
X
154
21.294
−0.587
24.014
1.00
69.14
N

ATOM
1152
N
GLY
X
155
28.000
−3.004
21.850
1.00
74.77
N

ATOM
1153
CA
GLY
X
155
28.342
−4.405
22.049
1.00
74.77
C

ATOM
1154
C
GLY
X
155
28.940
−4.886
23.359
1.00
74.77
C

ATOM
1155
O
GLY
X
155
28.896
−4.195
24.379
1.00
74.77
O

ATOM
1156
N
CYS
X
156
29.488
−6.103
23.311
1.00
60.33
N

ATOM
1157
CA
CYS
X
156
30.128
−6.753
24.459
1.00
60.33
C

ATOM
1158
C
CYS
X
156
31.083
−7.845
23.972
1.00
60.33
C

ATOM
1159
O
CYS
X
156
32.055
−7.558
23.268
1.00
60.33
O

ATOM
1160
CB
CYS
X
156
29.077
−7.375
25.382
1.00
97.61
C

ATOM
1161
SG
CYS
X
156
28.139
−8.745
24.652
1.00
97.61
S

ATOM
1162
N
ASN
X
157
30.802
−9.093
24.353
1.00
99.00
N

ATOM
1163
CA
ASN
X
157
31.626
−10.228
23.945
1.00
99.00
C

ATOM
1164
C
ASN
X
157
31.942
−10.077
22.462
1.00
99.00
C

ATOM
1165
O
ASN
X
157
31.062
−9.773
21.656
1.00
99.00
O

ATOM
1166
CB
ASN
X
157
30.903
−11.568
24.191
1.00
99.00
C

ATOM
1167
CG
ASN
X
157
30.843
−11.955
25.677
1.00
99.00
C

ATOM
1168
OD1
ASN
X
157
30.135
−11.327
26.469
1.00
99.00
O

ATOM
1169
ND2
ASN
X
157
31.588
−12.996
26.052
1.00
99.00
N

ATOM
1170
N
PRO
X
158
33.211
−10.287
22.088
1.00
99.00
N

ATOM
1171
CA
PRO
X
158
33.707
−10.181
20.715
1.00
99.00
C

ATOM
1172
C
PRO
X
158
32.691
−10.389
19.593
1.00
99.00
C

ATOM
1173
O
PRO
X
158
32.405
−11.518
19.186
1.00
99.00
O

ATOM
1174
CB
PRO
X
158
34.835
−11.200
20.693
1.00
99.00
C

ATOM
1175
CG
PRO
X
158
35.453
−10.973
22.039
1.00
99.00
C

ATOM
1176
CD
PRO
X
158
34.241
−10.883
22.960
1.00
99.00
C

ATOM
1177
N
ASN
X
159
32.151
−9.276
19.109
1.00
98.30
N

ATOM
1178
CA
ASN
X
159
31.191
−9.279
18.016
1.00
98.30
C

ATOM
1179
C
ASN
X
159
30.003
−10.213
18.169
1.00
98.30
C

ATOM
1180
O
ASN
X
159
30.070
−11.388
17.800
1.00
98.30
O

ATOM
1181
CB
ASN
X
159
31.900
−9.610
16.703
1.00
99.00
C

ATOM
1182
CG
ASN
X
159
33.025
−8.645
16.388
1.00
99.00
C

ATOM
1183
OD1
ASN
X
159
32.799
−7.453
16.177
1.00
99.00
O

ATOM
1184
ND2
ASN
X
159
34.250
−9.157
16.357
1.00
99.00
N

ATOM
1185
N
TYR
X
160
28.917
−9.685
18.722
1.00
99.00
N

ATOM
1186
CA
TYR
X
160
27.686
−10.449
18.864
1.00
99.00
C

ATOM
1187
C
TYR
X
160
26.638
−9.595
18.140
1.00
99.00
C

ATOM
1188
O
TYR
X
160
25.451
−9.930
18.082
1.00
99.00
O

ATOM
1189
CB
TYR
X
160
27.316
−10.650
20.343
1.00
90.79
C

ATOM
1190
CG
TYR
X
160
26.677
−9.457
21.018
1.00
90.79
C

ATOM
1191
CD1
TYR
X
160
25.604
−9.627
21.897
1.00
90.79
C

ATOM
1192
CD2
TYR
X
160
27.131
−8.159
20.775
1.00
90.79
C

ATOM
1193
CE1
TYR
X
160
24.995
−8.534
22.514
1.00
90.79
C

ATOM
1194
CE2
TYR
X
160
26.529
−7.059
21.386
1.00
90.79
C

ATOM
1195
CZ
TYR
X
160
25.461
−7.252
22.252
1.00
90.79
C

ATOM
1196
OH
TYR
X
160
24.855
−6.164
22.841
1.00
90.79
O

ATOM
1197
N
LEU
X
161
27.124
−8.487
17.579
1.00
99.00
N

ATOM
1198
CA
LEU
X
161
26.328
−7.509
16.835
1.00
99.00
C

ATOM
1199
C
LEU
X
161
25.051
−7.014
17.498
1.00
99.00
C

ATOM
1200
O
LEU
X
161
25.016
−6.793
18.708
1.00
99.00
O

ATOM
1201
CB
LEU
X
161
26.002
−8.026
15.427
1.00
54.38
C

ATOM
1202
CG
LEU
X
161
26.665
−7.134
14.367
1.00
54.38
C

ATOM
1203
CD1
LEU
X
161
26.392
−7.653
12.971
1.00
54.38
C

ATOM
1204
CD2
LEU
X
161
26.152
−5.708
14.525
1.00
54.38
C

ATOM
1205
N
ALA
X
162
24.022
−6.827
16.674
1.00
66.37
N

ATOM
1206
CA
ALA
X
162
22.701
−6.337
17.073
1.00
66.37
C

ATOM
1207
C
ALA
X
162
22.420
−5.061
16.288
1.00
66.37
C

ATOM
1208
O
ALA
X
162
23.061
−4.029
16.501
1.00
66.37
O

ATOM
1209
CB
ALA
X
162
22.630
−6.054
18.575
1.00
75.27
C

ATOM
1210
N
VAL
X
163
21.459
−5.140
15.375
1.00
52.28
N

ATOM
1211
CA
VAL
X
163
21.098
−4.003
14.541
1.00
52.28
C

ATOM
1212
C
VAL
X
163
19.701
−3.488
14.809
1.00
52.28
C

ATOM
1213
O
VAL
X
163
18.766
−4.258
15.009
1.00
52.28
O

ATOM
1214
CB
VAL
X
163
21.143
−4.362
13.047
1.00
52.25
C

ATOM
1215
CG1
VAL
X
163
20.834
−3.139
12.219
1.00
52.25
C

ATOM
1216
CG2
VAL
X
163
22.495
−4.924
12.681
1.00
52.25
C

ATOM
1217
N
LEU
X
164
19.569
−2.173
14.816
1.00
57.49
N

ATOM
1218
CA
LEU
X
164
18.276
−1.551
14.999
1.00
57.49
C

ATOM
1219
C
LEU
X
164
17.901
−1.108
13.593
1.00
57.49
C

ATOM
1220
O
LEU
X
164
18.770
−0.718
12.812
1.00
57.49
O

ATOM
1221
CB
LEU
X
164
18.383
−0.330
15.905
1.00
33.33
C

ATOM
1222
CG
LEU
X
164
17.168
0.603
15.832
1.00
33.33
C

ATOM
1223
CD1
LEU
X
164
16.044
0.058
16.695
1.00
33.33
C

ATOM
1224
CD2
LEU
X
164
17.554
1.996
16.284
1.00
33.33
C

ATOM
1225
N
VAL
X
165
16.617
−1.181
13.262
1.00
40.21
N

ATOM
1226
CA
VAL
X
165
16.138
−0.768
11.946
1.00
40.21
C

ATOM
1227
C
VAL
X
165
15.061
0.286
12.109
1.00
40.21
C

ATOM
1228
O
VAL
X
165
14.048
0.046
12.756
1.00
40.21
O

ATOM
1229
CB
VAL
X
165
15.559
−1.956
11.183
1.00
42.21
C

ATOM
1230
CG1
VAL
X
165
16.648
−2.623
10.380
1.00
42.21
C

ATOM
1231
CG2
VAL
X
165
14.963
−2.956
12.161
1.00
42.21
C

ATOM
1232
N
LYS
X
166
15.279
1.456
11.529
1.00
42.70
N

ATOM
1233
CA
LYS
X
166
14.301
2.527
11.652
1.00
42.70
C

ATOM
1234
C
LYS
X
166
13.844
3.022
10.303
1.00
42.70
C

ATOM
1235
O
LYS
X
166
14.486
2.748
9.295
1.00
42.70
O

ATOM
1236
CB
LYS
X
166
14.897
3.720
12.387
1.00
55.19
C

ATOM
1237
CG
LYS
X
166
15.686
3.387
13.613
1.00
55.19
C

ATOM
1238
CD
LYS
X
166
16.028
4.659
14.369
1.00
55.19
C

ATOM
1239
CE
LYS
X
166
16.810
5.634
13.518
1.00
55.19
C

ATOM
1240
NZ
LYS
X
166
16.940
6.917
14.245
1.00
55.19
N

ATOM
1241
N
TYR
X
167
12.747
3.776
10.302
1.00
47.87
N

ATOM
1242
CA
TYR
X
167
12.210
4.373
9.079
1.00
47.87
C

ATOM
1243
C
TYR
X
167
11.809
3.349
8.041
1.00
47.87
C

ATOM
1244
O
TYR
X
167
12.330
3.345
6.927
1.00
47.87
O

ATOM
1245
CB
TYR
X
167
13.241
5.316
8.455
1.00
63.86
C

ATOM
1246
CG
TYR
X
167
13.829
6.310
9.422
1.00
63.86
C

ATOM
1247
CD1
TYR
X
167
14.972
7.029
9.093
1.00
63.86
C

ATOM
1248
CD2
TYR
X
167
13.255
6.525
10.672
1.00
63.86
C

ATOM
1249
CE1
TYR
X
167
15.532
7.932
9.984
1.00
63.86
C

ATOM
1250
CE2
TYR
X
167
13.807
7.424
11.567
1.00
63.86
C

ATOM
1251
CZ
TYR
X
167
14.944
8.123
11.219
1.00
63.86
C

ATOM
1252
OH
TYR
X
167
15.495
9.014
12.108
1.00
63.86
O

ATOM
1253
N
VAL
X
168
10.891
2.472
8.415
1.00
42.72
N

ATOM
1254
CA
VAL
X
168
10.428
1.461
7.496
1.00
42.72
C

ATOM
1255
C
VAL
X
168
9.031
1.880
7.078
1.00
42.72
C

ATOM
1256
O
VAL
X
168
8.087
1.824
7.859
1.00
42.72
O

ATOM
1257
CB
VAL
X
168
10.393
0.085
8.152
1.00
30.21
C

ATOM
1258
CG1
VAL
X
168
10.142
−0.959
7.102
1.00
30.21
C

ATOM
1259
CG2
VAL
X
168
11.713
−0.194
8.842
1.00
30.21
C

ATOM
1260
N
ALA
X
169
8.918
2.325
5.835
1.00
38.54
N

ATOM
1261
CA
ALA
X
169
7.649
2.779
5.307
1.00
38.54
C

ATOM
1262
C
ALA
X
169
6.695
1.645
4.985
1.00
38.54
C

ATOM
1263
O
ALA
X
169
7.091
0.506
4.804
1.00
38.54
O

ATOM
1264
CB
ALA
X
169
7.885
3.624
4.066
1.00
21.47
C

ATOM
1265
N
ASP
X
170
5.422
1.992
4.933
1.00
28.44
N

ATOM
1266
CA
ASP
X
170
4.354
1.077
4.598
1.00
28.44
C

ATOM
1267
C
ASP
X
170
3.917
0.242
5.744
1.00
28.44
C

ATOM
1268
O
ASP
X
170
3.105
0.691
6.544
1.00
28.44
O

ATOM
1269
CB
ASP
X
170
4.737
0.182
3.415
