WO2012137012A1

WO2012137012A1 - Crystal structure of an a2a adenosine receptor

Info

Publication number: WO2012137012A1
Application number: PCT/GB2012/050775
Authority: WO
Inventors: Guillaume Pierre Lebon; Christopher Gordon Tate
Original assignee: Heptares Therapeutics Ltd
Priority date: 2011-04-06
Filing date: 2012-04-05
Publication date: 2012-10-11

Abstract

The invention provides a method of predicting a three dimensional structural representation of a target protein of unknown structure, or part thereof, comprising : providing the coordinates of the adenosine A_2A receptor structure listed in Table (i), optionally varied by a root mean square deviation of residue backbone atoms of not more than 1.08 A, or selected coordinates thereof; and predicting the three-dimensional structural representation of the target protein, or part thereof, by modelling the structural representation on all or the selected coordinates of the adenosine A_2A receptor. The invention also provides the use of the adenosine A_2A receptor coordinates to select or design one or more binding partners of adenosine A_2A receptor.

Description

CRYSTAL STRUCTURE OF AN A2A ADENOSINE RECEPTOR

The present invention relates to protein crystal structures and their use in identifying protein binding partners and in protein structure determination. In particular, it relates to the crystal structure of an adenosine Aaa, receptor and uses thereof.

The listing or discussion of an apparently prior-published document in this specification should not necessarily be taken as an acknowledgement that the document is part of the state of the art or is common general knowledge.

G protein-coupled receptors (GPCRs) are integral membrane proteins mediating the signalling of a diverse set of ligands including neurotransmitters and metabolites. In humans, there are approximately 370 non-sensory receptors, representing the site of action for ~30% of clinically used drugs. Activation of the receptor results in a conformational change propagated to the intracellular surface where the receptor interacts with heterotrimeric G proteins to regulate signalling to ion channels and enzyme pathways. GPCRs can also signal independently of G proteins through β-arrestin and are known to exist as dimers. This multimeric state is likely central to their function and also delivery to the membrane, however, the molecular mechanism and details of GPCR oligomerization remains poorly understood.

The adenosine A2A receptor is one of 4 GPCRs (A-,, A2A, A_2B, A₃) activated by adenosine. Adenosine represents an important modulator of the central nervous system and periphery. In the brain adenosine controls neuronal excitability and the psychoactive effects of caffeine are mediated by adenosine receptors. A2A receptors are located in the striatum and are considered a target for neurodegenerative disease⁴⁶. A_¾ receptors are also expressed on the vasculature and immune cells where they have vasodilatory and anti-inflammatory effects ⁴⁷⁴⁸. There is growing evidence that drugs acting at adenosine receptors represent promising approaches in a wide range of diseases ⁸. Mechanistic understanding of ligand binding and activation, as well as our ability to design drugs for GPCRs is hampered by the lack of structural information. Structures of rhodopsin in conformationally distinct states have indicated that receptor activation results in an outward tilt of transmembrane helix (TM) 6 and a rearrangement of hydrogen bonding networks connecting the ligand binding pocket to the cytoplasmic face. The first non- rhodopsin GPCR structure to be obtained was the p₂-adrenergic receptor in complex with an antibody fragment bound to the third intracellular loop (ICL) 3 - a critical domain of the receptor which mediates G protein coupling A higher resolution structure of the p₂-AR was obtained by fusing T4 lysozyme into ICL3⁵⁵ and the same methodology was used to obtain the first structure of the adenosine A2A receptor (A2A- 4L)⁸. The conformation of these receptors remains unclear since insertion of the T4 lysozyme alters the pharmacology and prevents signalling. A structure has also been obtained for the adrenergic receptor ( iAR) using a mutagenesis approach to stabilise the antagonist state¹⁹. This was the first non- rhodopsin structure to clearly show features of the cytoplasmic regions of the receptor and revealed the presence of a short well defined helix in ICL2. However, in this structure ICL3 was truncated to assist in crystallisation.

The inventors have now solved the structure of the adenosine A2A receptor in complex with the agonist adenosine and the synthetic agonist NECA (5'-N-ethylcarboxamidoadenosine). The structure of the adenosine A2A receptor described here provides new insight into the structural features which define the GPCR active state, the regions which interact with signal transduction proteins, and how receptors interact to form signalling complexes.

The A2A receptor used in this study was A2A-GL31, is a truncated version of the A2A receptor containing residues 1 to 316, with the C-terminus (residues 317 to 412) removed. Four thermostabilising point mutations (L48A, A54L, T65A and Q89A) were introduced and the mutation N154A was finally added to removed the glycosylation site.

The coordinates of the adenosine A_¾ receptor can be utilised and manipulated in many different ways with wide ranging applications including the fitting of binding partners, homology modelling and structure solution, analysis of ligand interactions and drug discovery.

Accordingly, a first aspect of the invention provides a method of predicting a three dimensional structural representation of a target protein of unknown structure, or part thereof, comprising:

providing the coordinates of the human adenosine A2A receptor structure listed in Table (i), optionally varied by a root mean square deviation of residue backbone atoms of not more than 1.08 A, or selected coordinates thereof; and

predicting the three-dimensional structural representation of the target protein, or part thereof, by modelling the structural representation on all or the selected coordinates of the adenosine A2A receptor. Table (i) represents the parameters for modelling the C121 crystal system NECA-A₂AR-GL31 complex.

By a 'three dimensional structural representation' we include a computer generated representation or a physical representation. Typically, in all aspects of the invention which feature a structural representation, the representation is computer generated. Computer representations can be generated or displayed by commercially available software programs. Examples of software programs include but are not limited to QUANTA (Accelrys .COPYRIGHT.2001, 2002), O (Jones et al., Acta Crystallogr. A47, pp. 110-119 (1991)) and RIBBONS (Carson, J. Appl. Crystallogr., 24, pp. 9589-961 (1991)), which are incorporated herein by reference. Examples of representations include any of a wire-frame model, a chicken-wire model, a ball-and-stick model, a space-filling model, a stick model, a ribbon model, a snake model, an arrow and cylinder model, an electron density map or a molecular surface model. Certain software programs may also imbue these three dimensional representations with physico-chemical attributes which are known from the chemical composition of the molecule, such as residue charge, hydrophobicity, torsional and rotational degrees of freedom for the residue or segment, etc. Examples of software programs for calculating chemical energies are described below. Typically, the coordinates of the adenosine A_2A receptor structure used in the invention are those listed in Table (i). However, it is appreciated that it is not necessary to have recourse to the original coordinates listed in Table (i), and that any equivalent geometric representation derived from or obtained by reference to the original coordinates may be used.

Thus, for the avoidance of doubt, by 'the coordinates of the adenosine A₂A receptor structures listed in Table (ί)', we include any equivalent representation wherein the original coordinates have been reparameterised in some way. For example, the coordinates in Table (i) may undergo any mathematical transformation known in the art, such as a geometric transformation, and the resulting transformed coordinates can be used. For example, the coordinates of Table (i) may be transposed to a different origin and/or axes or may be rotated about an axis. Furthermore, it is possible to use the coordinates to calculate the psi and phi backbone torsion angles (as displayed on a Ramachandran plot) and the chi sidechain torsion angles for each residue in the protein. These angles together with the corresponding bond lengths, enable the construction of a geometric representation of the protein which may be used based on the parameters of psi, phi and chi angles and bond lengths. Thus, while the coordinates used are typically those in Table (i), the inventors recognise that any equivalent geometric representation of the adenosine A2A receptor structure, based on the coordinates listed in Table (i) may be used. Additionally, it is appreciated that changing the number and/or positions of the ligand molecule of the Tables does not generally affect the usefulness of the coordinates in the aspects of the invention. Thus, it is also within the scope of the invention if the number and/or positions of ligand molecules of the coordinates of Table (i) is varied. It will be appreciated that in all aspects of the invention which utilise the coordinates of the adenosine A2A receptor, it is not necessary to utilise all the coordinates of Table (i), but merely a portion of them, e.g. a set of coordinates representing atoms of particular interest in relation to a particular use. Such a portion of coordinates is referred to herein as 'selected coordinates'.

By 'selected coordinates', we include at least 5, 10 or 20 non-hydrogen protein atoms of the adenosine A_2A receptor structure, more preferably at least 50, 100, 200, 300, 400, 500, 600, 700, 800 or 900 atoms and even more preferably at least 1000, 1100, 1200, 1300, 1400, 1500, 1600, 1700, 1800, 1900, 2000, 2100 or 2200 non-hydrogen atoms. Preferably the selected coordinates pertain to at least 5, 10, 20 or 30 different amino acid residues (i.e. at least one atom from 5, 10, 20 or 30 different residues may be present), more preferably at least 40, 50, 60, 70, 80 or 90 residues, and even more preferably at least 100, 150, 200, 250 or 300 residues. Optionally, the selected coordinates may include one or more ligand atoms as set out in Table (i). Alternatively, the selected coordinates may exclude one or more atoms of the ligand.

In one example, the selected coordinates may comprise atoms of one or more amino acid residues that contribute to the adenosine binding site of the A2A receptor. For example, amino acid residues contributing to the adenosine binding site include amino acid residues Ile66, Ala63, lie 274, Val84, His278, Ser277, Leu249, Trp246, Thr88, Asn181 , His250, Met177, Asn253, Met 270, Glu169 and Phe168, according to the numbering of the adenosine A2A receptor sequence as set out in Figure 13. Thus the selected coordinates may comprise one or more atoms from any one or more (e.g. at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15 or 16 amino acid residues) of amino acid residues Ile66, Ala63, lie 274, Val84, His278, Ser277, Leu249, Trp246, Thr88, Asn 81 , His250, Met177, Asn253, Met 270, Glu169 and Phe168, according to the numbering of the adenosine A2A receptor as set out in Figure 13. Table (i)Of particular interest are Ser277 and His278 which bind uniquely to agonists and are thought to play a key role in the activation of the receptor.

In another example, the selected coordinates may comprise atoms of one or more amino acid residues that contribute to the NECA binding site of the A2A receptor. For example, amino acid residues contributing to the NECA binding site include amino acid residues Ile66, Ala63, He 274, Val84, His278, Ser277, Leu249, Trp246, Thr88, Asn181 , His250, Met177, Asn253, Met 270, Glu169, Phe168, Leu85 and Tyr9, according to the numbering of the adenosine A2A receptor sequence as set out in Figure 13. Thus the selected coordinates may comprise one or more atoms from any one or more (e.g. at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15, 16, 17 or 18 amino acid residues) of amino acid residues Ile66, Ala63, lie 274, Val84, His278, Ser277, Leu249, Trp246, Thr88, Asn181, His250, Met177, Asn253, Met 270, Glu169, Phe168, Leu85 or Tyr9, according to the numbering of the adenosine A2A receptor as set out in Figure 13. Table (i)Of particular interest are Ser277 and His278 which bind uniquely to agonists and are thought to play a key role in the activation of the receptor.

In a further example, the selected coordinates may comprise atoms of one or more amino acid residues of the ligand binding pocket where agonist ribose groups make polar interactions with concerved residues in H7 and non-polar residues in H3 (i.e., Val84 and Leu85 for the A2A-GL31 NECA-bound structure; Val84 and Thr88 for the adenosine-bound A2A-GL.3I structure. The conserved residues in H7 interacting with agonist ribose group(s) are: amino acid residues Ser277 and His278, according to the numbering of the adenosine A2A receptor sequence as set out in Figure 13. Thus, the selected coordinates may comprise one or more atoms from any one or more of Ser277 and/or His278 according to the numbering of the adenosine A2A receptor as set out in Figure 13 (e.g. at least one or both of said amino acid residues).

In global alignments of the A2A -GL31 structures with A2A-T4L (A^ with T4 lysozyme inserted into inner loop 3) bound to the inverse agonist ZM241385 based on those residues in the region of the ligand binding pocket that show the closest structural homology (Fig. 1) the most significant differences between the two structures are seen in a distortion and a 2 A shift primarily along the helical axis of H3, a bulge in H5 (resulting from non-helical backbone conformation angles of residues Cys185 and Val186) that shifts residues into the binding pocket by up to 2 A and also a change in conformation of the cytoplasmic ends of H5, H6 and H7 (Fig. 1). Hence, in another example, the selected coordinates may comprise atoms of one or more amino acids which define: the helical axis of H3; a bulge in H5 resulting from non-helical backbone conformation angles of residues Cys185 and Val186; cytoplasmic ends of H5; a change in conformation of the cytoplasmic ends of H6; and/or a change in conformation of the cytoplasmic ends of H7.

In the simplest model for the conformational dynamics of GPCRs¹⁰ there is an equilibrium between two states, R and R*. The inactive state R preferentially binds inverse agonists and the activated state R* preferentially binds agonists¹¹. By 'state R' or 'R state' we include GPCRs in their inactive state. By 'state R*' or 'R* state' we include GPCRs in their active state. However, GPCRs can also adopt numerous states of partial activation/inactivation intermediate between state 'R' and state 'R^*'. In particular, by intermediate state we refer to the partially activated state of adenosine/NECA-bound A2A-GL31 described in the Examples section, below.

Hence, In a further example, the selected coordinates may comprise atoms of one or more amino acids which define regions of the ligand receptor involved in the transition between R and R*. Mutations in these regions provide a thermostabilising effect to NECA-bound A2A- GL31. Thus, the selected coordinates may comprise atoms of one or more of amino acid resides Ala48 and/or Ala89, according to the numbering of the adenosine A∑A receptor as set out in Figure 13 or the selected coordinates may comprise atoms of one or more of amino acid resides Leu48 and/or Gln89, according to the numbering of the adenosine A_¾ receptor as set out in Figure 14. In a still further example, the selected coordinates may comprise atoms of one or more amino acids which define regions of the receptor-lipid interface that provide a thermostabilising effect to NECA-bound A2A-GL31. Thus, the selected coordinates may comprise atoms of one or more of amino acid resides Leu54 and/or Ala65, according to the numbering of the adenosine A2A receptor as set out in Figure 13 or the selected coordinates may comprise atoms of one or more of amino acid resides Ala54 and/or Thr65, according to the numbering of the adenosine A^ receptor as set out in Figure 14.

It is appreciated that the selected coordinates may comprise any atoms of particular interest including atoms mentioned in any one or more of the above examples, or as listed in the Examples section, below. It is appreciated that the selected coordinates may correspond to atoms from a particular structural region (e.g. helix and/or loop) of the adenosine A2A receptor. By the helices and loop regions of the adenosine A_M receptor we mean the following:

Helix 1 Residues 7-32

Helix 2 Residues 39-67

Helix 3 Residues 73-107

Helix 4 Residues 119-140

Helix 5 Residues 174-213

Helix 6 Residues 224-258

Helix 7 Residues 266-291

ICL1 Residues 33-38

ECL1 Residues 68-72

ICL2 Residues 108-118

ECL2 Residues 141-173

ICL3 Residues 214-213

ECL3 Residues 259-265

However, it will be appreciated that there are different criteria for which residues are considered to be in a helical conformation depending on phi and psi angles. Moreover, when comparing the adenosine A2A receptor to other structures, some residues may be missing in one or other of the structures and some residues may be considered helical in one structure but not the other. Further, the loop regions may be defined as amino acid structures that join alpha helices (as above) or may be defined as amino acid structures that are predicted to be outside of the membrane. Therefore the limits above are not to be construed as absolute, but rather may vary according to the criteria used. Nevertheless, for the purposes of the comparisons set out below, we have used the above-mentioned definitions of helices and loops. Preferably, the selected coordinates include at least 2% or 5% C-a atoms, and more preferably at least 10% C-a atoms. Alternatively or additionally, the selected coordinates include at least 10% and more preferably at least 20% or 30% backbone atoms selected from any combination of the nitrogen, C-a, carbonyl C and carbonyl oxygen atoms. It is appreciated that the coordinates of the adenosine A2A receptor used in the invention may be optionally varied and a subset of the coordinates or the varied coordinates may be selected (and constitute selected coordinates). Indeed, such variation may be necessary in various aspects of the invention, for example in the modelling of protein structures and in the fitting of various binding partners to the adenosine A_¾ receptor structure. Protein structure variability and similarity is routinely expressed and measured by the root mean square deviation (rmsd), which measures the difference in positioning in space between two sets of atoms. The rmsd measures distance between equivalent atoms after their optimal superposition. The rmsd 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 over C-a atoms only.