1.00
24.17
C

ATOM
1270
CG
ASP
X
170
3.569
−0.615
2.897
1.00
24.17
C

ATOM
1271
OD1
ASP
X
170
2.423
−0.165
3.077
1.00
24.17
O

ATOM
1272
OD2
ASP
X
170
3.805
−1.685
2.303
1.00
24.17
O

ATOM
1273
N
ASP
X
171
4.419
−0.983
5.823
1.00
30.15
N

ATOM
1274
CA
ASP
X
171
4.022
−1.846
6.922
1.00
30.15
C

ATOM
1275
C
ASP
X
171
4.545
−1.247
8.201
1.00
30.15
C

ATOM
1276
O
ASP
X
171
3.897
−1.330
9.230
1.00
30.15
O

ATOM
1277
CB
ASP
X
171
4.564
−3.253
6.736
1.00
44.91
C

ATOM
1278
CG
ASP
X
171
3.683
−4.090
5.848
1.00
44.91
C

ATOM
1279
OD1
ASP
X
171
2.453
−4.007
6.011
1.00
44.91
O

ATOM
1280
OD2
ASP
X
171
4.214
−4.830
5.000
1.00
44.91
O

ATOM
1281
N
GLY
X
172
5.712
−0.620
8.109
1.00
50.12
N

ATOM
1282
CA
GLY
X
172
6.311
0.019
9.257
1.00
50.12
C

ATOM
1283
C
GLY
X
172
7.092
−0.913
10.160
1.00
50.12
C

ATOM
1284
O
GLY
X
172
8.214
−0.604
10.559
1.00
50.12
O

ATOM
1285
N
ASP
X
173
6.508
−2.065
10.468
1.00
39.19
N

ATOM
1286
CA
ASP
X
173
7.127
−3.025
11.361
1.00
39.19
C

ATOM
1287
C
ASP
X
173
7.811
−4.206
10.703
1.00
39.19
C

ATOM
1288
O
ASP
X
173
7.370
−4.707
9.669
1.00
39.19
O

ATOM
1289
CB
ASP
X
173
6.077
−3.543
12.326
1.00
28.64
C

ATOM
1290
CG
ASP
X
173
4.857
−4.059
11.612
1.00
28.64
C

ATOM
1291
OD1
ASP
X
173
4.972
−4.405
10.420
1.00
28.64
O

ATOM
1292
OD2
ASP
X
173
3.786
−4.137
12.243
1.00
28.64
O

ATOM
1293
N
ILE
X
174
8.888
−4.658
11.340
1.00
59.27
N

ATOM
1294
CA
ILE
X
174
9.666
−5.800
10.876
1.00
59.27
C

ATOM
1295
C
ILE
X
174
9.296
−7.052
11.669
1.00
59.27
C

ATOM
1296
O
ILE
X
174
8.774
−6.966
12.774
1.00
59.27
O

ATOM
1297
CB
ILE
X
174
11.148
−5.531
11.049
1.00
32.86
C

ATOM
1298
CG1
ILE
X
174
11.563
−4.360
10.158
1.00
32.86
C

ATOM
1299
CG2
ILE
X
174
11.945
−6.767
10.719
1.00
32.86
C

ATOM
1300
CD1
ILE
X
174
11.525
−4.652
8.676
1.00
32.86
C

ATOM
1301
N
VAL
X
175
9.554
−8.217
11.091
1.00
33.73
N

ATOM
1302
CA
VAL
X
175
9.238
−9.478
11.743
1.00
33.73
C

ATOM
1303
C
VAL
X
175
10.327
−10.516
11.496
1.00
33.73
C

ATOM
1304
O
VAL
X
175
10.178
−11.685
11.849
1.00
33.73
O

ATOM
1305
CB
VAL
X
175
7.875
−10.044
11.259
1.00
28.91
C

ATOM
1306
CG1
VAL
X
175
6.745
−9.134
11.707
1.00
28.91
C

ATOM
1307
CG2
VAL
X
175
7.865
−10.175
9.760
1.00
28.91
C

ATOM
1308
N
LEU
X
176
11.425
−10.083
10.894
1.00
32.04
N

ATOM
1309
CA
LEU
X
176
12.534
−10.974
10.631
1.00
32.04
C

ATOM
1310
C
LEU
X
176
13.648
−10.275
9.895
1.00
32.04
C

ATOM
1311
O
LEU
X
176
13.410
−9.588
8.909
1.00
32.04
O

ATOM
1312
CB
LEU
X
176
12.083
−12.182
9.805
1.00
58.82
C

ATOM
1313
CG
LEU
X
176
13.235
−13.042
9.270
1.00
58.82
C

ATOM
1314
CD1
LEU
X
176
14.053
−13.581
10.434
1.00
58.82
C

ATOM
1315
CD2
LEU
X
176
12.694
−14.179
8.427
1.00
58.82
C

ATOM
1316
N
MET
X
177
14.870
−10.452
10.376
1.00
50.60
N

ATOM
1317
CA
MET
X
177
16.027
−9.867
9.728
1.00
50.60
C

ATOM
1318
C
MET
X
177
16.967
−11.047
9.526
1.00
50.60
C

ATOM
1319
O
MET
X
177
16.817
−12.067
10.195
1.00
50.60
O

ATOM
1320
CB
MET
X
177
16.664
−8.812
10.630
1.00
45.74
C

ATOM
1321
CG
MET
X
177
17.599
−7.862
9.902
1.00
45.74
C

ATOM
1322
SD
MET
X
177
18.263
−6.531
10.940
1.00
45.74
S

ATOM
1323
CE
MET
X
177
16.776
−5.732
11.441
1.00
45.74
C

ATOM
1324
N
GLU
X
178
17.908
−10.936
8.594
1.00
55.01
N

ATOM
1325
CA
GLU
X
178
18.851
−12.023
8.347
1.00
55.01
C

ATOM
1326
C
GLU
X
178
20.025
−11.540
7.532
1.00
55.01
C

ATOM
1327
O
GLU
X
178
19.942
−10.498
6.887
1.00
55.01
O

ATOM
1328
CB
GLU
X
178
18.168
−13.181
7.638
1.00
62.11
C

ATOM
1329
CG
GLU
X
178
17.396
−12.775
6.435
1.00
62.11
C

ATOM
1330
CD
GLU
X
178
16.243
−13.704
6.199
1.00
62.11
C

ATOM
1331
OE1
GLU
X
178
15.512
−13.996
7.170
1.00
62.11
O

ATOM
1332
OE2
GLU
X
178
16.060
−14.137
5.048
1.00
62.11
O

ATOM
1333
N
ILE
X
179
21.118
−12.300
7.554
1.00
70.48
N

ATOM
1334
CA
ILE
X
179
22.323
−11.885
6.849
1.00
70.48
C

ATOM
1335
C
ILE
X
179
22.946
−12.938
5.944
1.00
70.48
C

ATOM
1336
O
ILE
X
179
22.351
−13.970
5.659
1.00
70.48
O

ATOM
1337
CB
ILE
X
179
23.398
−11.421
7.868
1.00
99.00
C

ATOM
1338
CG1
ILE
X
179
22.735
−10.609
8.985
1.00
99.00
C

ATOM
1339
CG2
ILE
X
179
24.438
−10.545
7.186
1.00
99.00
C

ATOM
1340
CD1
ILE
X
179
23.707
−10.056
10.002
1.00
99.00
C

ATOM
1341
N
GLN
X
180
24.158
−12.634
5.491
1.00
99.00
N

ATOM
1342
CA
GLN
X
180
24.966
−13.494
4.632
1.00
99.00
C

ATOM
1343
C
GLN
X
180
26.225
−12.702
4.291
1.00
99.00
C

ATOM
1344
O
GLN
X
180
26.155
−11.653
3.646
1.00
99.00
O

ATOM
1345
CB
GLN
X
180
24.216
−13.858
3.344
1.00
99.00
C

ATOM
1346
CG
GLN
X
180
23.878
−12.680
2.445
1.00
99.00
C

ATOM
1347
CD
GLN
X
180
23.917
−13.042
0.971
1.00
99.00
C

ATOM
1348
OE1
GLN
X
180
23.293
−14.009
0.542
1.00
99.00
O

ATOM
1349
NE2
GLN
X
180
24.655
−12.261
0.188
1.00
99.00
N

ATOM
1350
N
ASP
X
181
27.378
−13.181
4.741
1.00
72.36
N

ATOM
1351
CA
ASP
X
181
28.612
−12.466
4.445
1.00
72.36
C

ATOM
1352
C
ASP
X
181
29.149
−12.860
3.080
1.00
72.36
C

ATOM
1353
O
ASP
X
181
28.543
−13.672
2.379
1.00
72.36
O

ATOM
1354
CB
ASP
X
181
29.683
−12.699
5.531
1.00
62.59
C

ATOM
1355
CG
ASP
X
181
29.954
−14.175
5.804
1.00
62.59
C

ATOM
1356
OD1
ASP
X
181
29.128
−14.825
6.484
1.00
62.59
O

ATOM
1357
OD2
ASP
X
181
30.999
−14.684
5.341
1.00
62.59
O

ATOM
1358
N
LYS
X
182
30.281
−12.272
2.707
1.00
99.00
N

ATOM
1359
CA
LYS
X
182
30.904
−12.552
1.421
1.00
99.00
C

ATOM
1360
C
LYS
X
182
31.287
−14.024
1.320
1.00
99.00
C

ATOM
1361
O
LYS
X
182
32.335
−14.440
1.821
1.00
99.00
O

ATOM
1362
CB
LYS
X
182
32.147
−11.681
1.236
1.00
97.92
C

ATOM
1363
CG
LYS
X
182
31.853
−10.191
1.240
1.00
97.92
C

ATOM
1364
CD
LYS
X
182
33.110
−9.380
0.978
1.00
97.92
C

ATOM
1365
CE
LYS
X
182
33.681
−9.663
−0.402
1.00
97.92
C

ATOM
1366
NZ
LYS
X
182
34.881
−8.833
−0.681
1.00
97.92
N

ATOM
1367
N
LEU
X
183
30.427
−14.801
0.668
1.00
99.00
N

ATOM
1368
CA
LEU
X
183
30.629
−16.232
0.475
1.00
99.00
C

ATOM
1369
C
LEU
X
183
29.986
−17.064
1.570
1.00
99.00
C

ATOM
1370
O
LEU
X
183
30.214
−16.843
2.763
1.00
99.00
O

ATOM
1371
CB
LEU
X
183
32.119
−16.565
0.353
1.00
99.00
C

ATOM
1372
CG
LEU
X
183
32.709
−16.093
−0.980
1.00
99.00
C

ATOM
1373
CD1
LEU
X
183
34.174
−16.474
−1.067
1.00
99.00
C

ATOM
1374
CD2
LEU
X
183
31.908
−16.709
−2.134
1.00
99.00
C

ATOM
1375
N
SER
X
184
29.174
−18.021
1.122
1.00
99.00
N

ATOM
1376
CA
SER
X
184
28.427
−18.943
1.971
1.00
99.00
C

ATOM
1377
C
SER
X
184
27.017
−18.402
2.157
1.00
99.00
C

ATOM
1378
O
SER
X
184
26.432
−18.520
3.232
1.00
99.00
O

ATOM
1379
CB
SER
X
184
29.107
−19.117
3.333
1.00
91.50
C

ATOM
1380
OG
SER
X
184
30.426
−19.613
3.183
1.00
91.50
O

ATOM
1381
N
ALA
X
185
26.479
−17.813
1.091
1.00
66.67
N

ATOM
1382
CA
ALA
X
185
25.133
−17.245
1.116
1.00
66.67
C

ATOM
1383
C
ALA
X
185
24.068
−18.284
1.472
1.00
66.67
C

ATOM
1384
O
ALA
X
185
24.283
−19.483
1.314
1.00
66.67
O

ATOM
1385
CB
ALA
X
185
24.814
−16.617
−0.231
1.00
66.28
C

ATOM
1386
N
GLU
X
186
22.924
−17.813
1.960
1.00
87.47
N

ATOM
1387
CA
GLU
X
186
21.808
−18.676
2.346
1.00
87.47
C

ATOM
1388
C
GLU
X
186
20.919
−17.957
3.357
1.00
87.47
C

ATOM
1389
O
GLU
X
186
19.931
−18.509
3.850
1.00
87.47
O

ATOM
1390
CB
GLU
X
186
22.311
−19.998
2.936
1.00
83.62
C

ATOM
1391
CG
GLU
X
186
23.364
−19.848
4.005
1.00
83.62
C

ATOM
1392
CD
GLU
X
186
24.134
−21.126
4.232
1.00
83.62
C

ATOM
1393
OE1
GLU
X
186
23.519
−22.115
4.684
1.00
83.62
O

ATOM
1394
OE2
GLU
X
186
25.354
−21.145
3.950
1.00
83.62
O

ATOM
1395
N
TRP
X
187
21.283
−16.716
3.659
1.00
71.32
N

ATOM
1396
CA
TRP
X
187
20.527
−15.890
4.589
1.00
71.32
C

ATOM
1397
C
TRP
X
187
20.250
−16.555
5.926
1.00
71.32
C

ATOM
1398
O
TRP
X
187
19.266
−17.283
6.071
1.00
71.32
O

ATOM
1399
CB
TRP
X
187
19.188
−15.473
3.972
1.00
72.32
C

ATOM
1400
CG
TRP
X
187
19.311
−14.731
2.683
1.00
72.32
C

ATOM
1401
CD1
TRP
X
187
18.936
−15.169
1.450
1.00
72.32
C

ATOM
1402
CD2
TRP
X
187
19.858
−13.421
2.495
1.00
72.32
C

ATOM
1403
NE1
TRP
X
187
19.215
−14.215
0.499
1.00
72.32
N

ATOM
1404
CE2
TRP
X
187
19.782
−13.130
1.114
1.00
72.32
C

ATOM
1405
CE3
TRP
X
187
20.405
−12.462
3.359
1.00
72.32
C

ATOM
1406
CZ2
TRP
X
187
20.235
−11.923
0.574
1.00
72.32
C

ATOM
1407
CZ3
TRP
X
187
20.855
−11.260
2.822
1.00
72.32
C

ATOM
1408
CH2
TRP
X
187
20.766
−11.002
1.441
1.00
72.32
C

ATOM
1409
N
LYS
X
188
21.112
−16.306
6.904
1.00
84.38
N

ATOM
1410
CA
LYS
X
188
20.903
−16.859
8.233
1.00
84.38
C

ATOM
1411
C
LYS
X
188
20.061
−15.849
8.989
1.00
84.38
C

ATOM
1412
O
LYS
X
188
20.342
−14.652
8.957
1.00
84.38
O

ATOM
1413
CB
LYS
X
188
22.228
−17.067
8.968
1.00
99.00
C

ATOM
1414
CG
LYS
X
188
22.052
−17.280
10.474
1.00
99.00
C

ATOM
1415
CD
LYS
X
188
23.371
−17.562
11.183
1.00
99.00
C

ATOM
1416
CE
LYS
X
188
23.195
−17.576
12.701
1.00
99.00
C

ATOM
1417
NZ
LYS
X
188
22.144
−18.530
13.146
1.00
99.00
N

ATOM
1418
N
PRO
X
189
19.012
−16.319
9.674
1.00
61.36
N

ATOM
1419
CA
PRO
X
189
18.094
−15.491
10.457
1.00
61.36
C

ATOM
1420
C
PRO
X
189
18.811
−14.582
11.449
1.00
61.36
C

ATOM
1421
O
PRO
X
189
20.022
−14.374
11.360
1.00
61.36
O

ATOM
1422
CB
PRO
X
189
17.220
−16.522
11.163
1.00
38.13
C

ATOM
1423
CG
PRO
X
189
17.155
−17.615
10.197
1.00
38.13
C

ATOM
1424
CD
PRO
X
189
18.595
−17.726
9.720
1.00
38.13
C

ATOM
1425
N
MET
X
190
18.048
−14.044
12.395
1.00
43.42
N

ATOM
1426
CA
MET
X
190
18.596
−13.163
13.412
1.00
43.42
C

ATOM
1427
C
MET
X
190
17.694
−13.209
14.632
1.00
43.42
C

ATOM
1428
O
MET
X
190
16.489
−13.442
14.515
1.00
43.42
O

ATOM
1429
CB
MET
X
190
18.669
−11.735
12.888
1.00
58.50
C

ATOM
1430
CG
MET
X
190
19.474
−11.581
11.623
1.00
58.50
C

ATOM
1431
SD
MET
X
190
21.062
−10.854
11.938
1.00
58.50
S

ATOM
1432
CE
MET
X
190
20.559
−9.277
12.584
1.00
58.50
C

ATOM
1433
N
LYS
X
191
18.281
−12.980
15.801
1.00
57.01
N

ATOM
1434
CA
LYS
X
191
17.521
−13.010
17.038
1.00
57.01
C

ATOM
1435
C
LYS
X
191
16.869
−11.679
17.282
1.00
57.01
C

ATOM
1436
O
LYS
X
191
17.504
−10.641
17.127
1.00
57.01
O

ATOM
1437
CB
LYS
X
191
18.420
−13.309
18.234
1.00
99.00
C

ATOM
1438
CG
LYS
X
191
19.173
−14.618
18.195
1.00
99.00
C

ATOM
1439
CD
LYS
X
191
19.786
−14.914
19.561
1.00
99.00
C

ATOM
1440
CE
LYS
X
191
20.654
−13.758
20.086
1.00
99.00
C

ATOM
1441
NZ
LYS
X
191
19.887
−12.549
20.511
1.00
99.00
N

ATOM
1442
N
LEU
X
192
15.602
−11.711
17.670
1.00
54.42
N

ATOM
1443
CA
LEU
X
192
14.886
−10.487
17.975
1.00
54.42
C

ATOM
1444
C
LEU
X
192
15.202
−10.166
19.416
1.00
54.42
C

ATOM
1445
O
LEU
X
192
14.400
−10.467
20.296
1.00
54.42
O

ATOM
1446
CB
LEU
X
192
13.376
−10.675
17.838
1.00
82.95
C

ATOM
1447
CG
LEU
X
192
12.559
−9.581
18.541
1.00
82.95
C

ATOM
1448
CD1
LEU
X
192
12.809
−8.234
17.888
1.00
82.95
C

ATOM
1449
CD2
LEU
X
192
11.092
−9.929
18.500
1.00
82.95
C

ATOM
1450
N
SER
X
193
16.368
−9.575
19.669
1.00
53.14
N

ATOM
1451
CA
SER
X
193
16.741
−9.235
21.039
1.00
53.14
C

ATOM
1452
C
SER
X
193
15.525
−8.577
21.688
1.00
53.14
C

ATOM
1453
O
SER
X
193
14.972
−9.097
22.651
1.00
53.14
O

ATOM
1454
CB
SER
X
193
17.931
−8.281
21.056
1.00
67.21
C

ATOM
1455
OG
SER
X
193
17.492
−6.939
21.063
1.00
67.21
O

ATOM
1456
N
TRP
X
194
15.090
−7.448
21.145
1.00
38.60
N

ATOM
1457
CA
TRP
X
194
13.918
−6.771
21.677
1.00
38.60
C

ATOM
1458
C
TRP
X
194
13.545
−5.569
20.806
1.00
38.60
C

ATOM
1459
O
TRP
X
194
14.420
−4.887
20.259
1.00
38.60
O

ATOM
1460
CB
TRP
X
194
14.169
−6.351
23.134
1.00
50.51
C

ATOM
1461
CG
TRP
X
194
14.507
−4.918
23.312
1.00
50.51
C

ATOM
1462
CD1
TRP
X
194
13.629
−3.872
23.411
1.00
50.51
C

ATOM
1463
CD2
TRP
X
194
15.817
−4.348
23.353
1.00
50.51
C

ATOM
1464
NE1
TRP
X
194
14.316
−2.682
23.508
1.00
50.51
N

ATOM
1465
CE2
TRP
X
194
15.659
−2.943
23.475
1.00
50.51
C

ATOM
1466
CE3
TRP
X
194
17.109
−4.884
23.298
1.00
50.51
C

ATOM
1467
CZ2
TRP
X
194
16.742
−2.070
23.539
1.00
50.51
C

ATOM
1468
CZ3
TRP
X
194
18.190
−4.015
23.361
1.00
50.51
C

ATOM
1469
CH2
TRP
X
194
17.997
−2.620
23.481
1.00
50.51
C

ATOM
1470
N
GLY
X
195
12.239
−5.331
20.679
1.00
45.90
N

ATOM
1471
CA
GLY
X
195
11.733
−4.228
19.878
1.00
45.90
C

ATOM
1472
C
GLY
X
195
12.161
−4.328
18.426
1.00
45.90
C

ATOM
1473
O
GLY
X
195
11.788
−5.260
17.715
1.00
45.90
O

ATOM
1474
N
ALA
X
196
12.953
−3.359
17.985
1.00
29.15
N

ATOM
1475
CA
ALA
X
196
13.446
−3.323
16.622
1.00
29.15
C

ATOM
1476
C
ALA
X
196
14.925
−3.663
16.628
1.00
29.15
C

ATOM
1477
O
ALA
X
196
15.644
−3.376
15.668
1.00
29.15
O

ATOM
1478
CB
ALA
X
196
13.225
−1.934
16.024
1.00
5.86
C

ATOM
1479
N
ILE
X
197
15.377
−4.261
17.730
1.00
53.88
N

ATOM
1480
CA
ILE
X
197
16.775
−4.655
17.880
1.00
53.88
C

ATOM
1481
C
ILE
X
197
16.911
−6.124
17.548
1.00
53.88
C

ATOM
1482
O
ILE
X
197
16.249
−6.963
18.149
1.00
53.88
O

ATOM
1483
CB
ILE
X
197
17.277
−4.468
19.312
1.00
40.28
C

ATOM
1484
CG1
ILE
X
197
16.994
−3.049
19.798
1.00
40.28
C

ATOM
1485
CG2
ILE
X
197
18.769
−4.746
19.358
1.00
40.28
C

ATOM
1486
CD1
ILE
X
197
17.877
−2.015
19.180
1.00
40.28
C

ATOM
1487
N
TRP
X
198
17.776
−6.430
16.593
1.00
36.85
N

ATOM
1488
CA
TRP
X
198
17.987
−7.807
16.170
1.00
36.85
C

ATOM
1489
C
TRP
X
198
19.419
−8.229
16.458
1.00
36.85
C

ATOM
1490
O
TRP
X
198
20.360
−7.479
16.213
1.00
36.85
O

ATOM
1491
CB
TRP
X
198
17.623
−7.953
14.680
1.00
40.41
C

ATOM
1492
CG
TRP
X
198
16.134
−7.773
14.466
1.00
40.41
C

ATOM
1493
CD1
TRP
X
198
15.428
−6.616
14.603
1.00
40.41
C

ATOM
1494
CD2
TRP
X
198
15.160
−8.808
14.248
1.00
40.41
C

ATOM
1495
NE1
TRP
X
198
14.082
−6.862
14.501
1.00
40.41
N

ATOM
1496
CE2
TRP
X
198
13.888
−8.199
14.284
1.00
40.41
C

ATOM
1497
CE3
TRP
X
198
15.241
−10.194
14.038
1.00
40.41
C

ATOM
1498
CZ2
TRP
X
198
12.700
−8.927
14.118
1.00
40.41
C

ATOM
1499
CZ3
TRP
X
198
14.062
−10.918
13.876
1.00
40.41
C

ATOM
1500
CH2
TRP
X
198
12.809
−10.282
13.919
1.00
40.41
C

ATOM
1501
N
ARG
X
199
19.578
−9.438
16.982
1.00
72.25
N

ATOM
1502
CA
ARG
X
199
20.891
−9.932
17.369
1.00
72.25
C

ATOM
1503
C
ARG
X
199
21.432
−11.084
16.542
1.00
72.25
C

ATOM
1504
O
ARG
X
199