The least-squares algorithms used to calculate rmsd are well known in the art and include those described by Rossman and Argos {J Biol Chem, (1975) 250:7525), Kabsch (Acta Cryst (1976) A92:922; Acta Cryst (1978) A34:827-828), Hendrickson (Acta Cryst (1979) A35: 158), McLachan (J Mol Biol (1979) 128:49) and Kearsley (Acta Cryst (1989) A45:208). Both algorithms based on iteration in which one molecule is moved relative to the other, such as that described by Ferro and Hermans (Acta Cryst (1977) A33:345-347), and algorithms which locate the best fit directly (e.g. Kabsch's methods) may be used. Methods of comparing proteins structures are also discussed in Methods of Enzymology, vol 115: 397- 420.

Typically, rmsd values are calculated using coordinate fitting computer programs and any suitable computer program known in the art may be used, for example MNYFIT (part of a collection of programs called COMPOSER, Sutcliffe et al (1987) Protein Eng 1 :377-384). Other programs also include LSQMAN (Kleywegt & Jones (1994) A super position, CCP4/ESF-EACBM, Newsletter on Protein Crystallography, 31 : 9-14), LSQKAB (Collaborative Computational Project 4. The CCP4 Suite: Programs for Protein Crystallography, Acta Cryst (1994) D50:760-763), QUANTA (Jones et al, Acta Cryst (1991) A47:110-1 19 and commercially available from Accelrys, San Diego, CA), Insight (Commercially available from Accelrys, San Diego, CA), Sybyl® (commercially available from Tripos, Inc., St Louis) and O (Jones et al., Acta Cryst (1991) A47: 10-1 19).

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 as is well known in the art. The atomic coordinates can then be superimposed according to this alignment and an rmsd value calculated. The program Sequoia (Bruns et al (1999) J Mol Biol 288(3):427-439) performs the alignment of homologous protein sequences, and the superposition of homologous protein atomic coordinates. Once aligned, the rmsd can be calculated using programs detailed above. When the sequences are identical or highly similar, 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. RMSD analysis of all atoms and of residue backbone atoms (i.e. the nitrogen-carbon-carbon backbone atoms of the protein) between the adenosine A2A-GL31 receptor and the other structures (for example, the Jaakola et al A_¾ structure⁸ (PDB code: 3EML)) was performed using a Maestro script. Global alignments of the A2AR-GL31 structures with A2A bound to the inverse agonist ZM241385⁴⁵ were performed based on those residues in the region of the ligand binding pocket that show the closest structural homology (Fig. 1 and Example 1). This gave an rmsd in Calpha positions of 0.66 A for the 96 atoms selected (residues 16-21 in H1 , 51-70 in H2 and ECL1 , 132-140 in H4 and ECL2, 142-146 in ECL2, 166-182 in ECL2 and H5 and 245- 283 in H6, ECL3 and H7), which include all residues involved in binding either adenosine or NECA, with the exception of those in H3. Thus in one embodiment, the coordinates or selected coordinates of Table (i) may be optionally varied within an rmsd of the structurally homologous region of the ligand binding pocket (i.e. residues 16-21 in H1 , 51-70 in H2 and ECL1 , 132-140 in H4 and ECL2, 142-146 in ECL2, 166-182 in ECL2 and H5 and 245-283 in H6, ECL3 and H7) of not more than 0.66 A. Preferably, the coordinates or selected coordinates are varied within an rmsd of residue backbone atoms of not more than 0.65 A, 0.64 A, 0.63 A, 0.62 A or 0.61 A and more preferably not more than 0.6 A, 0.55 A, 0.5 A, 0.4 A, 0.3 A, 0.2 A or 0.1 A. Alignments of the A2AR-GL31 structures with: (A) A₂A bound to the inverse agonist ZM241385⁴⁵, (B) the Jaakola et al AZA structure⁸ (PDB code: 3EML), and (C) the Xu structure (PDB code: 3QAK) were also performed based on those residues common to all structures (7-148, 158-208, 223-305). For the Jaakola et al A_2A structure⁸ (PDB code: 3QAK) this gave an rmsd of residue backbone atoms of 1.88. For the Jaakola et al A_2A structure⁸ (PDB code: 3QAK) this gave an rmsd for all atoms of 2.34. For structure A_¾ bound to the inverse agonist ZM241385 this gave an rmsd of residue backbone atoms of 1.85. For structure A_¾ bound to the inverse agonist ZM241385⁴⁵ this gave an rmsd for all atoms of 2.13. For the Xu structure (PDB code: 3QAK) this gave an rmsd for residue backbone atoms of 1.08 A . Thus, in one embodiment, the coordinates or selected coordinates of Table (i) may be optionally varied within an rmsd of residue backbone atoms of not more than 1.08 A. Preferably, the coordinates or selected coordinates are varied within an rmsd of residue backbone atoms of not more 1.0 A, and more preferably not more than 0.9 A, 0.8 A, 0.7 A, 0.6 A, 0.5 A, 0.4 A, 0.3 A, 0.2 A or 0.1 A.

For the Xu structure (PDB code: 3QAK) this gave an rmsd for all atoms of 1.58. Thus, in one embodiment, the coordinates or selected coordinates of Table (i) may be optionally varied within an rmsd of all atoms of not more than 1.58 A. Preferably, the coordinates or selected coordinates are varied within an rmsd of residue atoms of not more than 1.5A or 1.4 A and more preferably not more than 1.3 A, 1.2 A, 1.1 A or 1.0 A, and even more preferably not more than 0.9 A, 0.8 A, 0.7 A, 0.6 A, 0.5 A, 0.4 A, 0.3 A, 0.2 A or 0.1 A.

Alignments of the A2A -GL31 structures with: (A) A_¾ bound to the inverse agonist ZM241385⁴⁵, (B) the Jaakola et al structure⁸ (PDB code: 3EML), and (C) the Xu structure (PDB code: 3QAK) were also performed based on those residues of the adenosine agonist complex (i.e., ligand-bound A2A -GL31) in the agonist binding pocket with exception of those in TM3: (residues 16-21 in H1 , 51-70 in H2 and ECL1 , 132-140 in H4 and ECL2, 142-146 in ECL2, 166-182 in ECL2 and H5 and 245-283 in H6, ECL3 and H7).

For the Jaakola et al A¾ structure⁸ (PDB code: 3EML) this gave an rmsd of residue backbone atoms of 1.89. For the Jaakola et al A_2A structure⁸ (PDB code: 3EML) this gave an rmsd for all atoms of 2.39. For structure A_¾ bound to the inverse agonist ZM241385⁴⁵ this gave an rmsd of residue backbone atoms of 1.84. For structure A^ bound to the inverse agonist ZM241385⁴⁵ this gave an rmsd for all atoms of 2.30.

For the Xu structure (PDB code: 3QAK) this gave an rmsd of residue backbone atoms of 1.08 A. Thus, in one embodiment, the coordinates or selected coordinates of Table (i) may be optionally varied within an rmsd of the residue backbone atoms of the adenosine agonist complex of not more than 1.08 A. Preferably, the coordinates or selected coordinates are varied within an rmsd of residue backbone atoms of not more 1.0 A, and more preferably not more than 0.9 A, 0.8 A, 0.7 A, 0.6 A, 0.5 A, 0.4 A, 0.3 A, 0.2 A or 0.1 A.

For the Xu structure (PDB code: 3QAK) this gave an rmsd for all atoms of 1.59. Thus, in one embodiment, the coordinates or selected coordinates of Table (i) may be optionally varied within an rmsd of all atoms of the adenosine agonist complex of not more than 1.59 A. Preferably, the coordinates or selected coordinates are varied within an rmsd of all atoms of not more than 1.5A or 1.4 A and more preferably not more than 1.3 A, 1.2 A, 1.1 A or 1.0 A, and even more preferably not more than 0.9 A, 0.8 A, 0.7 A, 0.6 A, 0.5 A, 0.4 A, 0.3 A, 0.2 A or 0.1 A.

Alignments of the A2A -GL31 structures with: (A) A_∑A bound to the inverse agonist ZM241385⁴⁵, (B) the Jaakola et a/ A^ structure⁸ (PDB code: 3EML), and (C) the Xu structure (PDB code: 3QAK) were also performed based on those residues of the adenosine binding pocket (i.e., residues Ile66, Ala63, lie 274, Val84, His278, Ser277, Leu249, Trp246, Thr88, Asn 81 , His250, Met177, Asn253, Met 270, Glu 69 and Phe168).

For the Jaakola et al A¾ structure⁸ (PDB code: 3EML) this gave an rmsd of side chain atoms of 1.37.

For structure A₂A bound to the inverse agonist ZM241385⁴⁵ this gave an rmsd ifor side chain atoms of 1.84.

For the Xu structure (PDB code: 3QAK) this gave an rmsd for side chain atoms of 1.36. Thus, in one embodiment, the coordinates or selected coordinates of Table (i) may be optionally varied within an rmsd for side chain atoms of the adenosine binding pocket (i.e., residues Ile66, Ala63, lie 274, Val84, His278, Ser277, Leu249, Trp246, Thr88, Asn181 , His250, Met177, Asn253, Met 270, Glu169 and Phe168) of not more than 1.36 A. Preferably, the coordinates or selected coordinates are varied within an rmsd of residue side chain atoms of not more 1.0 A, and more preferably not more than 1.3 A , 1.2 A , 1.1 A , 1.0 A , 0.9 A, 0.8 A, 0.7 A, 0.6 A, 0.5 A, 0.4 A, 0.3 A, 0.2 A or 0.1 A.

Alignments of the A2AR-GL31 structures with: (A) A2A-T4L (A₂AR with T4 lysozyme inserted into inner loop 3) bound to the inverse agonist ZM241385, (B) the Jaakola er al A2A structure⁸ (PDB code: 3EML), and (C) the Xu structure (PDB code: 3QAK) were also performed based on those residues of the NECA binding pocket (i.e., residues Ile66, Ala63, lie 274, Val84, His278, Ser277, Leu249, Trp246, Thr88, Asn181 , His250, Met177, Asn253, Met 270, Glu169, Phe168, Leu85 and Tyr9).

For the Jaakola et al A2A structure⁸ (PDB code: 3EML) this gave an rmsd for side chain atoms of 1.38.

For structure A_¾ bound to the inverse agonist ZM241385⁴⁵ this gave an rmsdfor side chain atoms of 2.8. For the Xu structure (PDB code: 3QAK) this gave an rmsd for side chain atoms of 1.27. Thus, in one embodiment, the coordinates or selected coordinates of Table (i) may be optionally varied within an rmsd for side chain atoms of the NECA binding pocket (i.e., residues Ile66, Ala63, lie 274, Val84, His278, Ser277, Leu249, Trp246, Thr88, Asn181 , His250, Met177, Asn253, Met 270, Glu169, Phe168, Leu85 and Tyr9) of not more than 1.27 A. Preferably, the coordinates or selected coordinates are varied within an rmsd of residue side chain atoms of not more 1.0 A, and more preferably not more than 1.2 A , 1.1 A , 1.0 A , 0.9 A, 0.8 A, 0.7 A, 0.6 A, 0.5 A, 0.4 A, 0.3 A, 0.2 A or 0.1 A.

In this aspect of the invention, the coordinates of the adenosine A2A receptor structure are used to predict a three dimensional representation of a target protein of unknown structure, or part thereof, by modelling. By "modelling", we mean the prediction of structures using computer-assisted or other de novo prediction of structure, based upon manipulation of the coordinate data from Table (i) or selected coordinates thereof. The target protein may be any protein that shares sufficient sequence identity to the human adenosine A_¾ receptor such that its structure can be modelled by using the adenosine A_¾ receptor coordinates of Table (i). It will be appreciated that if a structural representation of only a part of the target protein is being modelled, for example a particular domain, the target protein only has to share sufficient sequence identity to the adenosine A_¾ receptor over that part.

It has been shown for soluble protein domains that their three dimensional structure is broadly conserved above 20% amino acid sequence identity and well conserved above 30% identity, with the level of structural conservation increasing as amino acid sequence identity increases up to 100% (Ginalski, K. Curr Op Struc Biol (2006) 16, 172-177). Thus, it is preferred if the target protein, or part thereof, shares at least 20% amino acid sequence identity with the human adenosine A_¾ receptor sequence provided in Figure 16, and more preferably at least 30%, 40%, 50%, 60%, 70%, 80% or 90% sequence identity, and yet more preferably at least 95% or 99% sequence identity. It will be appreciated therefore that the target protein may be an adenosine A_¾ receptor analogue or homologue.

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 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. By a human adenosine A a, receptor homologue, we include a protein with at least 20%, 25%, 30%, 35%, 40%, 45% or at least 50% amino acid sequence identity with the sequence of adenosine A_¾ receptor provided in Figure 16, preferably at least 55%, 60%, 65%, 70%, 75% or 80% amino acid sequence identity and more preferably 85%, 90%, 95% or 99% amino acid sequence identity. This includes polymorphic forms of adenosine A∑A receptors, e.g. mutants and adenosine A_¾ receptors from other species as well as other adenosine receptors such as A^ A¾, A₂B, A₃. Thus an adenosine A2A receptor homologue would include a human adenosine A_{1 ;} A₂B or A₃ receptor.

Sequence identity may be measured by the use of algorithms such as BLAST or PSI-BLAST (Altschul et al, NAR (1997), 25, 3389-3402) or methods based on Hidden Markov Models (Eddy S et al, J Comput Biol (1995) Spring 2 (1) 9-23). Typically, the percent sequence identity between two polypeptides may be determined using any suitable computer program, for example the GAP program of the University of Wisconsin Genetic Computing Group and it will be appreciated that percent identity is calculated in relation to polypeptides whose sequence has been aligned optimally. The alignment may alternatively be carried out using the Clustal W program (Thompson et al., 1994). The parameters used may be as follows: Fast pairwise alignment parameters: K-tuple(word) size; 1 , window size; 5, gap penalty; 3, number of top diagonals; 5. Scoring method: x percent. Multiple alignment parameters: gap open penalty; 10, gap extension penalty; 0.05. Scoring matrix: BLOSUM. In one embodiment the target protein is an integral membrane protein. By "integral membrane protein" we mean a protein that is permanently integrated into the membrane and can only be removed using detergents, non-polar solvents or denaturing agents that physically disrupt the lipid bilayer. Examples include receptors such as GPCRs, the T-cell receptor complex and growth factor receptors; transmembrane ion channels such as ligand- gated and voltage gated channels; transmembrane transporters such as neurotransmitter transporters; enzymes; carrier proteins; and ion pumps.

The amino acid sequences (and the nucleotide sequences of the cDNAs which encode them) of many membrane proteins are readily available, for example by reference to GenBank. For example, Foord et al supra gives the human gene symbols and human, mouse and rat gene IDs from Entrez Gene (http://www.ncbi.nlm.nih.gov/entrez) for GPCRs. It should be noted, also, that because the sequence of the human genome is substantially complete, the amino acid sequences of human membrane proteins can be deduced therefrom.

In a preferred embodiment, the target protein is a GPCR.

Suitable GPCRs include, but are not limited to adenosine receptors, β-adrenergic receptors, purinergic receptors, dopaminergic receptors, chemokine receptors and muscarinic receptors. Other suitable GPCRs are well known in the art and include those listed in Hopkins & Groom supra. In addition, the International Union of Pharmacology produce a list of GPCRs (Foord et al (2005) Pharmacol. Rev. 57, 279-288, incorporated herein by reference and this list is periodically updated at http://www.iuphar- db.org/GPCR/ReceptorFamiliesForward). It will be noted that GPCRs are divided into different classes, principally based on their amino acid sequence similarities. They are also divided into families by reference to the natural ligands to which they bind. All GPCRs are included in the scope of the invention and their structure may be modelled by using the coordinates of the adenosine A2A receptor.

Although the target protein may be derived from any source, it is particularly preferred if it is from a eukaryotic source. It is particularly preferred if it is derived from a vertebrate source such as a mammal. It is particularly preferred if the target protein is derived from rat, mouse, rabbit or dog or non-human primate or man. Typically, modelling a structural representation of a target is done by homology modelling whereby homologous regions between the adenosine A2A receptor and the target protein are matched and the coordinate data of the adenosine A2A receptor used to predict a structural representation of the target protein.

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.

Typically, the method involves comparing the amino acid sequences of adenosine A2A receptor with a target 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.

Homology between amino acid sequences can be determined using commercially available algorithms known in the art. For example, the programs BLAST, gapped BLAST, BLASTN, PSI-BLAST, BLAST 2 and WU- BLAST (provided by the National Center for Biotechnology Information) can be used to align homologous regions of two, or more, amino acid sequences. These may be used with default parameters to determine the degree of homology between the amino acid sequence of the adenosine A_¾ receptor and other target proteins which are to be modelled.