20.686
−11.937
16.062
1.00
72.25
O

ATOM
1505
CB
ARG
X
199
20.848
−10.362
18.835
1.00
99.00
C

ATOM
1506
CG
ARG
X
199
22.185
−10.374
19.537
1.00
99.00
C

ATOM
1507
CD
ARG
X
199
22.349
−9.125
20.397
1.00
99.00
C

ATOM
1508
NE
ARG
X
199
21.267
−8.987
21.375
1.00
99.00
N

ATOM
1509
CZ
ARG
X
199
21.233
−8.071
22.342
1.00
99.00
C

ATOM
1510
NH1
ARG
X
199
22.223
−7.200
22.476
1.00
99.00
N

ATOM
1511
NH2
ARG
X
199
20.202
−8.023
23.177
1.00
99.00
N

ATOM
1512
N
MET
X
200
22.752
−11.097
16.407
1.00
88.97
N

ATOM
1513
CA
MET
X
200
23.469
−12.129
15.677
1.00
88.97
C

ATOM
1514
C
MET
X
200
24.538
−12.663
16.626
1.00
88.97
C

ATOM
1515
O
MET
X
200
25.643
−12.128
16.677
1.00
88.97
O

ATOM
1516
CB
MET
X
200
24.124
−11.527
14.434
1.00
92.06
C

ATOM
1517
CG
MET
X
200
25.079
−12.460
13.714
1.00
92.06
C

ATOM
1518
SD
MET
X
200
24.329
−14.048
13.334
1.00
92.06
S

ATOM
1519
CE
MET
X
200
23.717
−13.762
11.661
1.00
92.06
C

ATOM
1520
N
ASP
X
201
24.200
−13.709
17.379
1.00
69.71
N

ATOM
1521
CA
ASP
X
201
25.112
−14.321
18.357
1.00
69.71
C

ATOM
1522
C
ASP
X
201
26.523
−14.561
17.832
1.00
69.71
C

ATOM
1523
O
ASP
X
201
27.489
−13.970
18.314
1.00
69.71
O

ATOM
1524
CB
ASP
X
201
24.538
−15.650
18.836
1.00
99.00
C

ATOM
1525
CG
ASP
X
201
23.074
−15.553
19.171
1.00
99.00
C

ATOM
1526
OD1
ASP
X
201
22.300
−15.117
18.293
1.00
99.00
O

ATOM
1527
OD2
ASP
X
201
22.697
−15.913
20.304
1.00
99.00
O

ATOM
1528
N
THR
X
202
26.627
−15.450
16.853
1.00
93.73
N

ATOM
1529
CA
THR
X
202
27.903
−15.787
16.242
1.00
93.73
C

ATOM
1530
C
THR
X
202
28.319
−14.692
15.260
1.00
93.73
C

ATOM
1531
O
THR
X
202
27.472
−14.086
14.604
1.00
93.73
O

ATOM
1532
CB
THR
X
202
27.800
−17.137
15.502
1.00
87.54
C

ATOM
1533
OG1
THR
X
202
29.031
−17.406
14.826
1.00
87.54
O

ATOM
1534
CG2
THR
X
202
26.656
−17.112
14.492
1.00
87.54
C

ATOM
1535
N
ALA
X
203
29.618
−14.432
15.159
1.00
96.00
N

ATOM
1536
CA
ALA
X
203
30.106
−13.399
14.251
1.00
96.00
C

ATOM
1537
C
ALA
X
203
31.551
−13.627
13.831
1.00
96.00
C

ATOM
1538
O
ALA
X
203
32.146
−12.782
13.160
1.00
96.00
O

ATOM
1539
CB
ALA
X
203
29.968
−12.030
14.900
1.00
53.35
C

ATOM
1540
N
LYS
X
204
32.112
−14.767
14.226
1.00
99.00
N

ATOM
1541
CA
LYS
X
204
33.489
−15.097
13.881
1.00
99.00
C

ATOM
1542
C
LYS
X
204
33.627
−15.336
12.377
1.00
99.00
C

ATOM
1543
O
LYS
X
204
34.480
−14.732
11.722
1.00
99.00
O

ATOM
1544
CB
LYS
X
204
33.950
−16.343
14.645
1.00
98.90
C

ATOM
1545
CG
LYS
X
204
33.280
−17.647
14.212
1.00
98.90
C

ATOM
1546
CD
LYS
X
204
31.800
−17.666
14.564
1.00
98.90
C

ATOM
1547
CE
LYS
X
204
31.103
−18.923
14.045
1.00
98.90
C

ATOM
1548
NZ
LYS
X
204
30.802
−18.870
12.586
1.00
98.90
N

ATOM
1549
N
ALA
X
205
32.782
−16.216
11.839
1.00
99.00
N

ATOM
1550
CA
ALA
X
205
32.798
−16.549
10.414
1.00
99.00
C

ATOM
1551
C
ALA
X
205
32.252
−15.391
9.589
1.00
99.00
C

ATOM
1552
O
ALA
X
205
32.922
−14.887
8.684
1.00
99.00
O

ATOM
1553
CB
ALA
X
205
31.971
−17.803
10.161
1.00
61.42
C

ATOM
1554
N
LEU
X
206
31.026
−14.983
9.903
1.00
71.39
N

ATOM
1555
CA
LEU
X
206
30.388
−13.873
9.217
1.00
71.39
C

ATOM
1556
C
LEU
X
206
31.364
−12.711
9.110
1.00
71.39
C

ATOM
1557
O
LEU
X
206
31.505
−11.940
10.057
1.00
71.39
O

ATOM
1558
CB
LEU
X
206
29.170
−13.401
10.005
1.00
80.72
C

ATOM
1559
CG
LEU
X
206
28.090
−14.427
10.319
1.00
80.72
C

ATOM
1560
CD1
LEU
X
206
27.042
−13.808
11.241
1.00
80.72
C

ATOM
1561
CD2
LEU
X
206
27.465
−14.907
9.019
1.00
80.72
C

ATOM
1562
N
LYS
X
207
32.041
−12.586
7.974
1.00
81.49
N

ATOM
1563
CA
LYS
X
207
32.979
−11.486
7.773
1.00
81.49
C

ATOM
1564
C
LYS
X
207
32.843
−10.942
6.356
1.00
81.49
C

ATOM
1565
O
LYS
X
207
32.485
−11.677
5.436
1.00
81.49
O

ATOM
1566
CB
LYS
X
207
34.419
−11.944
8.020
1.00
89.15
C

ATOM
1567
CG
LYS
X
207
34.689
−12.555
9.404
1.00
89.15
C

ATOM
1568
CD
LYS
X
207
34.048
−11.773
10.554
1.00
89.15
C

ATOM
1569
CE
LYS
X
207
34.328
−10.281
10.497
1.00
89.15
C

ATOM
1570
NZ
LYS
X
207
33.463
−9.553
11.466
1.00
89.15
N

ATOM
1571
N
GLY
X
208
33.130
−9.654
6.183
1.00
99.00
N

ATOM
1572
CA
GLY
X
208
33.010
−9.038
4.870
1.00
99.00
C

ATOM
1573
C
GLY
X
208
31.688
−8.300
4.756
1.00
99.00
C

ATOM
1574
O
GLY
X
208
30.742
−8.656
5.454
1.00
99.00
O

ATOM
1575
N
PRO
X
209
31.578
−7.275
3.892
1.00
80.63
N

ATOM
1576
CA
PRO
X
209
30.329
−6.517
3.739
1.00
80.63
C

ATOM
1577
C
PRO
X
209
29.106
−7.418
3.762
1.00
80.63
C

ATOM
1578
O
PRO
X
209
28.922
−8.253
2.878
1.00
80.63
O

ATOM
1579
CB
PRO
X
209
30.521
−5.816
2.404
1.00
87.69
C

ATOM
1580
CG
PRO
X
209
31.985
−5.507
2.429
1.00
87.69
C

ATOM
1581
CD
PRO
X
209
32.585
−6.814
2.920
1.00
87.69
C

ATOM
1582
N
PHE
X
210
28.271
−7.237
4.780
1.00
77.58
N

ATOM
1583
CA
PHE
X
210
27.084
−8.064
4.943
1.00
77.58
C

ATOM
1584
C
PHE
X
210
25.840
−7.498
4.301
1.00
77.58
C

ATOM
1585
O
PHE
X
210
25.646
−6.287
4.246
1.00
77.58
O

ATOM
1586
CB
PHE
X
210
26.787
−8.296
6.423
1.00
99.00
C

ATOM
1587
CG
PHE
X
210
27.988
−8.662
7.234
1.00
99.00
C

ATOM
1588
CD1
PHE
X
210
28.916
−7.693
7.601
1.00
99.00
C

ATOM
1589
CD2
PHE
X
210
28.192
−9.973
7.639
1.00
99.00
C

ATOM
1590
CE1
PHE
X
210
30.028
−8.021
8.361
1.00
99.00
C

ATOM
1591
CE2
PHE
X
210
29.300
−10.314
8.400
1.00
99.00
C

ATOM
1592
CZ
PHE
X
210
30.222
−9.335
8.763
1.00
99.00
C

ATOM
1593
N
SER
X
211
24.991
−8.409
3.839
1.00
83.22
N

ATOM
1594
CA
SER
X
211
23.728
−8.066
3.210
1.00
83.22
C

ATOM
1595
C
SER
X
211
22.641
−8.368
4.234
1.00
83.22
C

ATOM
1596
O
SER
X
211
22.730
−9.341
4.978
1.00
83.22
O

ATOM
1597
CB
SER
X
211
23.533
−8.914
1.960
1.00
78.36
C

ATOM
1598
OG
SER
X
211
24.712
−8.906
1.175
1.00
78.36
O

ATOM
1599
N
ILE
X
212
21.615
−7.534
4.277
1.00
61.70
N

ATOM
1600
CA
ILE
X
212
20.547
−7.727
5.237
1.00
61.70
C

ATOM
1601
C
ILE
X
212
19.205
−7.750
4.548
1.00
61.70
C

ATOM
1602
O
ILE
X
212
18.935
−6.913
3.691
1.00
61.70
O

ATOM
1603
CB
ILE
X
212
20.531
−6.598
6.257
1.00
67.55
C

ATOM
1604
CG1
ILE
X
212
21.906
−6.467
6.901
1.00
67.55
C

ATOM
1605
CG2
ILE
X
212
19.471
−6.863
7.300
1.00
67.55
C

ATOM
1606
CD1
ILE
X
212
22.007
−5.306
7.859
1.00
67.55
C

ATOM
1607
N
ARG
X
213
18.365
−8.702
4.937
1.00
56.98
N

ATOM
1608
CA
ARG
X
213
17.044
−8.832
4.356
1.00
56.98
C

ATOM
1609
C
ARG
X
213
15.976
−8.671
5.425
1.00
56.98
C

ATOM
1610
O
ARG
X
213
15.969
−9.402
6.410
1.00
56.98
O

ATOM
1611
CB
ARG
X
213
16.905
−10.185
3.684
1.00
56.46
C

ATOM
1612
CG
ARG
X
213
15.598
−10.344
2.977
1.00
56.46
C

ATOM
1613
CD
ARG
X
213
15.631
−11.537
2.065
1.00
56.46
C

ATOM
1614
NE
ARG
X
213
15.543
−12.800
2.784
1.00
56.46
N

ATOM
1615
CZ
ARG
X
213
15.651
−13.982
2.190
1.00
56.46
C

ATOM
1616
NH1
ARG
X
213
15.855
−14.040
0.880
1.00
56.46
N

ATOM
1617
NH2
ARG
X
213
15.544
−15.101
2.891
1.00
56.46
N

ATOM
1618
N
LEU
X
214
15.073
−7.712
5.215
1.00
44.58
N

ATOM
1619
CA
LEU
X
214
14.000
−7.415
6.155
1.00
44.58
C

ATOM
1620
C
LEU
X
214
12.691
−7.946
5.622
1.00
44.58
C

ATOM
1621
O
LEU
X
214
12.476
−7.962
4.418