Preferred for use according to the present invention is the WU-BLAST (Washington University BLAST) version 2.0 software. WU-BLAST version 2.0 executable programs for several UNIX platforms can be downloaded from ftp ://blast. wustl. edu/blast/executables. This program is based on WU-BLAST version 1.4, which in turn is based on the public domain NCBI-BLAST version 1.4 (Altschul and Gish, 1996, Local alignment statistics, Doolittle ed., Methods in Enzymology 266: 460-480; Altschul et al., 1990, Basic local alignment search tool, Journal of Molecular Biology 215: 403-410; Gish and States, 1993, Identification of protein coding regions by database similarity search, Nature Genetics 3: 266- 272; Karlin and Altschul, 1993, Applications and statistics for multiple high-scoring segments in molecular sequences, Proc. Natl. Acad. Sci. USA 90: 5873-5877; all of which are incorporated by reference herein). In all search programs in the suite the gapped alignment routines are integral to the database search itself. Gapping can be turned off if desired. The default penalty (Q) for a gap of length one is Q=9 for proteins and BLASTP, and Q=10 for BLASTN, but may be changed to any integer. The default per-residue penalty for extending a gap (R) is R=2 for proteins and BLASTP, and R=10 for BLASTN, but may be changed to any integer. Any combination of values for Q and R can be used in order to align sequences so as to maximize overlap and identity while minimizing sequence gaps. The default amino acid comparison matrix is BLOSUM62, but other amino acid comparison matrices such as PAM can be utilized.

Once the amino acid sequences of adenosine A_¾ receptor and the target protein of unknown structure have been aligned, the structures of the conserved amino acids in the structural representation of the adenosine A_¾ receptor may be transferred to the corresponding amino acids of the target protein. For example, a tyrosine in the amino acid sequence of adenosine A2A receptor may be replaced by a phenylalanine, the corresponding homologous amino acid in the amino acid sequence of the target protein.

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. Typically, the predicted three dimensional structural representation will be one in which favourable interactions are formed within the target protein and/or so that a low energy conformation is formed ("High resolution structure prediction and the crystallographic phase problem" Qian et al (2007) Nature 450; 259-264; "State of the art in studying protein folding and protein structure production using molecular dynamics methods" Lee et al (2001) J of Mol Graph & Modelling 19(1): 146-149).

Whereas it is preferred to base homology modelling on homologous amino acid sequences, it is appreciated that some proteins have low sequence identity (e.g. family B and C GPCRs) and at the same time are very similar in structure. Therefore, where at least part of the structure of the target protein is known, homologous regions can also be identified by comparing structures directly. Homology modelling as such is a technique well known in the art (see e.g. Greer, (Science, Vol. 228, (1985), 1055), and Blundell et al (Eur. J. Biochem, Vol. 172, (1988), 513)). The techniques described in these references, as well as other homology modelling techniques generally available in the art, may be used in performing the present invention.

Typically, homology modelling is performed using computer programs, for example SWISS- MODEL available through the Swiss Institute for Bioinformatics in Geneva, Switzerland; WHATIF available on EMBL servers; Schnare et al. (1996) J. Mol. Biol, 256: 701-719; Blundell et al. (1987) Nature 326: 347-352; Fetrow and Bryant (1993) Bio/Technology 11 :479-484; Greer (1991 ) Methods in Enzymology 202: 239-252; and Johnson et al (1994) Crit. Rev. Biochem. Mol Biol. 29:1-68. An example of homology modelling is described in Szklarz G. D (1997) Life Sci. 61 : 2507-2520.

Thus, in an embodiment of the first aspect of the invention, the method further comprises aligning the amino acid sequence of the target protein of unknown structure with the amino acid sequence of adenosine A_¾ receptor listed in Figure 13 or 14 to match homologous regions of the amino acid sequences, and subsequently modelling the structural representation of the target protein by modelling the structural representation of the matched homologous regions of the target protein on the corresponding regions of the adenosine A_¾ receptor to obtain a three dimensional structural representation for the target protein that substantially preserves the structural representation of the matched homologous regions.

The invention therefore provides a method of predicting a three dimensional structural representation of a target protein of unknown structure, or part thereof, comprising:

providing the coordinates of the human adenosine A_¾ receptor structure listed in Table (i), optionally varied by a root mean square deviation of residue backbone atoms of not more than .08 A, or selected coordinates thereof; and

predicting the three-dimensional structural representation of the target protein, or part thereof, by modelling the structural representation on all or the selected coordinates of the adenosine A_∑A receptor;

aligning the amino acid sequence of a target protein of unknown structure or part thereof with the amino acid sequence of adenosine Aa_, receptor listed in Figure 13 or 14 or part thereof to match homologous regions of the amino acid sequences;

modelling the structure of the matched homologous regions of the target protein on the corresponding regions of the adenosine A_¾ receptor structure as defined by Table (i), optionally varied by a root mean square deviation of residue backbone atoms of not more than 1.08 A, or selected coordinates thereof; and predicting a three dimensional structural representation for the target protein which substantially preserves the structure of the matched homologous regions.

The coordinate data of Table (i), or selected coordinates thereof, will be particularly advantageous for homology modelling of other GPCRs. For example, since the protein sequence of adenosine A2A receptor and another GPCR can be aligned relative to each other, it is possible to predict structural representations of the structures of other GPCRs, particularly in the regions of the transmembrane helices and ligand binding region, using the adenosine A2A receptor coordinates.

The coordinate data of the adenosine A_¾ receptor can also be used to predict the crystal structure of target proteins where X-ray diffraction data or NMR spectroscopic data of the protein has been generated and requires interpretation in order to provide a structure. A second aspect of the invention provides a method of predicting the three dimensional structural representation of a target protein of unknown structure, or part thereof, comprising: providing the coordinates of the adenosine A_¾ receptor structure listed in Table (i), optionally varied by a root mean square deviation of residue backbone atoms of not more than 1.08 A, or selected coordinates thereof; and either (a) positioning the coordinates in the crystal unit cell of the protein so as to predict its structural representation, or (b) assigning NMR spectra peaks of the protein by manipulating the coordinates.

Thus, where X-ray crystallographic or NMR spectroscopic data is provided for a target protein of unknown structure, the coordinate data of Table (i) may be used to interpret that data to predict a likely structure using techniques well known in the art including phasing, in the case of X-ray crystallography, and assisting peak assignments in the case of NMR spectra.

A three dimensional structural representation of any part of any target protein that is sufficiently similar to any portion of the adenosine A2A receptor can be predicted by this method. Typically, the target protein or part thereof has at least 20% amino acid sequence identity with any portion of adenosine A_¾ receptor, such as at least 30% amino acid sequence identity or at least 40% or 50% or 60% or 70% or 80% or 90% sequence identity. For example, the coordinates may be used to predict the three-dimensional representations of other crystal forms of adenosine A2A receptor, other adenosine A2A receptors, adenosine A2A receptor mutants or co-complexes of an adenosine Az¾ receptor. Other suitable target proteins are as defined with respect to the first aspect of the invention.

One method that may be employed for these purposes is molecular replacement which is well known in the art and described, for example, in Evans & McCoy {Acta Cryst, 2008, D64:1-10), McCoy (Acta Cryst, 2007, D63:32-42) and McCoy et al (J of App Cryst, 2007, 40:658-674). Molecular replacement enables the solution of the crystallographic phase problem by providing initial estimates of the phases of the new structure from a previously known structure, as opposed to the other major methods for solving the phase problem, i.e. experimental methods (which measure the phase from isomorphous or anomalous differences) or direct methods (which use mathematical relationships between reflection triplets and quartets to bootstrap a phase set for all reflections from phases for a small or random 'seed' set of reflections.) Compared to molecular replacement, such methods are time consuming and generally hinder the solution of crystal structures. Thus molecular replacement provides an accurate structural form for an unknown crystal more quickly and efficiently than attempting to determine such information ab initio.

Accordingly, the invention involves generating a preliminary model of a target protein whose structure coordinates are unknown, by orienting and positioning the relevant portion of the adenosine A_¾ receptor according to Table (i) within the unit cell of a crystal of the target protein so as best to account for the observed X-ray diffraction pattern of the crystal of the target protein. Phases can be calculated from this model and combined with the observed X- ray diffraction pattern amplitudes to generate an electron density map of the target protein's structure. This, in turn, can be subjected to any well-known model building and structure refinement techniques to provide a final, accurate structural representation of the target protein (E. Lattman, "Use of the Rotation and Translation Functions", in Meth. Enzymol., 115, pp. 55-77 (1985); M. G. Rossmann, ed., "The Molecular Replacement Method", Int. Sci. Rev. Ser., No. 13, Gordon & Breach, New York (1972)). Thus the invention includes a method of predicting a three dimensional structural representation of a target protein of unknown structure, or part thereof, comprising: providing the coordinates of the adenosine A2A receptor structure, optionally varied by a root mean square deviation of residue backbone atoms of not more than 1.08 A, or selected coordinates thereof; providing an X-ray diffraction pattern of the target protein; and using the coordinates to predict at least part of the structure coordinates of the target protein. In an embodiment, the X-ray diffraction pattern of the target protein is provided by crystallising the target protein unknown structure; and generating an X-ray diffraction pattern from the crystallised target protein. Thus, the invention also provides a method of method of predicting a three dimensional structural representation of a target protein of unknown structure comprising the steps of (a) crystallising the target protein; (b) generating an X-ray diffraction pattern from the crystallised target protein; (c) applying the coordinates of the adenosine A_¾ receptor structure, optionally varied by a root mean square deviation of residue backbone atoms of not more than 1.08 A, or selected coordinates thereof, to the X- ray diffraction pattern to generate a three-dimensional electron density map of the target protein, or part thereof; and (d) predicting a three dimensional structural representation of the target protein from the three-dimensional electron density map.

Examples of computer programs known in the art for performing molecular replacement include CNX (Brunger AT.; 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, CA), MOLREP (A.Vagin, A.Teplyakov, MOLREP: an automated program for molecular replacement, J Appl Cryst (1997) 30, 1022-1025, part of the CCP4 suite), AMoRe (Navaza, J. (1994). AMoRe: an automated package for molecular replacement. Acta Cryst A50, 157- 163), or PHASER (part of the CCP4 suite).

Preferred selected coordinates of the adenosine A_¾ receptor are as defined above with respect to the first aspect of the invention.

The invention may also be used to assign peaks of NMR spectra of target proteins, by manipulation of the data of Table (i) (J Magn Reson (2002) 157(1 ): 1 19-23).

The coordinates of the adenosine A2A receptor of Table (i), optionally varied by a root mean square deviation of residue backbone atoms of not more than 1.08 A, or selected coordinates thereof may be used in the provision, design, modification or analysis of binding partners of adenosine A2A receptors. Such a use will be important in drug design.

By adenosine A₂A receptor we mean any adenosine A2A receptor which has at least 75% sequence identity with human adenosine A2A receptor as well as adenosine A2A receptors from other species and mutants thereof. Preferably, the adenosine A2A receptor has at least 80% amino acid sequence identity to human adenosine A2A receptor, and more preferably at least 85%, 90%, 95% or 99% amino acid sequence identity. By "binding partner" we mean any molecule that binds to an adenosine A2A receptor. Preferably, the molecule binds selectively to the adenosine A_M receptor. For example, it is preferred if the binding partner has a Kd value (dissociation constant) which is at least five or ten times lower (i.e. higher affinity) than for at least one other adenosine receptor (A_{1 f} A_2B or A₃), and preferably more than 100 or 500 times lower. More preferably, the binding partner of an adenosine A2A receptor has a d value more than 000 or 5000 times lower than for at least one other adenosine receptor. However, it will be appreciated that the limits will vary dependent upon the nature of the binding partner. Thus, typically, for small molecule binding partners, the binding partner typically has a K_d value which is at least 10 times or 50 times or 100 times lower than for at least one other adenosine receptor. Typically, for antibody binding partners, the binding partner typically has a Kd value which is at least 500 or 1000 times lower than for at least one other adenosine receptor. Kd values can be determined readily using methods well known in the art and as described, for example, below.

At equilibrium Kd=[R][L]/[RL]

where the terms in brackets represent the concentration of

· Receptor-ligand complexes [RL],

• unbound receptor [R], and

• unbound ("free") ligand [L].

In order to determine the Kd the value of these terms must be known. Since the concentration of receptor is not usually known then the Hill-Langmuir equation is used where Fractional occupancy= [L]/[L] + K_d.

In order to experimentally determine a K_d then, the concentration of free ligand and bound ligand at equilibrium must be known. Typically, this can be done by using a radio-labelled or fluorescently labelled ligand which is incubated with the receptor (present in whole cells or homogenised membranes) until equilibrium is reached. The amount of free ligand vs bound ligand must then be determined by separating the signal from bound vs free ligand. In the case of a radioligand this can be done by centrifugation or filtration to separate bound ligand present on whole cells or membranes from free ligand in solution. Alternatively a scintillation proximity assay is used. In this assay the receptor (in membranes) is bound to a bead containing scintillant and a signal is only detected by the proximity of the radioligand bound to the receptor immobilised on the bead.

It will be appreciated that the coordinates of the invention will also be useful in the analysis of solvent and ion interactions with an adenosine A2A receptor, which are important factors in drug design.

It is particularly preferred if the binding partner is a small molecule with a molecule weight of 1000 daltons or less, for example, less than 900 daltons, less than 800 daltons, less than 700 daltons, less than 600 daltons, less than 500 daltons, less than 450 daltons, less than 400 daltons, less than 350 daltons, less than 300 daltons, less than 250 daltons, less than 200 daltons, less than 150 daltons, less than 100 daltons, less than 50 daltons or less than 10 daltons. It is further preferred if the binding partner causes a change (i.e a modulation) in the level of biological activity of the adenosine A¾ receptor, i.e. it has functional agonist or antagonist activity, and therefore may have the potential to be a candidate drug. Thus, the binding partner may be any of a full agonist, a partial agonist, an inverse agonist or an antagonist of adenosine A2A receptor. The binding partner may bind to the orthosteric site, e.g. as defined by the adenosine, NECA or ZM241385 binding sites, or it may bind to an allosteric binding site. It is also appreciated that the binding partner may be one that modulates the ability of the adenosine A₂A receptor to dimerise. For example, the binding partner may bind to the dimerisation interface or bind to another region of the adenosine A2A receptor which nevertheless modulates dimerisation.

Accordingly, a third aspect of the invention provides a method for selecting or designing one or more binding partners of adenosine A_¾ receptor comprising using molecular modelling means to select or design one or more binding partners of the adenosine A2A receptor, wherein the three-dimensional structural representation of at least part of the human adenosine A2A receptor, as defined by the coordinates of adenosine A2A receptor of Table (i), optionally varied by a root mean square deviation of residue backbone atoms of not more than 1.08 A, or selected coordinates thereof, is compared with a three-dimensional structural representation of one or more candidate binding partners, and one or more binding partners that are predicted to interact with adenosine A2A receptor are selected. In order to provide a three-dimensional structural representation of a candidate binding partner, the binding partner structural representation may be modelled 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 structural representation of the binding partner.

The design of binding partners that bind to an adenosine A2A receptor generally involves consideration of two factors. First, the binding partner must be capable of physically and structurally associating with parts or all of an adenosine A¾ receptor binding region (e.g. ligand binding site or an allosteric binding site or dimerisation interface). Non-covalent molecular interactions important in this association include hydrogen bonding, van der Waals interactions, hydrophobic interactions and electrostatic interactions.

Second, the binding partner must be able to assume a conformation that allows it to associate with an adenosine A_¾ receptor binding region directly. Although certain portions of the binding partner will not directly participate in these associations, those portions of the binding partner may still influence the overall conformation of the molecule. This, in turn, may have a significant impact on potency. Such conformational requirements include the overall three-dimensional structure and orientation of the binding partner in relation to all or a portion of the binding region, or the spacing between functional groups of a binding partner comprising several binding partners that directly interact with the adenosine A2A receptor. Thus it will be appreciated that selected coordinates which represent a binding region of the adenosine A_¾ receptor, e.g. atoms from amino acid residues contributing to the adenosine binding site include amino acid residues Ile66, Ala63, lie 274, Val84, His278, Ser277, Leu249, Trp246, Thr88, Asn181 , His250, Met177, Asn253, Met 270, Glu169 and Phe168, may be used, or atoms from amino acid residues contributing to the NECA binding site include amino acid residues Ile66, Ala63, lie 274, Val84, His278, Ser277, Leu249, Trp246, Thr88, Asn181, His250, Met177, Asn253, Met 270, Glu169, Phe168, Leu85 and Tyr9 may be used. Additional preferences for the selected coordinates are as defined above with respect to the first aspect of the invention. Designing of binding partners can generally be achieved in two ways, either by the step wise assembly of a binding partner or by the de novo synthesis of a binding partner. As is described in more detail below, binding partners can also be identified by virtual screening. With respect to the step-wise assembly of a binding partner, several methods may be used. Typically the process begins by visual inspection of, for example, any of the binding regions on a computer representation of the adenosine A_¾ receptor as defined by the coordinates in Table (i) optionally varied within a rmsd of residue backbone atoms of not more than 1.08 A, or selected coordinates thereof. Selected binding partners, or fragments or moieties thereof may then be positioned in a variety of orientations, or docked, within the binding region. Docking may be accomplished using software such as QUANTA and Sybyl (Tripos Associates, St. Louis, Mo.), followed by, or performed simultaneously with, energy minimization, rigid-body minimization (Gshwend, supra) and molecular dynamics with standard molecular mechanics force fields, such as CHARMM and AMBER.

Specialized computer programs may also assist in the process of selecting binding partners or fragments or moieties thereof, as are known in the art and as detailed in WO2008/068534 incorporated herein by reference. Once suitable binding partners or fragments have been selected, they may be assembled into a single compound or complex. Assembly may be preceded by visual inspection of the relationship of the fragments to each other on the three-dimensional image displayed on a computer screen in relation to the structure coordinates of the adenosine A_¾ receptor. This would be followed by manual model building using software such as QUANTA or Sybyl. Useful programs known in the art (see, for example WO2008/068534 incorporated herein by reference) may aid connecting the individual chemical entities or fragments.

Thus the invention includes a method of designing a binding partner of an adenosine A₂A receptor comprising the steps of: (a) providing a structural representation of an adenosine A2A receptor binding region as defined by the coordinates of the human adenosine A_¾ receptor of Table (i), optionally varied by a root mean square deviation of residue backbone atoms of not more than 1.08 A, or selected coordinates thereof (b) using computational means to dock a three dimensional structural representation of a first binding partner in part of the binding region; (c) docking at least a second binding partner in another part of the binding region; (d) quantifying the interaction energy between the first or second binding partner and part of the binding region; (e) repeating steps (b) to (d) with another first and second binding partner, selecting a first and a second binding partner based on the quantified interaction energy of all of said first and second binding partners; (f) optionally, visually inspecting the relationship of the first and second binding partner to each other in relation to the binding region; and (g) assembling the first and second binding partners into a one binding partner that interacts with the binding region by model building.

As an alternative to the step-wise assembly of binding partners, binding partners may be designed as a whole or "de novo" using either an empty binding region or optionally including some portion(s) of a known binding partner(s). There are many de novo ligand design methods including: 1. LUDI (H.-J. Bohm, "The Computer Program LUDI: A New Method for the De Novo Design of Enzyme Inhibitors", J. Comp. Aid. Molec. Design, 6, pp. 61-78 (1992)). LUDI is available from Molecular Simulations Incorporated, San Diego, Calif; 2. LEGEND (Y. Nishibata et al., Tetrahedron, 47, p. 8985 (1991)). LEGEND is available from Molecular Simulations Incorporated, San Diego, Calif; 3. LeapFrog (available from Tripos Associates, St. Louis, Mo.); and 4. SPROUT (V. Gillet et al., "SPROUT: A Program for Structure Generation)", J. Comput. Aided Mol. Design, 7, pp. 127-153 (1993)). SPROUT is available from the University of Leeds, UK.

Other molecular modelling techniques may also be employed in accordance with this invention (see, e.g., N. C. Cohen et al., "Molecular Modeling Software and Methods for

Medicinal Chemistry, J. Med. Chem., 33, pp. 883-894 (1990); see also, M. A. Navia and M.

A. Murcko, "The Use of Structural Information in Drug Design", Current Opinions in Structural

Biology, 2, pp. 202-210 (1992); L. M. Balbes et al., "A Perspective of Modern Methods in

Computer-Aided Drug Design", in Reviews in Computational Chemistry, Vol. 5, K. B. Lipkowitz and D. B. Boyd, Eds., VCH, New York, pp. 337-380 (1994); see also, W. C. Guida,

"Software For Structure-Based Drug Design", Curr. Opin. Struct. Biology, 4, pp. 777-781

(1994)).

In addition to the methods described above in relation to the design of binding partners, other computer-based methods are available to select for binding partners that interact with adenosine A_¾ receptor .

For example the invention involves the computational screening of small molecule databases for binding partners that can bind in whole, or in part, to the adenosine A2A receptor. In this screening, the quality of fit of such binding partners to a binding region of an adenosine A2A receptor site as defined by the coordinates of the human adenosine A_M receptor of Table (i), optionally varied by a root mean square deviation of residue backbone atoms of not more than 1.08 A, or selected coordinates thereof, may be judged either by shape complementarity or by estimated interaction energy (E. C. Meng et al., J. Comp. Chem., 13, pp. 505-524 (1992)).

For example, selection may involve using a computer for selecting an orientation of a binding partner with a favourable shape complementarity in a binding region comprising the steps of: (a) providing the coordinates of adenosine A2A receptor of Table (i), optionally varied by a root mean square deviation of residue backbone atoms of not more than 1.08 A, or selected coordinates thereof and a three-dimensional structural representation of one or more candidate binding partners; (b) employing computational means to dock a first binding partner in the binding region; (c) quantitating the contact score of the binding partner in different orientations; and (d) selecting an orientation with the highest contact score. The docking may be facilitated by the contact score. The method may further comprise the step of generating a three-dimensional structural representation of the binding region and binding partner bound therein prior to step (b).

The method may further comprise the steps of: (e) repeating steps (b) through (d) with a second binding partner; and (f) selecting at least one of the first or second binding partner that has a higher contact score based on the quantitated contact score of the first or second binding partner.

In another embodiment, selection may involve using a computer for selecting an orientation of a binding partner that interacts favourably with a binding region comprising; a) providing the coordinates of the human adenosine A¾ receptor of Table (i), optionally varied by a root mean square deviation of residue backbone atoms of not more than 1.08 A, or selected coordinates thereof; b) employing computational means to dock a first binding partner in the binding region; c) quantitating the interaction energy between the binding partner and all or part of a binding region for different orientations of the binding partner; and d) selecting the orientation of the binding partner with the most favorable interaction energy.

The docking may be facilitated by the quantitated interaction energy and energy minimization with or without molecular dynamics simulations may be performed simultaneously with or following step (b). The method may further comprise the steps of: (e) repeating steps (b) through (d) with a second binding partner; and (f) selecting at least one of the first or second binding partner that interacts more favourably with a binding region based on the quantitated interaction energy of the first or second binding partner.

In another embodiment, selection may involve screening a binding partner to associate at a deformation energy of binding of less than -7 kcal/mol with an adenosine A2A receptor binding region comprising: (a) providing the coordinates of adenosine A2A receptor of Table (i), optionally varied by a root mean square deviation of residue backbone atoms of not more than 1.08 A, or selected coordinates thereof and employing computational means which utilise coordinates to dock the binding partner into a binding region; (b) quantifying the deformation energy of binding between the binding partner and the binding region; and (d) selecting a binding partner that associates with an adenosine A^ receptor binding region at a deformation energy of binding of less than -7 kcal/mol.

Determination of the three-dimensional structure of the adenosine A^ receptor provides important information about the binding sites of adenosine A2A receptors, particularly when comparisons are made with other adenosine receptors. This information may then be used for rational design and modification of adenosine A_¾ receptor binding partners, 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 using X-ray crystallographic analysis. These techniques are discussed in more detail below. Thus as a result of the determination of the adenosine A2A receptor three-dimensional structure, more purely computational techniques for rational drug design may also be used to design structures whose interaction with adenosine A2A receptor 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 adenosine A2A receptor structure in silico may be equally applied to both the human adenosine A₂A receptor structure of of Table (i), optionally varied by a root mean square deviation of residue backbone atoms of not more than 1.08 A, or selected coordinates thereof; and, by predicting the three- dimensional structural representation of a target protein, or part thereof, by modelling the structural representation on all or the selected coordinates of the adenosine A2A receptor or selected coordinates thereof, to the models of target proteins obtained by the first and second aspects of the invention. Thus having determined a conformation of a target protein, for example an adenosine A_¾ receptor, by the methods 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 adenosine A2A receptor will allow the generation of highly predictive pharmacophore models for virtual library screening or ligand design.

Accordingly, a fourth aspect of the invention provides a method for the analysis of the interaction of one or more binding partners with adenosine A_¾ receptor, comprising: providing a three dimensional structural representation of adenosine A^ receptor as defined by the coordinates of the human adenosine A2A receptor structure of Table (i), optionally varied by a root mean square deviation of residue backbone atoms of not more than 1.08 A, or selected coordinates thereof; providing a three dimensional structural representation of one or more binding partners to be fitted to the structural representation of adenosine A₂A receptor or selected coordinates thereof; and fitting the one of more binding partners to said structure.

This method of the invention is generally applicable for the analysis of known binding partners of adenosine A2A receptor, the development or discovery of binding partners of adenosine A2A receptor, the modification of binding partners of adenosine A2A receptor e.g. to improve or modify one or more of their properties, and the like. Moreover, the methods of the invention are useful in identifying binding partners that are selective for adenosine A2A receptors over other adenosine receptors. For example, comparing corresponding binding regions between adenosine A_¾ receptors and other adenosine receptors will facilitate the design of adenosine A∑A specific binding partners.

It will be desirable to model a sufficient number of atoms of the adenosine A_¾ receptor as defined by the coordinates of Table (i), optionally varied by a root mean square deviation of residue backbone atoms of not more than 1.08 A, or selected coordinates thereof, which represent a binding region, e.g. atoms from amino acid residues contributing to the adenosine binding site include amino acid residues Ile66, Ala63, He 274, Val84, His278, Ser277, Leu249, Trp246, Thr88, Asn181 , His250, Met177, Asn253, Met 270, Glu169 and Phe168, or atoms from amino acid residues contributing to the NECA binding site include amino acid residues Ile66, Ala63, He 274, Val84, His278, Ser277, Leu249, Trp246, Thr88, Asn181 , His250, Met177, Asn253, Met 270, Glu169, Phe168, Leu85 and Tyr9. Although every different binding partner bound by adenosine A_¾ receptor may interact with different parts of a binding region of the protein, the structure of the adenosine Aa_, receptor allows the identification of a number of particular sites which are likely to be involved in many of the interactions of adenosine A2A receptor with a drug candidate. Additional preferred selected coordinates are as described as above with respect to the first aspect of the invention.

In order to provide a three-dimensional structural representation of a binding partner to be fitted to the adenosine A_¾ receptor structure, the binding partner structural representation may be modelled 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 structural representation of the binding partner for fitting to the adenosine A¾ receptor structures of the invention.

By "fitting", is meant determining by automatic, or semi-automatic means, interactions between one or more atoms of a candidate binding partner and at least one atom of the adenosine A2A receptor structure of the invention, and calculating the extent to which such interactions are stable. Interactions include attraction and repulsion, brought about by charge, steric, lipophilic, considerations and the like. Charge and steric interactions of this type can be modelled computationally. An example of such computation would be via a force field such as Amber (Cornell et a/. A Second Generation Force Field for the Simulation of Proteins, Nucleic Acids, and Organic Molecules, Journal of the American Chemical Society, (1995), 117(19), 5179-97) which would assign partial charges to atoms on the protein and binding partner and evaluate the electrostatic interaction energy between a protein and binding partner atom using the Coulomb potential. The Amber force field would also assign van der Waals energy terms to assess the attractive and repulsive steric interactions between two atoms. Lipophilic interactions can be modeled using a variety of means. For example the ChemScore function (Eldridge M D; Murray C W; Auton T R; Paolini G V; Mee R P Empirical scoring functions: I. The development of a fast empirical scoring function to estimate the binding affinity of binding partners in receptor complexes, Journal of computer- aided molecular design (1997 Sep), 11 (5), 425-45) assigns protein and binding partner atoms as hydrophobic or polar, and a favourable energy term is specified for the interaction between two hydrophobic atoms. Other methods of assessing the hydrophobic contributions to ligand binding are available and these would be known to one skilled in the art. Other methods of assessing interactions are available and would be known to one skilled in the art of designing molecules. Various computer-based methods for fitting are described further herein.

More specifically, the interaction of a binding partner with the adenosine A_¾ receptor structure of the invention can be examined through the use of computer modelling 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, (1982) J.Mol.Biol., 161 , 269-288; Makino et al, (1997) J.Comput.Chem., 18, 1812-1825), AUTODOCK (Goodsell et al, (1990) Proteins, 8, 195-202, Morris et al, (1998) J.Comput.Chem., 19, 1639- 1662.), FlexX, (Rarey et al, (1996) J.Mol.Biol., 261 , 470-489) or ICM (Abagyan et al, (1994) J.Comput.Chem., 15, 488-506). This procedure can include computer fitting of binding partners to the adenosine A_¾ receptor structure to ascertain how well the shape and the chemical structure of the binding partner will bind to an adenosine AJJA receptor.

Thus the invention includes a method for the analysis of the interaction of one or more binding partners with adenosine A_¾ receptor comprising (a) constructing a computer representation of a binding region of the adenosine A2A receptor as defined by the coordinates of the human adenosine A_¾ receptor of Table (i), optionally varied by a root mean square deviation of residue backbone atoms of not more than 1.08 A, or selected coordinates thereof (b) selecting a binding partner to be evaluated by a method selected from the group consisting of assembling said binding partner; selecting a binding partner from a small molecule database; de novo ligand design of the binding partner; and modifying a known agonist or inhibitor, or a portion thereof, of an adenosine A_¾ receptor or homologue thereof; (c) employing computational means to dock said binding partner to be evaluated in a binding region in order to provide an energy-minimized configuration of the binding partner in a binding region; and (d) evaluating the results of said docking to quantify the interaction energy between said binding partner and the binding region.

Also computer-assisted, manual examination of the binding region structure of the adenosine A2A receptor may be performed. The use of programs such as GRID (Goodford, (1985) J. Med. Chem., 28, 849- 857) - a program that determines probable interaction sites between molecules with various functional groups and an enzyme surface - may also be used to analyse a binding region to predict, for example, the types of modifications which will alter the rate of metabolism of a binding partner.

Computer programs can be employed to estimate the attraction, repulsion, and steric hindrance of the adenosine A¾ receptor structure and a binding partner.

If more than one adenosine A2A receptor binding region is characterized and a plurality of respective smaller molecular fragments are designed or selected, a binding partner may be formed by linking the respective small molecular fragments into a single binding partner, which maintains the relative positions and orientations of the respective small molecular fragments at the binding sites. The single larger binding partner may be formed as a real molecule or by computer modelling. Detailed structural information can then be obtained about the binding of the binding partner to adenosine A¾ receptor, and in the light of this information adjustments can be made to the structure or functionality of the binding partner, e.g. to alter its interaction with adenosine A∑A receptor. The above steps may be repeated and re- repeated as necessary.

Thus, the three dimensional structural representation of the one or more binding partners of the third and fourth aspects of the invention may be obtained by: providing structural representations of a plurality of molecular fragments; fitting the structural representation of each of the molecular fragments to the coordinates of the human adenosine A2A receptor structural representation of Table (i), optionally varied by a root mean square deviation of residue C-a atoms of not more than 1.08 A, or selected coordinates thereof; and assembling the representations of the molecular fragments into one or more representations of single molecules to provide the three-dimensional structural representation of one or more candidate binding partners.

Typically the binding partner or molecule fragment is fitted to at least 5 or 10 non-hydrogen atoms of the adenosine A¾ receptor structure, preferably at least 20, 30, 40, 50, 60, 70, 80 or 90 non-hydrogen atoms and more preferably at least 100, 150, 200, 250, 300, 350, 400, 450, or 500 non-hydrogen atoms.

The invention includes screening methods to identify drugs or lead compounds of use in treating a disease or condition. For example, large numbers of binding partners, for example in a chemical database, can be screened for their ability to bind to adenosine A2A receptor. It is appreciated that in the methods described herein, which may be drug screening methods, a term well known to those skilled in the art, the binding partner may be a drug-like compound or lead compound for the development of a drug-like compound. The term "drug-like compound" is well known to those skilled in the art, and may include the meaning of a compound that has characteristics that may make it suitable for use in medicine, for example as the active ingredient in a medicament. Thus, for example, a druglike compound may be a molecule that may be synthesised by the techniques of organic chemistry, less preferably by techniques of molecular biology or biochemistry, and is preferably a small molecule, which may be of less than 1000 daltons (such as less than 500 daltons) and which may be water-soluble. A drug-like compound may additionally exhibit features of selective interaction with a particular protein or proteins and be bioavailable and/or able to penetrate target cellular membranes or the blood:brain barrier, but it will be appreciated that these features are not essential.