1.00
44.58
O

ATOM
1622
CB
LEU
X
214
13.883
−5.907
6.329
1.00
52.06
C

ATOM
1623
CG
LEU
X
214
15.193
−5.169
6.600
1.00
52.06
C

ATOM
1624
CD1
LEU
X
214
14.992
−3.673
6.417
1.00
52.06
C

ATOM
1625
CD2
LEU
X
214
15.684
−5.500
8.007
1.00
52.06
C

ATOM
1626
N
THR
X
215
11.808
−8.371
6.515
1.00
35.43
N

ATOM
1627
CA
THR
X
215
10.515
−8.893
6.090
1.00
35.43
C

ATOM
1628
C
THR
X
215
9.438
−8.180
6.868
1.00
35.43
C

ATOM
1629
O
THR
X
215
9.335
−8.359
8.080
1.00
35.43
O

ATOM
1630
CB
THR
X
215
10.361
−10.394
6.391
1.00
36.84
C

ATOM
1631
OG1
THR
X
215
11.300
−11.151
5.625
1.00
36.84
O

ATOM
1632
CG2
THR
X
215
8.973
−10.845
6.044
1.00
36.84
C

ATOM
1633
N
SER
X
216
8.637
−7.379
6.176
1.00
18.36
N

ATOM
1634
CA
SER
X
216
7.564
−6.640
6.815
1.00
18.36
C

ATOM
1635
C
SER
X
216
6.573
−7.610
7.416
1.00
18.36
C

ATOM
1636
O
SER
X
216
6.673
−8.815
7.225
1.00
18.36
O

ATOM
1637
CB
SER
X
216
6.861
−5.763
5.797
1.00
48.23
C

ATOM
1638
OG
SER
X
216
6.404
−6.540
4.709
1.00
48.23
O

ATOM
1639
N
GLU
X
217
5.607
−7.079
8.147
1.00
33.43
N

ATOM
1640
CA
GLU
X
217
4.603
−7.909
8.782
1.00
33.43
C

ATOM
1641
C
GLU
X
217
3.864
−8.652
7.692
1.00
33.43
C

ATOM
1642
O
GLU
X
217
3.401
−9.755
7.910
1.00
33.43
O

ATOM
1643
CB
GLU
X
217
3.628
−7.034
9.591
1.00
30.80
C

ATOM
1644
CG
GLU
X
217
2.786
−7.756
10.654
1.00
30.80
C

ATOM
1645
CD
GLU
X
217
1.842
−6.805
11.392
1.00
30.80
C

ATOM
1646
OE1
GLU
X
217
2.216
−5.625
11.565
1.00
30.80
O

ATOM
1647
OE2
GLU
X
217
0.738
−7.226
11.812
1.00
30.80
O

ATOM
1648
N
SER
X
218
3.781
−8.041
6.514
1.00
50.46
N

ATOM
1649
CA
SER
X
218
3.075
−8.604
5.358
1.00
50.46
C

ATOM
1650
C
SER
X
218
3.734
−9.745
4.641
1.00
50.46
C

ATOM
1651
O
SER
X
218
3.058
−10.570
4.044
1.00
50.46
O

ATOM
1652
CB
SER
X
218
2.808
−7.523
4.326
1.00
27.90
C

ATOM
1653
OG
SER
X
218
1.700
−6.764
4.734
1.00
27.90
O

ATOM
1654
N
GLY
X
219
5.053
−9.778
4.674
1.00
34.08
N

ATOM
1655
CA
GLY
X
219
5.771
−10.827
3.992
1.00
34.08
C

ATOM
1656
C
GLY
X
219
6.695
−10.167
3.000
1.00
34.08
C

ATOM
1657
O
GLY
X
219
7.691
−10.750
2.559
1.00
34.08
O

ATOM
1658
N
LYS
X
220
6.373
−8.932
2.642
1.00
42.43
N

ATOM
1659
CA
LYS
X
220
7.207
−8.222
1.697
1.00
42.43
C

ATOM
1660
C
LYS
X
220
8.645
−8.261
2.221
1.00
42.43
C

ATOM
1661
O
LYS
X
220
8.872
−8.348
3.419
1.00
42.43
O

ATOM
1662
CB
LYS
X
220
6.678
−6.799
1.518
1.00
73.70
C

ATOM
1663
CG
LYS
X
220
5.259
−6.783
0.943
1.00
73.70
C

ATOM
1664
CD
LYS
X
220
4.838
−5.401
0.434
1.00
73.70
C

ATOM
1665
CE
LYS
X
220
3.562
−5.457
−0.439
1.00
73.70
C

ATOM
1666
NZ
LYS
X
220
2.275
−5.735
0.293
1.00
73.70
N

ATOM
1667
N
LYS
X
221
9.615
−8.240
1.321
1.00
58.48
N

ATOM
1668
CA
LYS
X
221
11.011
−8.294
1.725
1.00
58.48
C

ATOM
1669
C
LYS
X
221
11.868
−7.358
0.896
1.00
58.48
C

ATOM
1670
O
LYS
X
221
11.467
−6.915
−0.174
1.00
58.48
O

ATOM
1671
CB
LYS
X
221
11.544
−9.715
1.571
1.00
62.60
C

ATOM
1672
CG
LYS
X
221
10.755
−10.746
2.336
1.00
62.60
C

ATOM
1673
CD
LYS
X
221
11.248
−12.142
2.017
1.00
62.60
C

ATOM
1674
CE
LYS
X
221
10.363
−13.186
2.679
1.00
62.60
C

ATOM
1675
NZ
LYS
X
221
10.650
−14.566
2.186
1.00
62.60
N

ATOM
1676
N
VAL
X
222
13.057
−7.064
1.403
1.00
72.80
N

ATOM
1677
CA
VAL
X
222
13.992
−6.195
0.711
1.00
72.80
C

ATOM
1678
C
VAL
X
222
15.401
−6.640
1.027
1.00
72.80
C

ATOM
1679
O
VAL
X
222
15.625
−7.394
1.973
1.00
72.80
O

ATOM
1680
CB
VAL
X
222
13.836
−4.737
1.139
1.00
49.89
C

ATOM
1681
CG1
VAL
X
222
12.594
−4.154
0.507
1.00
49.89
C

ATOM
1682
CG2
VAL
X
222
13.770
−4.647
2.660
1.00
49.89
C

ATOM
1683
N
ILE
X
223
16.353
−6.163
0.235
1.00
61.92
N

ATOM
1684
CA
ILE
X
223
17.736
−6.535
0.430
1.00
61.92
C

ATOM
1685
C
ILE
X
223
18.686
−5.360
0.275
1.00
61.92
C

ATOM
1686
O
ILE
X
223
18.678
−4.659
−0.743
1.00
61.92
O

ATOM
1687
CB
ILE
X
223
18.126
−7.648
−0.551
1.00
57.10
C

ATOM
1688
CG1
ILE
X
223
17.293
−8.895
−0.252
1.00
57.10
C

ATOM
1689
CG2
ILE
X
223
19.607
−7.962
−0.438
1.00
57.10
C

ATOM
1690
CD1
ILE
X
223
17.449
−10.002
−1.266
1.00
57.10
C

ATOM
1691
N
ALA
X
224
19.492
−5.152
1.314
1.00
67.06
N

ATOM
1692
CA
ALA
X
224
20.492
−4.095
1.346
1.00
67.06
C

ATOM
1693
C
ALA
X
224
21.816
−4.847
1.277
1.00
67.06
C

ATOM
1694
O
ALA
X
224
22.138
−5.598
2.191
1.00
67.06
O

ATOM
1695
CB
ALA
X
224
20.379
−3.338
2.637
1.00
24.94
C

ATOM
1696
N
LYS
X
225
22.582
−4.658
0.206
1.00
60.46
N

ATOM
1697
CA
LYS
X
225
23.830
−5.399
0.054
1.00
60.46
C

ATOM
1698
C
LYS
X
225
24.886
−5.140
1.111
1.00
60.46
C

ATOM
1699
O
LYS
X
225
25.169
−6.009
1.936
1.00
60.46
O

ATOM
1700
CB
LYS
X
225
24.439
−5.163
−1.331
1.00
74.44
C

ATOM
1701
CG
LYS
X
225
25.649
−6.077
−1.656
1.00
74.44
C

ATOM
1702
CD
LYS
X
225
26.948
−5.655
−0.947
1.00
74.44
C

ATOM
1703
CE
LYS
X
225
28.105
−6.602
−1.248
1.00
74.44
C

ATOM
1704
NZ
LYS
X
225
27.919
−7.947
−0.630
1.00
74.44
N

ATOM
1705
N
ASP
X
226
25.486
−3.955
1.073
1.00
73.47
N

ATOM
1706
CA
ASP
X
226
26.532
−3.608
2.024
1.00
73.47
C

ATOM
1707
C
ASP
X
226
25.971
−3.093
3.346
1.00
73.47
C

ATOM
1708
O
ASP
X
226
25.731
−3.871
4.268
1.00
73.47
O

ATOM
1709
CB
ASP
X
226
27.454
−2.556
1.413
1.00
99.00
C

ATOM
1710
CG
ASP
X
226
26.739
−1.248
1.143
1.00
99.00
C

ATOM
1711
OD1
ASP
X
226
25.656
−1.286
0.520
1.00
99.00
O

ATOM
1712
OD2
ASP
X
226
27.254
−0.186
1.559
1.00
99.00
O

ATOM
1713
N
VAL
X
227
25.756
−1.782
3.426
1.00
69.61
N

ATOM
1714
CA
VAL
X
227
25.248
−1.131
4.625
1.00
69.61
C

ATOM
1715
C
VAL
X
227
25.956
−1.600
5.892
1.00
69.61
C

ATOM
1716
O
VAL
X
227
25.578
−1.239
7.008
1.00
69.61
O

ATOM
1717
CB
VAL
X
227
23.724
−1.323
4.777
1.00
47.31
C

ATOM
1718
CG1
VAL
X
227
23.017
−0.455
3.767
1.00
47.31
C

ATOM
1719
CG2
VAL
X
227
23.333
−2.789
4.607
1.00
47.31
C

ATOM
1720
N
ILE
X
228
26.996
−2.402
5.696
1.00
59.48
N

ATOM
1721
CA
ILE
X
228
27.812
−2.945
6.771
1.00
59.48
C

ATOM
1722
C
ILE
X
228
29.143
−3.325
6.131
1.00
59.48
C

ATOM
1723
O
ILE
X
228
29.323
−4.446
5.652
1.00
59.48
O

ATOM
1724
CB
ILE
X
228
27.186
−4.202
7.391
1.00
91.02
C

ATOM
1725
CG1
ILE
X
228
25.775
−3.900
7.893
1.00
91.02
C

ATOM
1726
CG2
ILE
X
228
28.047
−4.678
8.547
1.00
91.02
C

ATOM
1727
CD1
ILE
X
228
25.093
−5.087
8.526
1.00
91.02
C

ATOM
1728
N
PRO
X
229
30.088
−2.380
6.102
1.00
94.81
N

ATOM
1729
CA
PRO
X
229
31.418
−2.574
5.523
1.00
94.81
C

ATOM
1730
C
PRO
X
229
32.168
−3.765
6.109
1.00
94.81
C

ATOM
1731
O
PRO
X
229
32.286
−4.815
5.479
1.00
94.81
O

ATOM
1732
CB
PRO
X
229
32.112
−1.249
5.823
1.00
54.91
C

ATOM
1733
CG
PRO
X
229
30.978
−0.268
5.818
1.00
54.91
C

ATOM
1734
CD
PRO
X
229
29.934
−1.005
6.605
1.00
54.91
C

ATOM
1735
N
ALA
X
230
32.672
−3.574
7.323
1.00
99.00
N

ATOM
1736
CA
ALA
X
230
33.433
−4.576
8.067
1.00
99.00
C

ATOM
1737
C
ALA
X
230
34.068
−3.764
9.179
1.00
99.00
C

ATOM
1738
O
ALA
X
230
34.157
−4.199
10.329
1.00
99.00
O

ATOM
1739
CB
ALA
X
230
34.511
−5.199
7.189
1.00
44.67
C

ATOM
1740
N
ASN
X
231
34.501
−2.564
8.806
1.00
99.00
N

ATOM
1741
CA
ASN
X
231
35.111
−1.626
9.735
1.00
99.00
C

ATOM
1742
C
ASN
X
231
33.962
−0.974
10.493
1.00
99.00
C

ATOM
1743
O
ASN
X
231
34.052
0.173
10.931
1.00
99.00
O

ATOM
1744
CB
ASN
X
231
35.909
−0.561
8.974
1.00
99.00
C

ATOM
1745
CG
ASN
X
231
35.043
0.248
8.020
1.00
99.00
C

ATOM
1746
OD1
ASN
X
231
34.447
−0.296
7.092
1.00
99.00
O

ATOM
1747
ND2
ASN
X
231
34.969
1.554
8.249
1.00
99.00
N

ATOM
1748
N
TRP
X
232
32.871
−1.717
10.628
1.00
65.25
N

ATOM
1749
CA
TRP
X
232
31.705
−1.223
11.331
1.00
65.25
C

ATOM
1750
C
TRP
X
232
31.989
−1.144
12.823
1.00
65.25
C

ATOM
1751
O
TRP
X
232
32.555
−2.067
13.408
1.00
65.25
O

ATOM
1752
CB
TRP
X
232
30.533
−2.156
11.101
1.00
61.76
C

ATOM
1753
CG
TRP
X
232
30.747
−3.504
11.691
1.00
61.76
C

ATOM
1754
CD1
TRP
X
232
31.242
−4.607
11.063
1.00
61.76
C

ATOM
1755
CD2
TRP
X
232
30.461
−3.898
13.037
1.00
61.76
C

ATOM
1756
NE1
TRP
X
232
31.275
−5.671
11.934
1.00
61.76
N

ATOM
1757
CE2
TRP
X
232
30.802
−5.262
13.153
1.00
61.76
C

ATOM
1758
CE3
TRP
X
232
29.948
−3.229
14.155
1.00
61.76
C

ATOM
1759
CZ2
TRP
X
232
30.645
−5.975
14.346
1.00
61.76
C

ATOM
1760
CZ3
TRP
X
232
29.792
−3.934
15.338
1.00
61.76
C

ATOM
1761
CH2
TRP
X
232
30.140
−5.296
15.425
1.00
61.76
C

ATOM
1762
N
ARG
X
233
31.583
−0.045
13.440
1.00
68.52
N

ATOM
1763
CA
ARG
X
233
31.802
0.148
14.864
1.00
68.52
C

ATOM
1764
C
ARG
X
233
30.484
0.471
15.551
1.00
68.52
C

ATOM
1765
O
ARG
X
233
29.679
1.242
15.032
1.00
68.52
O

ATOM
1766
CB
ARG
X
233
32.792
1.287
15.082
1.00
98.84
C

ATOM
1767
CG
ARG
X
233
32.353
2.602
14.465
1.00
98.84
C

ATOM
1768
CD
ARG
X
233
33.383
3.682
14.721
1.00
98.84
C

ATOM
1769
NE
ARG
X
233
32.923
4.997
14.288
1.00
98.84
N

ATOM
1770
CZ
ARG
X
233
33.654
6.103
14.368
1.00
98.84
C

ATOM
1771
NH1
ARG
X
233
34.883
6.054
14.865
1.00
98.84
N

ATOM
1772
NH2
ARG
X
233
33.157
7.259
13.952
1.00
98.84
N

ATOM
1773
N
PRO
X
234
30.247
−0.110
16.738
1.00
70.12
N

ATOM
1774
CA
PRO
X
234
28.992
0.164
17.440
1.00
70.12
C

ATOM
1775
C
PRO
X
234
28.644
1.644
17.406
1.00
70.12
C

ATOM
1776
O
PRO
X
234
29.528
2.496
17.428
1.00
70.12
O

ATOM
1777
CB
PRO
X
234
29.270
−0.348
18.847
1.00
75.00
C

ATOM
1778
CG
PRO
X
234
30.156
−1.524
18.582
1.00
75.00
C

ATOM
1779
CD
PRO
X
234
31.126
−0.966
17.554
1.00
75.00
C

ATOM
1780
N
ASP
X
235
27.350
1.929
17.334
1.00
99.00
N

ATOM
1781
CA
ASP
X
235
26.842
3.292
17.285
1.00
99.00
C

ATOM
1782
C
ASP
X
235
26.915
3.837
15.855
1.00
99.00
C

ATOM
1783
O
ASP
X
235
26.174
4.755
15.495
1.00
99.00
O

ATOM
1784
CB
ASP
X
235
27.632
4.191
18.245
1.00
99.00
C

ATOM
1785
CG
ASP
X
235
26.953
5.527
18.495
1.00
99.00
C

ATOM
1786
OD1
ASP
X
235
26.740
6.286
17.524
1.00
99.00
O

ATOM
1787
OD2
ASP
X
235
26.632
5.820
19.667
1.00
99.00
O

ATOM
1788
N
ALA
X
236
27.797
3.265
15.037
1.00
67.41
N

ATOM
1789
CA
ALA
X
236
27.935
3.711
13.655
1.00
67.41
C

ATOM
1790
C
ALA
X
236
26.593
3.570
12.943
1.00
67.41
C

ATOM
1791
O
ALA
X
236
25.871
2.594
13.158
1.00
67.41
O

ATOM
1792
CB
ALA
X
236
28.993
2.895
12.942
1.00
42.14
C

ATOM
1793
N
VAL
X
237
26.266
4.545
12.097
1.00
99.00
N

ATOM
1794
CA
VAL
X
237
24.999
4.535
11.369
1.00
99.00
C

ATOM
1795
C
VAL
X
237
25.158
4.366
9.862
1.00
99.00
C

ATOM
1796
O
VAL
X
237
25.962
5.048
9.229
1.00
99.00
O

ATOM
1797
CB
VAL
X
237
24.211
5.830
11.618
1.00
54.48
C

ATOM
1798
CG1
VAL
X
237
22.843
5.733
10.981
1.00
54.48
C

ATOM
1799
CG2
VAL
X
237
24.081
6.075
13.098
1.00
54.48
C

ATOM
1800
N
TYR
X
238
24.369
3.462
9.292
1.00
46.71
N

ATOM
1801
CA
TYR
X
238
24.417
3.194
7.860
1.00
46.71
C

ATOM
1802
C
TYR
X
238
23.058
3.423
7.224
1.00
46.71
C

ATOM
1803
O
TYR
X
238
22.033
3.006
7.749
1.00
46.71
O

ATOM
1804
CB
TYR
X
238
24.868
1.756
7.619
1.00
70.02
C

ATOM
1805
CG
TYR
X
238
26.169
1.445
8.313
1.00
70.02
C

ATOM
1806
CD1
TYR
X
238
27.392
1.696
7.694
1.00
70.02
C

ATOM
1807
CD2
TYR
X
238
26.179
0.952
9.616
1.00
70.02
C

ATOM
1808
CE1
TYR
X
238
28.595
1.464
8.354
1.00
70.02
C

ATOM
1809
CE2
TYR
X
238
27.371
0.718
10.289
1.00
70.02
C

ATOM
1810
CZ
TYR
X
238
28.577
0.975
9.653
1.00
70.02
C

ATOM
1811
OH
TYR
X
238
29.760
0.739
10.323
1.00
70.02
O

ATOM
1812
N
THR
X
239
23.061
4.109
6.092
1.00
60.41
N

ATOM
1813
CA
THR
X
239
21.833
4.383
5.370
1.00
60.41
C

ATOM
1814
C
THR
X
239
21.829
3.569
4.078
1.00
60.41
C

ATOM
1815
O
THR
X
239
22.831
3.508
3.369
1.00
60.41
O

ATOM
1816
CB
THR
X
239
21.717
5.879
5.060
1.00
82.69
C

ATOM
1817
OG1
THR
X
239
20.762
6.083
4.012
1.00
82.69
O

ATOM
1818
CG2
THR
X
239
23.067
6.437
4.658
1.00
82.69
C

ATOM
1819
N
SER
X
240
20.700
2.935
3.784
1.00
50.19
N

ATOM
1820
CA
SER
X
240
20.569
2.105
2.590
1.00
50.19
C

ATOM
1821
C
SER
X
240
19.832
2.801
1.460
1.00
50.19
C

ATOM
1822
O
SER
X
240
19.341
3.910
1.606
1.00
50.19
O

ATOM
1823
CB
SER
X
240
19.794
0.843
2.925
1.00
47.09
C

ATOM
1824
OG
SER
X
240
18.447
1.184
3.214
1.00
47.09
O

ATOM
1825
N
ASN
X
241
19.762
2.115
0.328
1.00
62.17
N

ATOM
1826
CA
ASN
X
241
19.053
2.591
−0.846
1.00
62.17
C

ATOM
1827
C
ASN
X
241
18.068
1.464
−1.088
1.00
62.17
C

ATOM
1828
O
ASN
X
241
18.227
0.663
−2.001
1.00
62.17
O

ATOM
1829
CB
ASN
X
241
20.010
2.735
−2.023
1.00
98.83
C

ATOM
1830
CG
ASN
X
241
20.975
1.575
−2.123
1.00
98.83
C

ATOM
1831
OD1
ASN
X
241
20.568
0.424
−2.272
1.00
98.83
O

ATOM
1832
ND2
ASN
X
241
22.267
1.872
−2.034
1.00
98.83
N

ATOM
1833
N
VAL
X
242
17.059
1.404
−0.229
1.00
51.38
N

ATOM
1834
CA
VAL
X
242
16.034
0.374
−0.269
1.00
51.38
C

ATOM
1835
C
VAL
X
242
14.926
0.852
0.655
1.00
51.38
C

ATOM
1836
O
VAL
X
242
15.202
1.466
1.678
1.00
51.38
O

ATOM
1837
CB
VAL
X
242
16.602
−0.968
0.255
1.00
61.13
C

ATOM
1838
CG1
VAL
X
242
15.549
−1.721
1.061
1.00
61.13
C

ATOM
1839
CG2
VAL
X
242
17.090
−1.815
−0.914
1.00
61.13
C

ATOM
1840
N
GLN
X
243
13.677
0.575
0.313
1.00
34.48
N

ATOM
1841
CA
GLN
X
243
12.571
1.025
1.149
1.00
34.48
C

ATOM
1842
C
GLN
X
243
11.300
0.280
0.758
1.00
34.48
C

ATOM
1843
O
GLN
X
243
11.295
−0.443
−0.229
1.00
34.48
O

ATOM
1844
CB
GLN
X
243
12.394
2.539
0.984
1.00
31.17
C

ATOM
1845
CG
GLN
X
243
11.115
3.083
1.553
1.00
31.17
C

ATOM
1846
CD
GLN
X
243
11.088
3.108
3.067
1.00
31.17
C

ATOM
1847
OE1
GLN
X
243
11.813
3.875
3.685
1.00
31.17
O

ATOM
1848
NE2
GLN
X
243
10.246
2.283
3.668
1.00
31.17
N

ATOM
1849
N
PHE
X
244
10.236
0.427
1.537
1.00
42.15
N

ATOM
1850
CA
PHE
X
244
8.980
−0.237
1.229
1.00
42.15
C

ATOM
1851
C
PHE
X
244
8.001
0.746
0.600
1.00
42.15
C

ATOM
1852
O
PHE
X
244
7.683
1.792
1.174
1.00
42.15
O

ATOM
1853
CB
PHE
X
244
8.344
−0.833
2.485
1.00
37.58
C

ATOM
1854
CG
PHE
X
244
9.016
−2.080
2.982
1.00
37.58
C

ATOM
1855
CD1
PHE
X
244
10.165
−2.010
3.760
1.00
37.58
C

ATOM
1856
CD2
PHE
X
244
8.503
−3.330
2.668
1.00
37.58
C

ATOM
1857
CE1
PHE
X
244
10.790
−3.169
4.216
1.00
37.58
C

ATOM
1858
CE2
PHE
X
244
9.120
−4.491
3.118
1.00
37.58
C

ATOM
1859
CZ
PHE
X
244
10.264
−4.411
3.892
1.00
37.58
C

ATOM
1860
N
TYR
X
245
7.511
0.409
−0.585
1.00
36.46
N

ATOM