The term "lead compound" is similarly well known to those skilled in the art, and may include the meaning that the compound, whilst not itself suitable for use as a drug (for example because it is only weakly potent against its intended target, non-selective in its action, unstable, poorly soluble, difficult to synthesise or has poor bioavailability) may provide a starting-point for the design of other compounds that may have more desirable characteristics.

Thus in one embodiment of the methods of third and fourth aspects of the invention, the methods further comprise modifying the structural representation of the binding partner so as to increase or decrease their interaction with adenosine Aa_, receptor.

For example, once a binding partner has been designed or selected by the above methods, the efficiency with which that binding partner may bind to an adenosine Α_¾_, receptor may be tested and optimised, for example by computational evaluation. For example, a binding partner designed or selected as binding to an adenosine A2A receptor may be further computationally optimised so that in its bound state it would preferably lack repulsive electrostatic interaction with the target adenosine A₂A receptor and with the surrounding water molecules. Such non-complementary electrostatic interactions include repulsive charge-charge, dipole-dipole and charge-dipole interactions. Furthermore, it is often desired that binding partners demonstrate a relatively small difference in energy between the bound and free states (i.e., a small deformation energy of binding). Thus, binding partners may be designed with a deformation energy of binding of not greater than about 10 kcal/mole, more preferably, not greater than 7 kcal/mole. Binding partners may interact with the binding region in more than one conformation that is similar in overall binding energy. In those cases, the deformation energy of binding is taken to be the difference between the energy of the free binding partner and the average energy of the conformations observed when the binding partner binds to the protein. Specific computer software is available in the art to evaluate compound deformation energy and electrostatic interactions as detailed in WO2008/068534 (see, for example, page 34) incorporated herein by reference.

By modifying the structural representation we include, for example, 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 binding partner is changed while its original binding to adenosine A¾ receptor capability is increased or decreased. Such optimisation is regularly undertaken during drug development programmes to e.g. enhance potency, promote pharmacological acceptability, increase chemical stability etc. of lead compounds.

Examples of modifications include substitutions or removal of groups containing residues which interact with the amino acid side chain groups of the adenosine A_¾ receptor structure of the invention, as described further in relation to the β-adrenergic receptor in WO2008/068534 (see for example, page 35), incorporated herein by reference.

The potential binding effect of a binding partner on adenosine receptor may be analysed prior to its actual synthesis and testing by the use of computer modeling techniques. If the theoretical structure of the given entity suggests insufficient interaction and association between it and the adenosine A2A receptor, testing of the entity is obviated. However, if computer modelling indicates a strong interaction, the molecule may then be synthesized and tested for its ability to bind to an adenosine A∑A receptor. In this manner, synthesis of inoperative compounds may be avoided.

Thus in a further embodiment of the third and fourth aspects of the invention, the methods further comprise the steps of obtaining or synthesising the one or more binding partners of an adenosine A∑A receptor; and optionally contacting the one or more binding partners with an adenosine A2A receptor to determine the ability of the one or more binding partners to interact with the adenosine A2A receptor. Various methods known in the art may be used to determine binding between an adenosine A2A receptor and a binding partner including those described in WO2008/068534 (see for example, pages 35-36) incorporated herein by reference.

Once computer modelling has indicated that a binding partner has a strong interaction, it is appreciated that it may be desirable to crystallise a complex of the adenosine A_¾ receptor with that binding partner and analyse its interaction further by X-ray crystallography.

Thus in a further embodiment of the third and fourth aspects of the invention, the methods further comprise the steps of obtaining or synthesising the one or more binding partners of an adenosine A¾ receptor; forming one or more complexes of the adenosine A¾ receptor and the one or more binding partners; and analysing the one or more complexes by X-ray crystallography to determine the ability of the one or more binding partners to interact with adenosine A¾ receptor. Thus, it will be appreciated that another particularly useful drug design technique enabled by this invention is iterative drug design. Iterative drug design is a method for optimizing associations between a protein and a binding partner by determining and evaluating the three-dimensional structures of successive sets of protein/compound complexes, and is described further in WO2008/068534 (see, for example, pages 36 - 37), incorporated herein by reference.

The ability of a binding partner to modify adenosine A₂A receptor function may also be tested. For example the ability of a binding partner to modulate an adenosine A₂A receptor function could be tested by a number of well known standard methods, described extensively in the prior art.

In addition to in silico analysis and design, the interaction of one or more binding partners with an adenosine A2A receptor may be analysed directly by X-ray crystallography experiments, wherein the coordinates of the human adenosine A₂A receptor of Table (i), optionally varied by a root mean square deviation of residue backbone atoms of not more than 1.08 A, or selected coordinates thereof, are used to analyse the a crystal complex of the adenosine A2A receptor and binding partner. This can provide high resolution information of the interaction and can also provide insights into a mechanism by which a binding partner exerts an agonistic or antagonistic function. Accordingly, a fifth aspect of the invention provides a method for the analysis of the interaction of one or more binding partners with adenosine A2A receptor, comprising: obtaining or synthesising one or more binding partners; forming one or more crystallised complexes of an adenosine Aa_, receptor and a binding partner; and analysing the one or more complexes by X-ray crystallography by employing the coordinates of the human adenosine A_¾ receptor structure, of Table (i), optionally varied by a root mean square deviation of residue backbone atoms of not more than 1.08 A, or selected coordinates thereof, to determine the ability of the one or more binding partners to interact with the adenosine A2A receptor. Preferences for the selected coordinates in this and all subsequent aspects of the invention are as defined above with respect to the first aspect of the invention.

The analysis of such structures may employ X-ray crystallographic diffraction data from the complex and the coordinates of the human adenosine A2A receptor structure, of Table (i), optionally varied by a root mean square deviation of residue backbone atoms of not more than 1.08 A, or selected coordinates thereof, to generate a difference Fourier electron density map of the complex. The difference Fourier electron density map may then be analysed. In one embodiment, the one or more crystallised complexes are formed by soaking a crystal of adenosine A¾ receptor with the binding partner to form a complex. Alternatively, the complexes may be obtained by cocrystallising the adenosine A_¾ receptor with the binding partner. For example a purified adenosine A_¾ receptor protein sample is incubated over a period of time (usually >1 hr) with a potential binding partner and the complex can then be screened for crystallization conditions. Alternatively, protein crystals containing a first binding partner can be back-soaked to remove this binding partner by placing the crystals into a stabilising solution in which the binding partner is not present. The resultant crystals can then be transferred into a second solution containing a second binding partner and used to produce an X-ray diffraction pattern of adenosine A_2A receptor complexed with the second binding partner. The complexes can be analysed 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 adenosine A2A receptor and the solved structure of uncomplexed adenosine A2A receptor . This is described further in WO2008/068534 (see, for example, pages 38 - 39), incorporated herein by reference.

This information may thus be used to optimise known classes of adenosine A^ receptor binding partners and to design and synthesize novel classes of adenosine A_¾ receptor binding partners, particularly those which have agonistic or antagonistic properties, and to design drugs with modified adenosine A_¾ receptor interactions.

In one approach, the structure of a binding partner bound to an adenosine A2A receptor may be determined by experiment. This will provide a starting point in the analysis of the binding partner bound to adenosine A_¾ receptor thus providing those of skill in the art with a detailed insight as to how that particular binding partner interacts with adenosine A2A receptor and the mechanism by which it exerts any function effect.

Many of the techniques and approaches applied to structure-based drug design described above rely at some stage on X-ray analysis to identify the binding position of a binding partner 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 binding partner. 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 three dimensional structure (or at least a set of structure factors for the protein crystal). Therefore, determination of the adenosine A_¾ receptor structure also allows difference Fourier electron density maps of adenosine A_¾ receptor -binding partner complexes to be produced, determination of the binding position of the binding partner and hence may greatly assist the process of rational drug design.

Accordingly, a sixth aspect of the invention provides a method for predicting the three dimensional structure of a binding partner of unknown structure, or part thereof, which binds to adenosine A2A receptor, comprising: providing the coordinates of the adenosine A2A receptor structure of Table (i), optionally varied by a root mean square deviation of residue backbone atoms of not more than 1.08 A, or selected coordinates thereof; providing an X-ray diffraction pattern of adenosine A∑A receptor complexed with the binding partner; and using the coordinates to predict at least part of the structure coordinates of the binding partner.

In one embodiment, the X-ray diffraction pattern is obtained from a crystal formed by soaking a crystal of adenosine A2A receptor with the binding partner to form a complex. Alternatively, the X-ray diffraction pattern is obtained from a crystal formed by cocrystallising the adenosine A2A receptor with the binding partner as described above. Alternatively, protein crystals containing a first binding partner can be back-soaked to remove this binding partner and the resultant crystals transferred into a second solution containing a second binding partner as described above.

A mixture of compounds may be soaked or co-crystallized with an adenosine A2A receptor crystal, wherein only one or some of the compounds may be expected to bind to the adenosine A_¾ receptor. The mixture of compounds may comprise a ligand known to bind to adenosine A_M receptor. As well as the structure of the complex, the identity of the complexing compound(s) is/are then determined.

Preferably, the methods of the previous aspects of the invention are computer-based. For example, typically the methods of the previous aspects of the invention make use of the computer systems and computer-readable storage mediums of the ninth and tenth aspects of the invention.

A seventh aspect of the invention provides a method for producing a binding partner of adenosine A2A receptor comprising: identifying a binding partner according to the third, fourth, fifth or sixth aspects of the invention and synthesising the binding partner.

The binding partner may be synthesised using any suitable technique known in the art including, for example, the techniques of synthetic chemistry, organic chemistry and molecular biology.

It will be appreciated that it may be desirable to test the binding partner in an in vivo or in vitro biological system in order to determine its binding and/or activity and/or its effectiveness. For example, its binding to an adenosine A2A receptor may be assessed using any suitable binding assay known in the art including the examples described above. Alternatively, is ability to modulate the adenosine A2A receptor's ability to form dimers may be assessed. Moreover, its effect on adenosine A2A receptor function in an in vivo or in vitro assay may be tested. For example, the effect of the binding partner on the adenosine A2A receptor signalling pathway may be determined. For example, the activity may be measured by using a reporter polynucleotide to measure the activity of the adenosine A2A receptor signalling pathway. By a reporter polynucleotide we include genes which encode a reporter protein whose activity may easily be assayed, for example β-galactosidase, chloramphenicol acetyl transferase (CAT) gene, luciferase or Green Fluorescent Protein (see, for example, Tan ef al, 1996 EMBO J 15(17): 4629-42). Several techniques are available in the art to detect and measure expression of a reporter polynucleotide which would be suitable for use in the present invention. Many of these are available in kits both for determining expression in vitro and in vivo. Alternatively, signalling may be assayed by the analysis of downstream targets. For example, a particular protein whose expression is known to be under the control of a specific signalling pathway may be quantified. Protein levels in biological samples can be determined using any suitable method known in the art. For example, protein concentration can be studied by a range of antibody based methods including immunoassays, such as ELISAs, western blotting and radioimmunoassays.

An eight aspect of the invention provides a binding partner produced by the method of the seventh aspect of the invention.

Following identification of a binding partner, it may be manufactured and/or used in the preparation, i.e. manufacture or formulation, of a composition such as a medicament, pharmaceutical composition or drug. These may be administered to individuals.

Accordingly, the invention includes a method for producing a medicament, pharmaceutical composition or drug, the process comprising: (a) providing a binding partner according to the eighth aspect of the invention and (b) preparing a medicament, pharmaceutical composition or drug containing the binding partner.

The medicaments may be used to treat any disorder or condition ameliorated by modulation of the A2A receptor. Examples include: heart failure (such as acute decompensated heart failure and congestive heart failure); kidney failure (e.g. caused by heart failure); oedema; cancer (such as prostate, rectal, renal, ovarian, endometrial, thyroid, pancreatic, particularly breast, colon, bladder, brain, glia, melanoma, pineal gland and, more particularly, lung cancer (e.g. Lewis lung carcinoma)); diabetes; diarrhea; macular degeneration (such as macular degeneration caused by angiogenesis (e.g. retinal angiogenesis)); or, particularly (e.g. for disorders or conditions ameliorated by the inhibition of the A_2a receptor), a disease of the central nervous system such as depression, a cognitive function disease, a neurodegenerative disease (such as Parkinson's disease, Huntington's disease, Alzheimer's disease, amyotrophic lateral sclerosis) and psychoses; an attention related disorder (such as attention deficit disorder (ADD) and attention deficit hyperactivity disorder (ADHD)); extra pyramidal syndrome (e.g. dystonia, akathisia, pseudoparkinsonism and tardive dyskinesia); a disorder of abnormal movement (such as restless leg syndrome (RLS) and periodic limb movement in sleep (PLMS)); cirrhosis; liver fibrosis; fatty liver; dermal fibrosis (e.g. in diseases such as scleroderma); a sleep disorder; stroke; brain injury and neuroinflammation (e.g. migraine or any disorder or condition caused by ischemia, stroke, head injury or CNS inflammation); addictive behaviour.

The invention also provides systems, particularly a computer system, intended to generate structures and/or perform optimisation of binding partner which interact with adenosine A_¾ receptor, adenosine A_M receptor homologues or analogues, complexes of adenosine A2A receptor with binding partners, or complexes of adenosine A_¾ receptor homologues or analogues with binding partners. Accordingly, a ninth aspect of the invention provides a computer system, intended to generate three dimensional structural representations of adenosine A^ receptor, adenosine A2A receptor homologues or analogues, complexes of adenosine A2A receptor with binding partners, or complexes of adenosine A_¾ receptor homologues or analogues with binding partners, or, to analyse or optimise binding of binding partners to said adenosine A_¾ receptor or homologues or analogues, or complexes thereof, the system containing computer-readable data comprising one or more of:

(a) the coordinates of the adenosine A2A receptor structure of Table (i), optionally varied by a root mean square deviation of residue backbone atoms of not more than 1.08 A, or selected coordinates thereof;

(b) the coordinates of a target adenosine A₂A receptor homologue or analogue generated by homology modelling of the target based on the data in (a);

(c) the coordinates of a binding partner generated by interpreting X-ray crystallographic data or NMR data by reference to the coordinates of the adenosine A2A receptor structure, of Table (i), optionally varied by a root mean square deviation of residue backbone atoms of not more than 1.08 A, or selected coordinates thereof, and

(d) structure factor data derivable from the coordinates of (a), (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 drug design. The computer system may further comprise a display coupled to the central-processing unit for displaying structural representations.

The invention also provides such systems containing atomic coordinate data of target proteins of unknown structure wherein such data has been generated according to the methods of the invention described herein based on the starting data provided in Table (i), optionally varied by a root mean square deviation of residue backbone atoms of not more than 1.08 A, or selected coordinates thereof.

Such data is useful for a number of purposes, including the generation of structures to analyse the mechanisms of action of binding partners and/or to perform rational drug design of binding partners which interact with adenosine A_¾ receptors, such as compounds which are agonists or antagonists.

A tenth aspect of the invention provides a computer-readable storage medium, comprising a data storage material encoded with computer readable data, wherein the data comprises one or more of:

(a) the coordinates of the human adenosine A2A receptor structure of Table (i), optionally varied by a root mean square deviation of residue backbone atoms of not more than 1.08 A, or selected coordinates thereof;

(b) the coordinates of a target adenosine A2A receptor homologue or analogue generated by homology modelling of the target based on the data in (a);

(c) the coordinates of a binding partner generated by interpreting X-ray crystallographic data or NMR data by reference to the coordinates of the adenosine A2A receptor structure of Table (i), optionally varied by a root mean square deviation of residue backbone atoms of not more than 1.08 A, or selected coordinates thereof, and

(d) structure factor data derivable from the coordinates of (a), (b) or (c). The invention also includes a computer-readable storage medium comprising a data storage material encoded with a first set of computer-readable data comprising a Fourier transform of at least a portion of the structural coordinates of adenosine A∑A receptor, of Table (i), optionally varied by a root mean square deviation of residue backbone atoms of not more than 1.08 A, or selected coordinates thereof; which data, 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 e.g. a target protein 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 structure coordinates corresponding to the second set of machine readable data.

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 structural coordinates of adenosine A2A receptor, of Table (i), optionally varied by a root mean square deviation of residue backbone atoms of not more than1.08 A, 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, e.g. a target protein 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.