1861
CA
TYR
X
245
6.570
1.281
−1.260
1.00
36.46
C

ATOM
1862
C
TYR
X
245
5.168
0.740
−1.163
1.00
36.46
C

ATOM
1863
O
TYR
X
245
4.224
1.551
−1.177
1.00
36.46
O

ATOM
1864
CB
TYR
X
245
7.000
1.446
−2.698
1.00
35.82
C

ATOM
1865
CG
TYR
X
245
8.261
2.249
−2.803
1.00
35.82
C

ATOM
1866
CD1
TYR
X
245
8.212
3.632
−2.903
1.00
35.82
C

ATOM
1867
CD2
TYR
X
245
9.496
1.634
−2.778
1.00
35.82
C

ATOM
1868
CE1
TYR
X
245
9.357
4.388
−2.981
1.00
35.82
C

ATOM
1869
CE2
TYR
X
245
10.659
2.379
−2.854
1.00
35.82
C

ATOM
1870
CZ
TYR
X
245
10.584
3.763
−2.958
1.00
35.82
C

ATOM
1871
OH
TYR
X
245
11.735
4.521
−3.050
1.00
35.82
O

ATOM
1872
OXT
TYR
X
245
5.050
−0.497
−1.065
1.00
35.82
O

TER
1873

TYR
X
245

HETATM
1874
C1
NAG
A
1
−1.681
−10.997
19.292
1.00
56.00
C

HETATM
1875
C2
NAG
A
1
−1.683
−12.434
18.767
1.00
56.00
C

HETATM
1876
C3
NAG
A
1
−0.703
−13.347
19.525
1.00
56.00
C

HETATM
1877
C4
NAG
A
1
0.560
−12.674
20.069
1.00
56.00
C

HETATM
1878
C5
NAG
A
1
0.445
−11.170
20.325
1.00
56.00
C

HETATM
1879
C6
NAG
A
1
1.802
−10.486
20.363
1.00
56.00
C

HETATM
1880
C7
NAG
A
1
−3.344
−14.077
18.220
1.00
56.00
C

HETATM
1881
C8
NAG
A
1
−4.366
−14.981
18.878
1.00
56.00
C

HETATM
1882
N2
NAG
A
1
−3.022
−12.975
18.880
1.00
56.00
N

HETATM
1883
O3
NAG
A
1
−0.278
−14.394
18.666
1.00
56.00
O

HETATM
1884
O4
NAG
A
1
0.902
−13.309
21.313
1.00
56.00
O

HETATM
1885
O5
NAG
A
1
−0.334
−10.527
19.298
1.00
56.00
O

HETATM
1886
O6
NAG
A
1
2.829
−11.363
19.856
1.00
56.00
O

HETATM
1887
O7
NAG
A
1
−2.847
−14.380
17.132
1.00
56.00
O

HETATM
1888
C1
NAG
A
2
2.191
−13.761
21.447
1.00
55.26
C

HETATM
1889
C2
NAG
A
2
2.393
−14.216
22.902
1.00
55.26
C

HETATM
1890
C3
NAG
A
2
3.683
−15.060
23.081
1.00
55.26
C

HETATM
1891
C4
NAG
A
2
3.738
−16.138
21.974
1.00
55.26
C

HETATM
1892
C5
NAG
A
2
3.626
−15.476
20.608
1.00
55.26
C

HETATM
1893
C6
NAG
A
2
3.683
−16.491
19.482
1.00
55.26
C

HETATM
1894
C7
NAG
A
2
1.524
−12.759
24.641
1.00
55.26
C

HETATM
1895
C8
NAG
A
2
0.071
−13.061
24.309
1.00
55.26
C

HETATM
1896
N2
NAG
A
2
2.461
−13.026
23.732
1.00
55.26
N

HETATM
1897
O3
NAG
A
2
3.667
−15.679
24.368
1.00
55.26
O

HETATM
1898
O4
NAG
A
2
4.927
−16.957
22.011
1.00
55.26
O

HETATM
1899
O5
NAG
A
2
2.351
−14.831
20.512
1.00
55.26
O

HETATM
1900
O6
NAG
A
2
4.926
−16.440
18.810
1.00
55.26
O

HETATM
1901
O7
NAG
A
2
1.797
−12.257
25.727
1.00
55.26
O

HETATM
1902
C1
MAN
A
3
5.898
−16.746
22.959
1.00
61.20
C

HETATM
1903
C2
MAN
A
3
7.261
−17.190
22.393
1.00
61.20
C

HETATM
1904
C3
MAN
A
3
7.457
−18.720
22.360
1.00
61.20
C

HETATM
1905
C4
MAN
A
3
6.971
−19.365
23.647
1.00
61.20
C

HETATM
1906
C5
MAN
A
3
5.541
−18.915
23.923
1.00
61.20
C

HETATM
1907
C6
MAN
A
3
5.004
−19.556
25.180
1.00
61.20
C

HETATM
1908
O2
MAN
A
3
8.331
−16.550
23.106
1.00
61.20
O

HETATM
1909
O3
MAN
A
3
8.855
−19.047
22.160
1.00
61.20
O

HETATM
1910
O4
MAN
A
3
7.033
−20.776
23.535
1.00
61.20
O

HETATM
1911
O5
MAN
A
3
5.530
−17.484
24.130
1.00
61.20
O

HETATM
1912
O6
MAN
A
3
6.066
−19.823
26.077
1.00
61.20
O

HETATM
1913
C1
MAN
A
4
9.104
−20.270
21.508
1.00
77.31
C

HETATM
1914
C2
MAN
A
4
10.609
−20.481
21.314
1.00
77.31
C

HETATM
1915
C3
MAN
A
4
11.163
−19.499
20.305
1.00
77.31
C

HETATM
1916
C4
MAN
A
4
10.406
−19.704
18.986
1.00
77.31
C

HETATM
1917
C5
MAN
A
4
8.929
−19.441
19.251
1.00
77.31
C

HETATM
1918
C6
MAN
A
4
8.043
−19.588
18.040
1.00
77.31
C

HETATM
1919
O2
MAN
A
4
10.817
−21.782
20.798
1.00
77.31
O

HETATM
1920
O3
MAN
A
4
12.550
−19.756
20.127
1.00
77.31
O

HETATM
1921
O4
MAN
A
4
10.893
−18.814
17.992
1.00
77.31
O

HETATM
1922
O5
MAN
A
4
8.444
−20.365
20.248
1.00
77.31
O

HETATM
1923
O6
MAN
A
4
6.676
−19.417
18.401
1.00
77.31
O

HETATM
1924
C1
XYS
A
6
8.653
−15.288
22.495
1.00
55.63
C

HETATM
1925
C2
XYS
A
6
9.694
−15.434
21.383
1.00
55.63
C

HETATM
1926
C3
XYS
A
6
11.060
−14.827
21.679
1.00
55.63
C

HETATM
1927
C4
XYS
A
6
10.882
−13.425
22.289
1.00
55.63
C

HETATM
1928
C5
XYS
A
6
10.206
−13.683
23.667
1.00
55.63
C

HETATM
1929
O2
XYS
A
6
10.016
−16.823
21.218
1.00
55.63
O

HETATM
1930
O3
XYS
A
6
11.722
−14.753
20.373
1.00
55.63
O

HETATM
1931
O4
XYS
A
6
12.180
−12.880
22.548
1.00
55.63
O

HETATM
1932
O5
XYS
A
6
8.852
−14.240
23.500
1.00
55.63
O

HETATM
1933
C1
MAN
A
5
5.864
−20.604
27.410
1.00
75.97
C

HETATM
1934
C2
MAN
A
5
5.121
−19.336
27.902
1.00
75.97
C

HETATM
1935
C3
MAN
A
5
6.010
−18.068
27.970
1.00
75.97
C

HETATM
1936
C4
MAN
A
5
7.422
−18.366
28.475
1.00
75.97
C

HETATM
1937
C5
MAN
A
5
7.992
−19.582
27.747
1.00
75.97
C

HETATM
1938
C6
MAN
A
5
9.400
−19.942
28.166
1.00
75.97
C

HETATM
1939
O2
MAN
A
5
4.525
−19.580
29.163
1.00
75.97
O

HETATM
1940
O3
MAN
A
5
5.406
−17.116
28.838
1.00
75.97
O

HETATM
1941
O4
MAN
A
5
8.248
−17.234
28.248
1.00
75.97
O

HETATM
1942
O5
MAN
A
5
7.162
−20.729
28.007
1.00
75.97
O

HETATM
1943
O6
MAN
A
5
10.361
−19.272
27.362
1.00
75.97
O

HETATM
1944
O
HOH

1
11.188
8.002
−3.637
1.00
57.41
O

HETATM
1945
O
HOH

2
−0.416
−10.450
15.145
1.00
57.41
O

HETATM
1946
O
HOH

3
−0.492
−9.251
12.441
1.00
57.41
O

HETATM
1947
O
HOH

4
−7.200
−4.855
27.320
1.00
57.41
O

HETATM
1948
O
HOH

5
1.904
−3.678
3.473
1.00
57.41
O

HETATM
1949
O
HOH

6
−0.543
−16.030
16.418
1.00
57.41
O

HETATM
1950
O
HOH

7
−17.147
0.652
5.613
1.00
57.41
O

HETATM
1951
O
HOH

8
−16.533
10.658
9.845
1.00
57.41
O

HETATM
1952
O
HOH

9
17.841
−14.159
−2.339
1.00
57.41
O

HETATM
1953
O
HOH

10
−13.522
−1.089
9.451
1.00
63.85
O

HETATM
1954
O
HOH

11
−16.163
21.987
15.522
1.00
66.61
O

HETATM
1955
O
HOH

12
25.884
−21.759
9.015
1.00
42.35
O

HETATM
1956
O
HOH

13
10.841
17.340
13.889
1.00
79.50
O

HETATM
1957
O
HOH

14
−0.184
13.888
5.045
1.00
44.33
O

HETATM
1958
O
HOH

15
9.810
−20.435
24.512
1.00
75.26
O

HETATM
1959
O
HOH

16
−2.242
−0.783
28.049
1.00
45.90
O

HETATM
1960
O
HOH

17
−1.601
−15.062
12.117
1.00
46.17
O

HETATM
1961
C1
FCA
A
7
−0.851
−15.400
19.952
1.00
88.23
C

HETATM
1962
C2
FCA
A
7
−1.198
−15.522
21.457
1.00
88.23
C

HETATM
1963
C3
FCA
A
7
−1.707
−16.987
21.710
1.00
88.23
C

HETATM
1964
C4
FCA
A
7
−0.606
−18.028
21.285
1.00
88.23
C

HETATM
1965
C5
FCA
A
7
−0.267
−17.763
19.785
1.00
88.23
C

HETATM
1966
C6
FCA
A
7
0.811
−18.713
19.204
1.00
88.23
C

HETATM
1967
O2
FCA
A
7
−2.202
−14.574
21.849
1.00
88.23
O

HETATM
1968
O3
FCA
A
7
−2.050
−17.184
23.084
1.00
88.23
O

HETATM
1969
O4
FCA
A
7
0.575
−17.816
22.092
1.00
88.23
O

HETATM
1970
O5
FCA
A
7
0.171
−16.377
19.623
1.00
88.23
O

CONECT
35
36
41

CONECT
36
35
37
39

CONECT
37
36
38

CONECT
38
37

CONECT
39
36
40

CONECT
40
39
41
42

CONECT
41
35
40

CONECT
42
40

CONECT
50
1874

CONECT
284
483

CONECT
483
284

CONECT
499
1034

CONECT
544
593

CONECT
593
544

CONECT
1034
499

CONECT
1874
50
1875
1885

CONECT
1875
1874
1876
1882

CONECT
1876
1875
1877
1883

CONECT
1877
1876
1878
1884

CONECT
1878
1877
1879
1885

CONECT
1879
1878
1886

CONECT
1880
1881
1882
1887

CONECT
1881
1880

CONECT
1882
1875
1880

CONECT
1883
1876
1961

CONECT
1884
1877
1888

CONECT
1885
1874
1878

CONECT
1886
1879

CONECT
1887
1880

CONECT
1888
1884
1889
1899

CONECT
1889
1888
1890
1896

CONECT
1890
1889
1891
1897

CONECT
1891
1890
1892
1898

CONECT
1892
1891
1893
1899

CONECT
1893
1892
1900

CONECT
1894
1895
1896
1901

CONECT
1895
1894

CONECT
1896
1889
1894

CONECT
1897
1890

CONECT
1898
1891
1902

CONECT
1899
1888
1892

CONECT
1900
1893

CONECT
1901
1894

CONECT
1902
1898
1903
1911

CONECT
1903
1902
1904
1908

CONECT
1904
1903
1905
1909

CONECT
1905
1904
1906
1910

CONECT
1906
1905
1907
1911

CONECT
1907
1906
1912

CONECT
1908
1903
1924

CONECT
1909
1904
1913

CONECT
1910
1905

CONECT
1911
1902
1906

CONECT
1912
1907
1933

CONECT
1913
1909
1914
1922

CONECT
1914
1913
1915
1919

CONECT
1915
1914
1916
1920

CONECT
1916
1915
1917
1921

CONECT
1917
1916
1918
1922

CONECT
1918
1917
1923

CONECT
1919
1914

CONECT
1920
1915

CONECT
1921
1916

CONECT
1922
1913
1917

CONECT
1923
1918

CONECT
1924
1908
1925
1932

CONECT
1925
1924
1926
1929

CONECT
1926
1925
1927
1930

CONECT
1927
1926
1928
1931

CONECT
1928
1927
1932

CONECT
1929
1925

CONECT
1930
1926

CONECT
1931
1927

CONECT
1932
1924
1928

CONECT
1933
1912
1934
1942

CONECT
1934
1933
1935
1939

CONECT
1935
1934
1936
1940

CONECT
1936
1935
1937
1941

CONECT
1937
1936
1938
1942

CONECT
1938
1937
1943

CONECT
1939
1934

CONECT
1940
1935

CONECT
1941
1936

CONECT
1942
1933
1937

CONECT
1943
1938

CONECT
1961
1883
1962
1970

CONECT
1962
1961
1963
1967

CONECT
1963
1962
1964
1968

CONECT
1964
1963
1965
1969

CONECT
1965
1964
1966
1970

CONECT
1966
1965

CONECT
1967
1962

CONECT
1968
1963

CONECT
1969
1964

CONECT
1970
1961
1965

MASTER 320 0 8 4 15 0 0 6 1969 1 95 19

END

TABLE 2

Refinement statistics and final model statistics for EXPB1

Resolution, Å
2.75

Cell

a, Å
113.408

b, Å
44.512

c, Å
69.467

β, °
124.64

Space group
C 2

Final R_free, %
29.07

Final R, %
23.32

No. of reflections
6,640 (working)/367(test)

No. of atoms
1,969

B(iso) 2 of protein atoms, Å
55.5

Main chain
53.4

Side chain
57.6

rmsd in bond lengths, Å
0.008

rmsd in bond angles, °
1.68

Solvent content, %
53.9

Ramachandran plot, %

Most favored region, %
91.9

Generously allowed, %
6.6

Supporting Text

Structure Solution and Refinement. After several cycles of rigid body refinement the maps still looked noisy. To improve this, density modification was performed by using the program CNS (1). Solvent density modification and density truncation features were used. The resulting maps gradually helped in modeling regions of the missing N-terminal residues and the loop between residues 29 and 38. The side chains that were different in Phl p 1 compared to EXPB1 could also be corrected, and the four extra residues at the C terminus could be located. The polysaccharide covalently linked to Asn-10 was modeled as shown in FIG. 2. Only group β-factor refinement was used, given that the resolution of the data was 2.75 Å. After several iterations of modeling using the program 0 (2) and density modification and refinement using the program CNS, the R-factors converged at 23.32% and 29.07%, respectively. At the very end of the refinement a total of nine water molecules could be located. As reported by PROCHECK (3), all residues lie in or close to the allowed regions of the Ramachandran plot. The first three residues at the N terminus are disordered and are not part of the model.

Comparison with Phl p 1 (PDB ID code 1N10). Compared to the 2.9-Å structure of 1N10, which has a dimer in the asymmetric unit, EXPB1, with a monomer in the asymmetric unit, is solved to a better resolution (2.75 Å). The loop consisting of residues 29-38 is not resolved in 1N10 but has good electron density in EXPB1. This is an important loop because it contains D37, a potential candidate for the catalytic base. The first 15 residues at the N-terminal extension are oriented entirely differently in the two structures (leading to successful molecular replacement when omitted). The N-terminal strand in 1N10 extends out away from the protein and interacts with a second monomer. Because the recombinantly produced Phl p 1 used to solve the 1N10 structure was not native protein, the processing of the N-terminal extension appears to be atypical (the hydroxylation of prolines is lacking, and glycosylation pattern at N10 is probably different and was resolved to only one GlcNac).

When the Cα carbon atoms of both D1 and D2 are superimposed for the two proteins, the rmsd is 1.84 Å. Superposition of the D1s alone (excluding the first 15 residues at the N terminus) reveals a good overlap (rmsd of 0.88 Å) whereas the D2s overlap poorly (rmsd of 1.82 Å). Comparison of the overlapping structures shows that the Cα of W194 (D2), which is a crucial part of the putative binding groove, is displaced by 4 Å in 1N10 and its side chain is rotated and displaced by almost 12 Å. However, the tryptophan ring continues to stay in the same plane as the other residues at the base of the groove, and hence a sugar could still bind in a fashion similar to that for EXPB1.

1. Brunger A T, Adams P D, Clore G M, DeLano W L, Gros P, Grosse-Kunstleve R W, Jiang J S, Kuszewski J, Nilges M, Pannu N S, et al. (1998) Acta Crystallogr D 54:905-921.
2. Jones T A, Zou J Y, Cowan S W, Kjeldgaard M (1991) Acta Crystallogr A 47:110-119.
3. Laskowski R A, Macarthur M W, Moss D S, Thornton J M (1993) J Appl Crystallogr 26:283-291.

Expansins have the many conserved domains as shown in FIG. 6. The signal peptide directs the nascent polypeptide into the ER/Golgi secretory pathway. This part of the protein (typically 22-25 amino acids) is removed as the protein enters the ER.

The mature protein is ˜25-27 kDa and consists of two domains, an amino-terminal domain of ˜120 amino acid residues (green in structure) with structural and sequence similarity to family-45 endoglucanases (EG45-like domain) and a carboxy-terminal domain of ˜98 amino acid residues (cyan in structure) that is hypothesized to function as a polysaccharide-binding domain (this is not experimentally established).

FIG. 4 shows is a crystal structure model (PDB code 2HCZ) of a beta-expansin in which the two domains are colored green and cyan. The two domains form a long, open planar surface with conserved polar residues that could hydrogen bond to a polysaccharide and four conserved aromatic residues (red below) that could bind the sugar rings by Van der Waals forces.

Increased activity and efficiency of expansin-like proteins

Information

Publication Number

Date Filed

Date Published

Inventors

Original Assignees

CPC

US Classifications

International Classifications

Abstract

Description

Claims

CROSS-REFERENCE TO RELATED APPLICATIONS

Provisional Applications (1)