It will be appreciated the that the computer-readable storage media of the invention may comprise a data storage material encoded with any of the data generated by carrying out any of the methods of the invention relating to structure solution and selection/design of binding partners to adenosine A2A receptor and drug design.

The invention also includes a method of preparing the computer-readable storage media of the invention comprising encoding a data storage material with the computer-readable data. 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 adenosine A2A receptor 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 analyse 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 XP or IBM OS/2 operating systems.

An eleventh aspect of the invention provides a method for providing data for generating three dimensional structural representations of adenosine A_¾ receptor, adenosine A_¾ receptor homologues or analogues, complexes of adenosine A∑A receptor with binding partners, or complexes of adenosine A2A receptor homologues or analogues with binding partners, or, for analysing or optimising binding of binding partners to said adenosine A_¾ receptor or homologues or analogues, or complexes thereof, the method comprising:

(i) establishing communication with a remote device containing computer-readable data comprising at least one of:

(a) the coordinates of the human adenosine A_¾ receptor structure of Table (i), optionally varied by a root mean square deviation of residue backbone atoms of not more than 1.08 A, or selected coordinates thereof;

(b) the coordinates of a target adenosine A_¾ receptor homologue or analogue generated by homology modelling of the target based on the data in

(a) ;

(c) the coordinates of a binding partner generated by interpreting X-ray crystallographic data or NMR data by reference to the coordinates of the human adenosine A2A receptor structure of Table (i), optionally varied by a root mean square deviation of residue backbone atoms of not more than1.08 A, or selected coordinates thereof, and

(d) structure factor data derivable from the coordinates of (a),

(b) or (c); and (ii) receiving said computer-readable data from said remote device.

The computer-readable data received from said remote device, particularly when in the form of the coordinates of the adenosine A¾ receptor structure of Table (i), optionally varied by a root mean square deviation of residue backbone atoms of not more than 1.08 A, or selected coordinates thereof, may be used in the methods of the invention described herein, e.g. for the analysis of a binding partner structure with an adenosine A_∑A receptor structure.

Thus the remote device may comprise e.g. a computer system or computer readable media of one of the previous aspects of the invention. The device may be in a different country or jurisdiction from where the computer-readable data is received.

The communication may be via the internet, intranet, e-mail etc, transmitted through wires or by wireless means such as by terrestrial radio or by satellite. Typically the communication will be electronic in nature, but some or all of the communication pathway may be optical, for example, over optical fibers.

A twelfth aspect of the invention provides a method of obtaining a three dimensional structural representation of a crystal of an adenosine A2A receptor, which method comprises providing the coordinates of the human adenosine A_¾ receptor structure of Table (i), optionally varied by a root mean square deviation of residue backbone atoms of not more than 1.08 A, or selected coordinates thereof, and generating a three-dimensional structural representation of said coordinates. For example, the structural representation may be a physical representation or a computer generated representation. Examples of representations are described above and include, for example, any of a wire-frame model, a chicken-wire model, a ball-and-stick model, a spacefilling model, a stick model, a ribbon model, a snake model, an arrow and cylinder model, an electron density map or a molecular surface model.

Computer representations can be generated or displayed by commercially available software programs including for example QUANTA (Accelrys .COPYRIGHT.2001 , 2002), O (Jones et al., Acta Crystallogr. A47, pp. 110-119 (1991)) and RIBBONS (Carson, J. Appl. Crystallogr., 24, pp. 9589-961 (1991)). Typically, the computer used to generate the representation comprises (i) a computer- readable data storage medium comprising a data storage material encoded with computer- readable data, wherein said data comprise the coordinates of the adenosine A2A receptor structure, of Table (i), optionally varied by a root mean square deviation of residue backbone atoms of not more than 1.08 A, or selected coordinates thereof; and (ii) instructions for processing the computer-readable data into a three-dimensional structural representation. The computer may further comprise a display for displaying said three-dimensional representation. A thirteenth aspect of the invention provides a method of predicting one or more sites of interaction of an adenosine A2A receptor or a homologue thereof, the method comprising: providing the coordinates of the human adenosine A2A receptor structure of Table (i), optionally varied by a root mean square deviation of residue backbone atoms of not more than 1.08 A, or selected coordinates thereof; and analysing said coordinates to predict one or more sites of interaction.

For example, a binding region of an adenosine A2A receptor for a particular binding partner can be predicted by modelling where the structure of the binding partner is known. Typically, the fitting and docking methods described above would be used. This method may be used, for example, to predict the site of interaction of a G protein of known structure as described in viz Gray J J (2006) Curr Op Struc Biol Vol 16, pp 183-193.

A fourteenth aspect of the invention provides a method for assessing the activation state of a structure for adenosine A_¾ receptor, comprising: providing the coordinates of the human adenosine A_¾ receptor structure, of Table (i), optionally varied by a root mean square deviation of residue backbone atoms of not more than 1.08 A, or selected coordinates thereof; performing a statistical and/or topological analysis of the coordinates; and comparing the results of the analysis with the results of an analysis of coordinates of proteins of known activation states.

For example, protein structures may be compared for similarity by statistical and/or topological analyses (suitable analyses are known in the art and include, for example those described in Grindley et al (1993) J Mol Biol Vol 229: 707-721 and Holm & Sander (1997) Nucl Acids Res Vol 25: 231-234). Highly similar scores would indicate a shared conformational and therefore functional state eg the inactive antagonist state in this case. One example of statistical analysis is multivariate analysis which is well known in the art and can be done using techniques including principal components analysis, hierarchical cluster analysis, genetic algorithms and neural networks. By performing a multivariate analysis of the coordinate data of the adenosine A_¾ receptor structure of Table (i), optionally varied by a root mean square deviation of residue backbone atoms of not more than 1.08 A, or selected coordinates thereof, and comparing the result of the analysis with the results of the analysis performed on coordinates of proteins with known activation states, it is possible to determine the activation state of the coordinate set analysed. For example, the activation state may be classified as 'active' or 'inactive'.

A fifteenth aspect of the invention provides a method of producing a protein with a binding region that has substrate specificity substantially identical to that of adenosine A_¾ receptor, the method comprising:

a) aligning the amino acid sequence of a target protein with the amino acid sequence of an adenosine A2A receptor;

b) identifying the amino acid residues in the target protein that contribute to the adenosine binding site of the A_¾ receptor and correspond to any one or more of the following positions according to the numbering of the adenosine A2A receptor as set out in Figure 13: Ile66, Ala63, lie 274, Val84, His278, Ser277, Leu249, Trp246, Thr88, Asn181, His250, Met177, Asn253, Met 270, Glu169 and Phe168; or identifying the amino acid residues in the target protein that that contribute to the NECA binding site of the A_¾ receptor and correspond to any one or more of the following positions according to the numbering of the adenosine A2A receptor as set out in Figure 13: Ile66, Ala63, lie 274, Val84, His278, Ser277, Leu249, Trp246, Thr88, Asn181 , His250, Met177, Asn253, Met 270, Glu169, Phe168, Leu85 and Tyr9; and

c) making one or more mutations in the amino acid sequence of the target protein to replace one or more identified amino acid residues with the corresponding residue in the adenosine A2A receptor .

By "an amino acid residue that corresponds to" we include an amino acid residue that aligns to the given amino acid residue in adenosine A_2A receptor when the adenosine A₂A receptor and target protein are aligned using e.g. MacVector and CLUSTALW.

For example, amino acid residues contributing to the adenosine binding site include amino acid residues Ile66, Ala63, lie 274, Val84, His278, Ser277, Leu249, Trp246, Thr88, Asn 81 , His250, Met177, Asn253, Met 270, Glu169 and Phe168 or amino acid residues contributing to the NECA binding site include amino acid residues Ile66, Ala63, lie 274, Val84, His278, Ser277, Leu249, Trp246, Thr88, Asn181 , His250, Met177, Asn253, Met 270, Glu169, Phe168, Leu85 and Tyr9. Thus a binding site of a particular protein may be engineered using well known molecular biology techniques to contain any one or more of these residues to give it the same substrate specificity. This technique is well known in the art and is described in, for example, Ikuta et al (J Biol Chem (2001) 276, 27548-27554) where the authors modified the active site of cdk2, for which they could obtain structural data, to resemble that of cdk4, for which no X-ray structure was available.

Preferably, all 18 amino acids in the target portion which correspond to amino acid residues Ile66, Ala63, lie 274, Val84, His278, Ser277, Leu249, Trp246, Thr88, Asn181 , His250, Met177, Asn253, Met 270, Glu169, Phe168, Leu85 and Tyr9 of the adenosine A_ZA receptor are, if different, replaced. However, it will be appreciated that only 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2 or 1 amino acid residues may be replaced.

Preferences for the target protein are as defined above with respect to the first aspect of the invention. A sixteenth aspect of the invention provides a method of predicting the location of internal and/or external parts of the structure of adenosine A_¾ receptor or a homologue thereof, the method comprising: providing the coordinates of the adenosine A2A receptor structure of Table (i), optionally varied by a root mean square deviation of residue backbone atoms of not more than 1.08 A, or selected coordinates thereof and analysing said coordinates to predict the location of internal and/or external parts of the structure.

For example, from the three dimensional representation, it is possible to read off external parts of the structure, eg surface residues, as well as internal parts, eg residues within the protein core. It will be appreciated that the identification of external protein sequences will be especially useful in the generation of antibodies against an adenosine A_2A receptor.

The crystallisation of the adenosine A2A receptor has led to many interesting observations about its structure, including its adenosine- and NEC-binding sites. Thus it will be appreciated that the invention allows for the generation of mutant adenosine A_2A receptors wherein residues corresponding to these areas of interest are mutated to determine their effect on adenosine A2A receptor function, ligand binding specificity, and dimerisation capability.

Accordingly, a seventeenth aspect of the invention provides a mutant adenosine A_¾ receptor which, when compared to the corresponding wild-type adenosine receptor, has a different amino acid at a position which corresponds to any one or more of the following positions according to the numbering of the human adenosine A2A receptor as set out in Figure 13: Ile66, Ala63, lie 274, Val84, His278, Ser277, Leu249, Trp246, Thr88, Asn181 , His250, Met177, Asn253, Met 270, Glu169, Phe168, Leu85 and Tyr9. As mentioned above, these amino acids define the adenosine binding site in the human adenosine A_M receptor.

The invention also provides a mutant adenosine A_¾ receptor which, when compared to the corresponding wild-type adenosine A_¾ receptor, has a different amino acid at a position which corresponds to any one or more of the following positions according to the numbering of the human adenosine A_M receptor as set out in Figure 13: Ile66, Ala63, lie 274, Val84, His278, Ser277, Leu249, Trp246, Thr88, Asn181 , His250, Met177, Asn253, Met 270, Glu169, Phe168, Leu85 and Tyr9. As mentioned above, these amino acids define the NECA binding site in the human adenosine A_¾ receptor. It is particularly preferred if the mutant adenosine receptor of the invention is one which has at least 20% amino acid sequence identity when compared to the given human adenosine A2A receptor, as determined using MacVector and CLUSTALW. Preferably, the mutant adenosine receptor has at least 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 95% or 99% amino acid sequence identity.

The mutant adenosine receptor may be a mutant of any adenosine receptor provided that it is mutated at one or more of the amino acid positions as stated by reference to the given human adenosine A2A receptor amino acid sequence. Thus, the invention includes a mutant human adenosine A_¾ receptor in which, compared to its parent, one or more of these amino acid residues have been replaced by another amino acid residue. The invention also includes mutant adenosine receptors from other sources in which one or more corresponding amino acids in the parent receptor are replaced by another amino acid residue. For the avoidance of doubt the parent may be an adenosine receptor which has a naturally-occurring sequence, or it may be a truncated form or it may be a fusion, either to the naturally-occurring protein or to a fragment thereof, or it may contain mutations compared to the naturally-occurring sequence, providing that it retains ligand-binding ability.

In an embodiment, the mutant adenosine receptor of the invention has a combination of 2 or 3 or 4 or 5 or 6 or 7 or 8 or 9 or 10 mutations as described above.

By "corresponding amino acid residue" we include the meaning of the amino acid residue in another adenosine receptor which aligns to the given amino acid residue in the human adenosine A2A receptor when the human adenosine A_¾ receptor and the other adenosine receptor are compared using MacVector and CLUSTALW.

Residues in proteins can be mutated using standard molecular biology techniques as are well known in the art. An eighteenth aspect of the invention provides a method of making an adenosine A_M receptor crystal comprising: providing purified adenosine A_M receptor; and crystallising the adenosine A_¾ receptor either by using the sitting drop or hanging drop vapour diffusion technique, using a precipitant solution comprising 0.05 M ADA-NaOH, pH 6.4, 23.6% PEG 400 , 4% v/v 2-propanol. Any PEG from PEG200 to PEG5000 may be used, such as from PEG200 to PEG1500 (e.g. PEG 1000).

In a particularly preferred embodiment, the precipitant solution 0.05 M ADA-NaOH, pH 6.4, 23.6% PEG 400, 4% v/v 2-propanol and cholesteryl hemisuccinate (CHS) (0.1mg/ml) and 0.5% OTG (n-octyl beta-D-thioglucopyranoside at 4°C.

In a further embodiment, the precipitant solution comprises 20-30% PEG400.

A nineteenth aspect of the invention provides a crystal of adenosine A2A receptor having the structure defined by the coordinates of the human adenosine A2A receptor structure of Table (i), optionally varied by a root mean square deviation of residue backbone atoms of not more than 1.08 A, or selected coordinates thereof. Typically, the crystal has a resolution of 3.0 A for the adenosine-bound structure and 2.6 A for the NECA-bound structure.

The space group of the crystal C121. Thus, in one embodiment the crystal has C2 symmetry with unit cell dimensions a=76.9 (± 10) A, b=99.6 (± 10) A, c=79.7 (± 10) A, wherein a= 90 β = 93.3 (± 10)° and γ = 90 for NECA; a=76.5 (± 15) A, b=98.9 (± 15) A, c=79.5 (± 15) A, wherein a= 90 )°, β = 93.5 (± 10)° and γ = 90 (± 10)°.

The invention also includes a co-crystal of adenosine A^ receptor having the structure defined by the coordinates of the adenosine A_¾ receptor structure of Table (i), optionally varied by a root mean square deviation of residue backbone atoms of not more than 1.08 A, or selected coordinates thereof, and a binding partner. Typically, the crystal has a resolution of 3.0 A for the adenosine-bound structure and 2.6 A for the NECA-bound structure.

The invention includes the use of the coordinates of the human adenosine A_¾ receptor structure of Table (i), optionally varied by a root mean square deviation of residue backbone atoms of not more than 1.08 A, or selected coordinates thereof to solve the structure of target proteins of unknown structure.

The invention includes the use of the coordinates of the adenosine A_¾ receptor structure of Table (i), optionally varied by a root mean square deviation of residue backbone atoms of not more than 1.08 A, or selected coordinates thereof to identify binding partners of an adenosine A_¾ receptor.

The invention includes the use of the coordinates of the adenosine A2A receptor structure of Table (i), optionally varied by a root mean square deviation of residue backbone atoms of not more than 1.08 A, or selected coordinates thereof in methods of drug design where the drugs are aimed at modifying the activity of the adenosine A2A receptor.

A twentieth aspect of the invention provides a method of predicting a three dimensional structural representation of an active state of a target protein, or part thereof, comprising:

predicting the three-dimensional structural representation of the active state of the target protein, or part thereof, by modelling a structural representation of the target protein in a non-active state on all or the selected coordinates of the adenosine A2A receptor. Preferably the active state is a fully active state. However, the active state may be the partially active state described in relation to A2A-GL31 in the Examples section, below (i.e. an intermediate conformation between the inactive and active states). Preferably the non-active state is the fully inactive ground state.

A twenty-first aspect of the invention provides a method of predicting a three dimensional structural representation of an inactive state of a target protein, or part thereof, comprising:

predicting the three-dimensional structural representation of the inactive state of the target protein, or part thereof, by modelling a structural representation of the target protein in an active state on all or the selected coordinates of the adenosine A¾ receptor.

Preferably the active state is a fully active state. However, the active state may be the partially active state described in relation to A2A-GL31 in the Examples section, below (i.e. an intermediate conformation between the inactive and active states).

Preferably the non-active state is the fully inactive ground state.

The invention will now be described in more detail with the aid of the following Figures and Examples.

Figure 1: Structure of the adenosine A_¾ receptor bound to NECA compared to other GPCR structures, (a) The structure of NECA-bound A^R is shown as a cartoon (yellow) aligned with the structure of A2A-T4L bound to the inverse agonist ZM241385 (blue; PDB code 3EML⁸). NECA is shown as a space-filling model (C, green; N, blue; O, red), (b, c) Sections through the aligned receptors in (a) to highlight the differences in the intracellular face of the receptors (b) and in the ligand binding pocket (c), with the bulge in H5 shown as an inset, (d, e) Alignment of NECA-bound A2AR (yellow) with agonist-bound beta₂AR-Nb80 (red; PDB code 3P0G⁵) showing the intracellular face of the receptors (d) and the ligand binding pocket (e). NECA is shown as a space-filling model in c and e. The figures were generated using CCP4mg³¹. Analogous alignments to opsin are depicted in Fig. 11. Figure 2: Receptor-ligand interactions compared for the adenosine receptor bound to the inverse agonist ZM241385 and the agonists NECA and adenosine. Structures of the human A2AR in cartoon representation are shown bound to the following ligands: (a) ZM241385 (PDB code 3EML⁸); (b) NECA; (c) adenosine, (d, e) Polar and non-polar interactions involved in agonist binding to A_¾R are shown for NECA (d) and adenosine (e). Amino acid residues within 3.9 A of the ligands are depicted, with residues highlighted in blue making van der Waals contacts (blue rays) and residues highlighted in red making potential hydrogen bonds with favourable geometry (red dashed lines, as identified by HBPLUS, see Methods) or hydrogen bonds with unfavourable geometry (blue dashed lines, donor acceptor distance less than 3.6 A). Where the amino acid residue differs between the human A2AR and the human A^R, A₂BR and A₃R, the equivalent residue is shown highlighted in orange, purple or green, respectively. Panels a-c were generated using Pymol (DeLano Scientific Ltd). Omit densities for the ligands are shown in Fig. 10 and densities for water molecules in Fig. 12. (d) Amino acid residues highlighted in blue: Ala 63, He 66, Phe 168, Met 270, lie 274, Val 84, Met 177, Leu 249, Trp 246, Leu 85, Asn 181 ; amino acid residues highlighted in red: Glu 169, Thr 256, Asn 253, His 278, Ser 277, Thr 88, Tyr 9, His 250; amino acid residues highlighted in green: Val 72, Val 169, Ser 247; amino acid residue highlighted in orange: Thr 277; amino acid residue highlighted in purple: Val 250. (e) Amino acid residues highlighted in blue: Ala 63, lie 66, Phe 168, Met 270, He 274, Val 84, Met 177, Leu 249, Trp 246, Thr 88; amino acid residues highlighted in red: Glu 169, Asn 253, His 278, Ser 277, His 250, Asn 181 ; amino acid residues highlighted in green: Val 72, Val 169, Ser 247; amino acid residue highlighted in orange: Thr 277.

Figure 3: Positions of adenosine and ZM241385 in the adenosine A2A receptor ligand binding pocket. The structures of adenosine-bound A2AR-GL3 and ZM241385-bound A2A- T4L were aligned using only atoms from the protein to allow the ligand positions to be compared, with adenosine in yellow and ZM241385 in pink (N, blue; O, red). The ligands are shown in the context of the binding pocket of A2AR-GL31 , with transmembrane helices of A2AR-GL3 shown in yellow and the surfaces of the receptor, including the cavity of the ligand binding pocket, shown in grey. The side chains of Val84 and Leu85 that interact with the ribose moiety of the agonist are shown in green.

Figure 4: Comparison of the positions of agonists in the binding pockets of the adenosine A2A receptor and a beta-adrenoceptor. (a) The structures of A2AR bound to adenosine and betatAR bound to isoprenaline (PDB code 2Y03)⁷ were aligned by superimposing equivalent atoms in the protein structure and the positions of both ligands shown as stick models with the carbon atoms in magenta (isoprenaline) or yellow (adenosine) nitrogen in blue and oxygen in red. The Az_,R structure is shown, with H5 and H7 as space-filling models (C, grey; N, blue; O, red) and the remainder of the structure as a cartoon (pale green). Some water molecules are shown as red spheres, hydrogen bonds as red dashed lines and the polar contacts as blue dashed lines. The orientation of the figure is identical to that shown in Fig. 2. (b) Structure of A_¾R bound to adenosine viewed from the extracellular surface, (c) Structure of beta^R bound to isoprenaline (PDB code 2Y03)⁷ viewed from the extracellular surface. In panels b and c, equivalent side chains in the respective structures that make contacts to both isoprenaline and adenosine in their respective receptors are shown as space-filling models and they have the following Ballesteros-Weinstein numbers (amino acid side chains are shown in parentheses for the AZAR and beta^R, respectively): 3.32 (V84, D121); 3.36 (T88, V125); 5.42 (N181*. S211); 6.51 (L249, F306); 6.55 (N253, N310); 6.52 (H250*, F307); 7.39 (I274, N329); 7.43 (H278, Y333). Some residues (*) only make indirect contacts to the agonists via a water molecule.

Figure 5: Competition binding curves for wild-type A2AR and A2AR-GL31 for adenosine, NECA and ZM241385. Ligand affinities (-log Ki values) for all ligands tested are given in the Table; *p<0.05; ***p<0.001 as compared using a one-way ANOVA with Dunnett's post-hoc test. All data are the mean ± s.e.m. of three experiments performed in duplicate (except adenosine at wild-type; n=2).

Figure 6: Effect of CGS21680 on cAMP accumulation in HEK293-TREx inducible cells expressing wildtype A2AR and A2AR-GL31 under the control of a doxycyline-induced promoter, (a) In the absence of doxycycline there is still a measurable agonist-mediated cAMP response at A2AR, suggesting 'leaky' receptor expression. In the presence of doxycycline (1 - 3 nM), the basal cAMP accumulation is elevated, indicative of increasing constitutive activity, to the point where no further agonist-mediated cAMP accumulation is observed, (b) For A2AR-GL31 , very similar levels of constitutive activity as seen for wildtype A2AR are observed in the presence of increasing doxycycline concentrations. However, CGS21680 is only poorly able to stimulate cAMP accumulation, indicating that the transmission of agonist-mediated receptor activation is impaired, (c) Overlay of fitted curves from A2AR and A2AR-GL31 to compare levels of constitutive and agonist-stimulated cAMP accumulation, (d) CGS21680-induced cAMP responses are expressed as a percentage of the maximum CGS21680 respose through the wild-type A2AR; parental HEK293cell line, black circles; wildtype A2AR, red squares; A2AR-GL31 , orange triangles. All data are the mean ± s.e.m. of three experiments performed in duplicate or triplicate. Figure 7: Alignment of A2AR-GL31 and opsin. The cytoplasmic surface of the aligned GPCRs is shown, with A2AR-GL31 in rainbow colouration (N-terminus blue, C-terminus red) and opsin in grey. The C-terminal peptide of the G protein transducin bound to the active state of opsin is shown (a) as a cartoon representation in purple or (b) as a space filling model (C, grey; O, red; N, blue). It is clear from (b) that there is insufficient room to fit the peptide in the structure of A2AR-GL31 unless there is an additional 3.5 A outward movement of the end of H6, assuming that Gas interacts with A2AR in a similar way to how transducin interacts with opsin. The alignment was performed using 'super align' in Pymol (Delano Scientific).

Figure 8: Local conformational re-arrangements in the environment of the thermostabilising mutations L48A (H2) and Q89A (H3). Backbone and side chains are represented in purple for the ZM241385-bound structure of A2A-T4L (PDB code 3EML) and in green for NECA- bound A2AR-GL31. In the ZM241385-bound structure, L48 is localized in the microenvironment of the highly conserved NPXXY motif (A). The mutations L48A in the A2AR-GL31 construct bound to NECA (B) induces reorganization of the hydrogen bond network involving H2 (L48A, D52) H7 (S281, P285) and H3 (S91). P285 (from the NPXXY motif) undergoes an isomerisation from the trans-conformation, as observed in the antagonist-bound state (C), to the cis- conformation in the agonist-bound conformation in A2AR-GL31 (D). The Q89 side chain is facing the bulge in TM5 as observed in the antagonist-bound state (E). Mutating the Q89 side chain, in Q89A, might be partly responsible for the inward movement of H5 which is also observed in the 2AR-Nb80 structure. The extracellular side is at the top of all the panels. Figure 9: Structures of A2AR ligands discussed in the main text. Parts of the structures in blue represent elements identical to those found in adenosine. In the structure of adenosine, the numbering system for carbon and nitrogen atoms is shown.

Figure 10: Fo-Fc omit map for NECA (a and b) and adenosine (c and d). Figures were made using Pymol and the contour level is 2.5 sigma. The extracellular surface is at the top of each figure.

Figure 11: Structure of A2AR-GL31 bound to NECA compared to that of opsin. The structure of NECA-bound A2AR-GL31 is shown as a cartoon (yellow) aligned with the structure of opsin complexed with a peptide derived from the C-terminus of Ga (PDB code 3DQB) in green. Sections through the receptors are shown located (a) at the intracellular face and (b) in the region of the ligand binding pocket of A2AR-GL31. NECA is shown as a space filling model (C, green; N, blue; O, red) in (b). The structural alignment was performed in the same manner as for the other receptors (see Online Methods). Opsin does not display the bulge in H5 observed in the agonist bound structures of A2AR-GL31 and P2AR-Nb80, but shows larger displacements of H5 and H6 than those found in the agonist bound A2AR-GL31. Figure generated with CCP4mg.

Figure 12: Electron density for water molecules located in the binding pockets of the agonist-bound A2AR structures. Sharpened (B = -15 A2) 2Fo-Fc electron density maps for the binding pockets of (a) NECA and (b) adenosine. The contour levels are 1.5σ in (a) and 1.2σ in (b). Waters are represented as red spheres. Figure generated with CCP4mg.

Figure 13: Amino acid sequence of the human A2A receptor as crystallised (SEQ ID No: 1). Figure 14: Amino acid sequence of wild type human A_¾ receptor (SEQ ID No: 2).

Example 1 : Agonist-bound adenosine A?_A receptor structures reveal common features of GPCR activation The adenosine receptors and beta-adrenoceptors (beta-ARs) are G protein-coupled receptors (GPCRs) that activate intracellular G proteins upon binding agonist such as adenosine¹ or noradrenaline², respectively. GPCRs have similar structures consisting of 7 transmembrane helices that contain well-conserved sequence motifs, suggesting that they are probably activated by a common mechanism^3,4. Recent structures of beta-ARs highlight residues in transmembrane region 5 that initially bind specifically to agonists rather than to antagonists, suggesting an important role in agonist-induced activation of receptors^5"7. Here we present two crystal structures of the thermostabilised human adenosine A_¾ receptor (A2AR-GL31) bound to its endogenous agonist adenosine and the synthetic agonist NECA. The structures represent an intermediate conformation between the inactive and active states, because they share all the features of GPCRs that are in a fully activated state, except that the cytoplasmic end of transmembrane helix 6 partially occludes the G protein binding site. The adenine substituent of the agonists bind in a similar fashion to the chemically-related region of the inverse agonist ZM241385⁸. Both agonists contain a ribose group, not found in ZM241385, which extends deep into the ligand binding pocket where it makes polar interactions with conserved residues in H7 (Ser277^{7 42} and His278⁷⁴³; superscripts refer to Ballesteros-Weinstein numbering ) and non-polar interactions with residues in H3. In contrast, the inverse agonist ZM241385 does not interact with any of these residues and comparison with the agonist-bound structures suggests that ZM241385 sterically prevents the conformational change in H5 and therefore it acts as an inverse agonist. Comparison of the agonist-bound structures of A^R with the agonist-bound structures of b-adrenoceptors suggests that the contraction of the ligand binding pocket caused by the inward motion of helices 3, 5 and 7 may be a common feature in the activation of all GPCRs. In the simplest model for the conformational dynamics of GPCRs¹⁰ there is an equilibrium between two states, R and R*. The inactive state R preferentially binds inverse agonists and the activated state R* preferentially binds agonists¹¹. Only R* can couple and activate G proteins. Although there are far more complex schemes¹² describing intermediates between R and R*, studies on rhodopsin have indicated that there is only one major conformational change that significantly alters the structure of the receptor³. Thus the structures of dark- state rhodopsin^13,14 and of opsin^15,16 are considered to be representative structures for the R and R* state, respectively. Structures of 6 different GPCRs^8,13,17"21 in conformations closely approximating to the R state have now been determined and it is clear that they are similar to each other, with RMSDs between any pair of structures in the transmembrane domains being less than 3 A. As observed in light-activation of rhodopsin, the major structural difference between R and R* is the movement of the cytoplasmic ends of helices 5 and 6 away from the receptor core by 5-6 A, opening up a cleft in the centre of the helix bundle where the C-terminus of a G protein can bind¹⁶. Recently, the structure of an agonist-bound beta-adrenoceptor (beta₂AR) was determined in complex with an antibody fragment (nanobody Nb80)⁵. This structure of beta₂AR is very similar to the structure of opsin, which suggests that the nanobody mimicked the action of a G protein by maintaining the receptor structure in an activated state. Given the structural similarities between opsin and the beta₂AR-Nb80 complex, it is likely that the structures of the R^* states of other GPCRs are also highly similar. This is consistent with the same heterotrimeric G proteins being able to couple to multiple different receptors²². However, do the conserved structures of R and R* imply that all agonists activate the receptors in an identical fashion? The recent structures of a thermostabilised beta^R bound to 4 different agonists suggested that a defining feature of agonist binding to this receptor is the formation of a hydrogen bond with Ser⁵⁴⁶ on transmembrane helix 5 that accompanies the contraction of the ligand binding pocket⁷. Here we describe two structures of the adenosine A2A receptor (A_2AR) bound to two different agonists, which suggests that the initial action of agonist binding to A^R has both similarities and differences compared to agonist binding in betaARs.

The native human A_¾ when bound to its endogenous agonist adenosine or to the high- affinity synthetic agonist NECA is unstable in detergent, so crystallization and structure determination relied on using a thermostabilised construct (A2AR-GL31 ) that contained four point mutations, which dramatically improved its thermostability. Pharmacological analysis showed that the mutant receptor bound the five antagonists tested with greatly reduced affinity (1.8 - 4.3 log units), whereas four agonists bound with similar affinity to the wild-type receptor (Fig. 5). However, A2AR-GL31 is only weakly activated by the agonist CGS21680 (Fig. 6), which suggests that the thermostabilising mutations might also decouple high-affinity agonist binding from the formation of R*. The conformation of GL31 is not consistent with it being in the fully-activated G protein coupled state, because we do not observe a 42-fold increase in affinity for NECA binding measured for G_as-coupled A^ ²³. These data all suggest that Aa_,R-GL31 is in an intermediate conformation between R and R*, which is consistent with the structural analysis presented below.

The two structures we have determined are of A2AR-GL31 bound to adenosine and NECA with resolutions of 3.0 A and 2.6 A, respectively (Table 1 ). Global alignments of the A^R- GL31 structures with A2A-T4L (A^R with T4 lysozyme inserted into inner loop 3) bound to the inverse agonist ZM241385 were performed based on those residues in the region of the ligand binding pocket that show the closest structural homology (Fig. 1 and "Structural changes in the thermostabilised A_2AR-GL31 with the bound agonist NECA", below/ This gave an rmsd in Calpha positions of 0.66 A for the 96 atoms selected, which include all residues involved in binding either adenosine or NECA, with the exception of those in H3. Using this transformation, the adenine-like moiety of the two ligands superimposes almost exactly (rmsd 0.56 A). The most significant differences between the two structures are seen in a distortion and a 2 A shift primarily along the helical axis of H3, a bulge in H5 (resulting from non-helical backbone conformation angles of residues Cys185 and Val186) that shifts residues into the binding pocket by up to 2 A and also a change in conformation of the cytoplasmic ends of H5, H6 and H7 (Fig. 1). Comparison of the A2AR-GL31 structure with the agonist-bound beta₂AR-Nb80 complex indicates that these differences are similar to the conformational changes in the beta₂AR that are proposed to be responsible for the formation of the R* state⁵. However, it is unlikely that the structure of A2AR-GL31 represents the fully activated state, because comparison with opsin bound to the C-terminal peptide of the G protein transducin shows that there is insufficient space in A2AR-GL31 for the C-terminus of the G protein to bind (Fig. 11). This is based on the assumption that all G proteins bind and activate GPCRs in a similar fashion, but given the highly-conserved structures of both G proteins and GPCRs this seems a reasonable hypothesis. The fact that the structure of A2_AR-GL31 represents an agonist-binding state is consistent with how A2A -GL31 was engineered. Thermostabilising mutations were selected by heating the NECA-bound detergent-solubilised receptor, so the mutations are anticipated to stabilize the agonist-bound state either by stabilizing helix-helix interactions and/or biasing the conformational equilibrium between the agonist-bound R* state and the agonist bound R- state^24"26. The two most thermostabilising mutations, L48A and Q89A, are in regions of the receptor that are involved in transitions between R and R*, providing a possible explanation for their thermostabilising effect (Fig. 8). The other two mutations, A54L and T65A, are at the receptor-lipid interface and the reason for their thermostabilising effect is unclear. Although the overall shape of the ligand binding pockets of A2_AR and beta₂AR are different, the structural similarities with the beta₂AR-Nb80⁵ and the structural differences to ZM241385- bound A2A-T4L⁸ indicate that the structure of the binding pocket in A2_AR-GL31 is a good representation of the agonist-bound binding pocket of the wild-type receptor (Fig.1).

Adenosine and NECA bind to A2_AR-GL31 in a virtually identical fashion, in addition, the adenine ring in the agonists interacts with A^R in a similar way to the chemically-related triazolotriazine ring of the inverse agonist ZM241385 (Fig. 2). Thus the hydrogen bonds between exocyclic adenosine N6 (Fig. 9) with both Glu169 in extracellular loop 2 (EL2) and Asn253⁶⁵⁵ in H6 are similar, with the significant ττ-stacking interaction with Phe168 in EL2 also conserved. One of the major structural differences between ZM241385 and the agonists is the presence of a furan substituent on C20 of triazolotriazine in the inverse agonist, whilst agonists contain a ribose substituent linked to N9 of adenine (Fig. 2). In ZM241385, the furan group forms a hydrogen bond with Asn253^{6 55} in H6 and van der Waals contacts with other residues in H3, H5 and H6⁸. In contrast, the ribose moiety in agonists forms hydrogen bonds with Ser277⁷⁴² and His278⁷⁴³ in H7, in addition to van der Waals interactions with other residues in H3 and H6 (Fig. 2). In particular, Val84³³² has to shift its position upon agonist binding due to a steric clash with the ribose ring, which may contribute to the 2 A shift observed in H3 (Fig. 3). These differences in binding between ZM241385 and either adenosine or NECA suggest that the residues that bind uniquely to agonists (Ser277⁷⁴² and His278⁷⁴³) play a key role in the activation of the receptor, as previously shown by mutagenesis studies^27,28. This is analogous to the situation in the activation of beta^R, where only full agonists cause the rotamer conformation changes of Ser⁵⁴⁵ in H5, whereas the inverse agonist ICI118551 prevents receptor activation by sterically blocking the rotamer change^7,29. However, the details of the activation differ in that the critical residues that bind agonists and not antagonists are in H5 in betaiAR, but in H7 in A_2AR (Fig. 4). Adenosine and NECA activate the A_2AR through interactions with H3 and H7 that are absent in the interactions between the receptor and the inverse agonist ZM241385 (Fig. 2). The inward shift of H7, the movement of H3 and the consequent formation of a bulge in H5 are all observed in the structure of agonist-bound A_2AR-GL31 and beta₂AR-Nb80 (Fig.1). The formation of the bulge in H5 of the beta₂AR-Nb80 structure was linked to a series of conformational changes that generates the 60^° rotation of H6 about Phe282⁶⁴⁴, resulting in the cytoplasmic end of H6 moving out from the receptor centre and opening the cleft where the C terminus of a G protein is predicted to bind^5,6. There are analogous side chain movements in A_2AR-GL31 that result in a 40^° rotation of H6, but the cytoplasmic end of H6 remains partially occluding the G protein-binding cleft (Fig. 7), perhaps because the fully active conformation requires the binding of G proteins to stabilize it. Interestingly, the structure of beta₂AR⁶ with a covalently bound agonist is also not in the fully activated R^* conformation, which is only seen after the nanobody Nb80 is bound⁵. The importance of the bulge in H5 in the activation of A_2AR is highlighted by how inverse agonists bind. Formation of the H5 bulge results in the inward movement of Cys185⁵⁴⁶ (Calpha moves by 4 A), which in turn causes the movement of Val186 and ultimately a shift of His250⁶⁵² by 2 A into the ligand binding pocket thereby sterically blocking the binding of ZM241385 (Fig. 10). Hence, when the inverse agonist binds, it is anticipated that the H5 bulge is unlikely to form due to the opposite series of events and hence the formation of the R* state is inhibited. Thus in both betaARs and A₂AR, the formation of the H5 bulge seems to be a common action of agonists, whereas inverse agonists seem to prevent its formation. However, the energetic contributions to its formation may be different between the two receptors. In the betaARs there is a major contribution from direct interaction between the agonist and Ser⁵⁴⁶, while in the A₂AR, the major interaction appears to come from steric interactions between the agonist and H3, combined with polar interactions involving residues in H7. Despite these differences, agonist binding to both receptors involves strong attractive non-covalent interactions that pull the extracellular ends of H3, H5 and H7 together.

Recently, a related manuscript appeared³⁰, describing the structure of the A₂A-T4L chimera bound to the agonist UK432097, which is identical to NECA except for two large substituents on the adenine ring. The structure of UK432097-bound A_2A-T4L is very similar to the structures presented here in the transmembrane regions (rmsd 0.6 A), although there are differences in the extracellular surface due to the bulky extensions of UK432097 interacting with the extracellular loops and the absence of density for residues 149-157. Xu ef a/, conclude that the structure of UK432097-bound A2A-T4L is in an "active state configuration", whereas we conclude that the NECA- and adenosine-bound structures are best defined as representing an intermediate state between R and R*.

Structural changes in the thermostabilised A_2AR-GL31 with the bound agonist NECA To facilitate a structural comparison between A^R with the inverse agonist ZM241385 bound and the thermostabilised A2A -GL31 with bound agonist, the structures were superimposed based on those residues in the region of the ligand binding pocket that show the closest structural homology. This was achieved using the lsq_improve option of the program O and an initial transformation based on residues at the C-terminus of H6 and the N-terminus of H7. The final superposition was based on residues 16-21 in H1 , 51-70 in H2 and ECL1 , 132-140 in H4 and ECL2, 142-146 in ECL2, 166-182 in ECL2 and H5 and 245-283 in H6, ECL3 and H7 and gave an rmsd in Ca positions of 0.66 A for the 96 atoms. This selection includes all residues involved in binding either ligand with the exception of those in H3. Using this transformation, the adenine-like moiety of ZM241385 and the adenine ring of NECA superimpose with an rmsd of 0.56 A (all atoms). Based on this superposition, there are significant shifts in several of the transmembrane helices with the largest in H3, 5, 6 and 7. In the following, the changes will be described in terms of the transition from the antagonist bound to the agonist bound structures. H1 undergoes a rotation of 7° about an axis approximately normal to the helix axis, resulting in shifts of ~2 A at either end, with the N- terminus moving inward towards the core of the helical bundle. H2 undergoes a similar, but slightly smaller, rotation, so that the side chain packing interactions between the two helices are well preserved. Superimposing the helices individually gives an rmsd of 0.37 A for H1 and 0.35 A for H2, demonstrating that the helices move as almost rigid groups. In particular, the mutations at residues Leu48, Ala54 and Thr65 have no significant effect on the conformation of H2. H3 displays a relatively large shift (~2 A) as well as a change in conformation with the distinct kink at residue Cys82 in the antagonist structure no longer present in the agonist bound form. The shift of H3 is both toward H6/H7 and upward along the helix axis. This results in changes in the interactions between H2 and H3, although the same residues are involved and there are no significant changes in side chain conformations. This change can be described as the hydrophobic interface of one helix sliding over that of the other. The N-terminal region of H4 follows the shift of H3, maintaining the helix-helix packing, but the C-terminal region shows only a small movement. H5 displays a significant local conformational change in the region of Cys185. While the N-terminal residues (174-182) superpose well, there is a shift of 3.8 A in the Ca of Cys185 towards H3. As a consequence, the C-terminal residues of H5 are shifted both outwards from the face of the receptor as defined by H5, 6 and 7 and also laterally towards H6, with a maximum shift of ~3 A. The helical conformation of residues 186-204 in H5 is unchanged, but several side chains in this region adopt different rotamer conformations due to changes in the interaction of H5 and H6. In particular, Leu190, Tyr197 and Phe201 all change d by ~120°. H6 undergoes the largest shift when comparing the two structures. As observed for H5, the region of the helix involved in forming the ligand-binding pocket (residues 245-256) adopts the same conformation, but there is a deformation of the helix over residues 241-244. This results in the lower (N-terminal) region of H6 being rotated by ~40° about the helix axis and displaced outward from the helical core by a maximum of 5.6 A at Val229. The rotation of H6 and its movement relative to H5 results in a complete change in the side chain packing between the two helices. The upper (N-terminal) region of H7 also superimposes well (rmsd 0.3 A for residues 267-283) but a sharp kink at Val283 produces a shift of 2.7 A for the Ca of Asn284. A further perturbation in H7 is due to a change in Pro285 from a trans to cis conformation. The net result is an inward movement of H7 by ~2 A and a remodeling of the H6-H7 interface in this region. Additional changes in helical conformation towards the C- terminus of H7 largely reverse the inward shift and produce an outward shift of ~1.5 A in H8 parallel to the helix axis.

Methods

Expression, purification and crystallization

The thermostabilised A2A -GL31 construct contains amino acid residues 1-316 of the human AZAR, four thermostabilising point mutations (L48A²⁴⁶, A54L^{2 52}, T65A^{2 63} and Q89A^{3 37}) and the mutation N154A to remove a potential N-glycosylation site. A2AR-GL31 was expressed in insect cells using the baculovirus expression system and purified in the detergent octylthioglucoside using Ni²⁺-NTA affinity chromatography and size exclusion chromatography (see Methods). The purified receptor was crystallized in the presence of cholesteryl hemisuccinate by vapour diffusion, with the conditions described below. Data collection, structure solution and refinement

Diffraction data were collected in multiple wedges (20^° per wedge) from a single cryo-cooled crystal (100 K) for the GL31-NECA complex at beamline ID23-2 at ESRF, Grenoble, France and from 4 crystals for the GL31 -adenosine complex, at beamline I24 at Diamond, Harwell, UK. The structures were solved by molecular replacement using the ZM241385-bound A^- T4L structure (PDB code 3EML)⁸ as a model (see Methods). Data collection and refinement statistics are presented in Table 1 and omit densities for the ligands are shown in Fig. 10.

Expression, purification and crystallization

The human A_¾ construct, GL31 , contains four thermostabilising point mutations (L48A²⁴⁶, A54L^{2 52}, T65A^{2 63} and Q89A^{3 37}), the mutation N154A to remove the potential N-glycosylation site and a truncation at the C-terminus after Ala316 (Ref ³²). A polyhistidine tag (His₁₀) was engineered at the C-terminus, separated from the receptor by a TEV protease cleavage site. Baculovirus expression and membrane preparation were performed as described previously for the betaiAR³³.

Membranes were thawed at room temperature, diluted with 25 mM Hepes pH 7.4, in presence of protease inhibitors (Complete™, Boehringer). Membranes were pre-incubated with NECA at 100 μΜ for 45 minutes at room temperature. The receptor-ligand complexes were then solubilised by adding decylmaltoside (DM) and NaCI to give final concentrations of 1.5% and 0.3M, respectively, stirred for 30 minutes (4^°C) and insoluble material removed by ultracentrifugation (120,000 g, 45 minutes, 4^°C). All protein purification steps were performed at 4°C. The solubilised receptor sample was filtered through a 0.22 μΐη filter (Millipore) and applied to a 5 ml Ni-NTA superflow cartridge (Qiagen) pre-equilibrated with buffer (25 mM Hepes, pH 7.4, 0.1 M NaCI, 100 μΜ NECA, 0.15% DM, 2.5 mM imidazole). The column was washed sequentially with the same buffer supplemented with either 10, 40 or 80 mM imidazole, and then eluted with 250 mM imidazole. The eluted receptor-ligand complex was mixed with His₆-tagged TEV protease to cleave the tag for 4-6 hours, 4^°C, concentrated to 2 ml using an Amicon-ultra spin concentrator (Ultracel-50K, Millipore) and then desalted using a PD-10 column (GE Healthcare). Eluted fractions were further purified by binding the TEV and other contaminants to Ni-NTA (QIAGEN) pre-equilibrated in 25 mM Hepes pH 7.4, 0.1 M NaCI, 100 μΜ NECA, 0.15% DM, 40 mM imidazole, incubating for 30 minutes and then collecting the flow-through. For detergent exchange into 0.35% octylthioglucoside (OTG), the sample was concentrated using an Amicon-ultra concentrator (Ultracel-50K, Millipore), diluted 10-fold in 25 mM Hepes pH 7.4, 0.1 M NaCI, 100 μΜ NECA, 0.35% OTG, and concentrated again to 0.3 ml. The protein sample was applied to a Superdex 200 10/300 GL size exclusion column pre-equilibrated in 25 mM Hepes pH 7.4, 0.1 M NaCI, 100 μ NECA, 0.35% OTG and run at 0.5 ml/minute. Eluted receptor fractions (2-2.5 ml) were concentrated to 50-60 μΙ. Protein determination was performed using the amido black³⁴ assay. Before crystallization, cholesteryl hemisuccinate (CHS) and OTG were added to 1 mg/ml and 0.5% respectively and the protein concentration adjusted to 10-12.5 mg/ml. NECA and adenosine A2A-GL31 crystal hits were obtained using a new PEG-based crystallisation screen developed in house³⁵. Crystals were grown at 4°C in 100 nl sitting drops using 0.05 M ADA NaOH, pH 6.4, 23.6% PEG 400, 4% v/v 2-propanol for the NECA complex. Crystals were cryo-protected by soaking in 0.05 M ADA NaOH, pH 6.4, 45% PEG 400. For the adenosine complex, crystals were initially grown in 0.05 M TrisHCI, pH 7.6, 9.6% PEG 200, 22.9%. PEG 300. Crystals were cryo-protected by soaking in 0.05 M TrisHCI, pH 7.5, 15% PEG 200, 30% PEG 300. The crystals were mounted on Hampton CrystalCap HT loops and cryo-cooled in liquid nitrogen.

Data collection, structure solution and refinement

Diffraction data for the NECA complex were collected at the European Synchrotron Radiation Facility, Grenoble with a Mar 225 CCD detector on the microfocus beamline ID23-2 (wavelength, 0.8726 A) using a 10 μηι focused beam and for the adenosine complex on beamline I24 at the Diamond Light Source, Harwell with a Pilatus 6M detector and a 10 μηη microfocus beam (wavelength 0.9778 A). The microfocus beam was essential for the location of the best diffracting parts of single crystals, as well as allowing several wedges to be collected from different positions. Images were processed with MOSFLM³⁶ and SCALA³⁷. The NECA complex was solved by molecular replacement with PHASER³⁸ using the A2A-T4L structure (PDB code 3EML)⁸ as a model after removal of the coordinates for T4L, all solvent molecules and the inverse agonist ZM241384. This structure was then used as a starting model for the structure solution of the adenosine complex. Refinement and rebuilding were carried out with REFMAC5³⁹ and COOT⁴⁰ respectively. Smile strings for NECA and adenosine were created using Sketcher and dictionary entries using Libcheck. Hydrogen bond assignments for the ligands were determined using HBPLUS⁴¹.

To facilitate a structural comparison between ZM241385-bound A_2A-T4L and the thermostabilised A_2A-GL31 with bound agonist, the structures were superimposed based on those residues in the region of the ligand binding pocket that show the closest structural homology. This was achieved using the Isqjmprove option of the program O⁴² and an initial transformation based on residues at the C-terminus of helix 6 and the N-terminus of helix 7. The final superposition, based on residues 16-21 in H1 , 51-70 in H2 and ECL1 , 132-140 in H4 and ECL2, 142-146 in ECL2, 166-182 in ECL2 and H5 and 245-283 in H6, ECL3 and H7, gave an rmsd in Ca positions of 0.66 A for the 96 atoms and includes almost all residues involved in binding either ligand with the exception of those in H3. Using this transformation, the adenine moiety of the agonist superimposes well with the equivalent atoms of the triazolotriazene bicyclic ring of ZM241385 (rmsd 0.56 A). Validation of the final refined models was carried out using Molprobity⁴³. Omit densities for the ligands are shown in Fig. 10. All figures in the manuscript were generated using either Pymol (DeLano Scientific) or CCPmg³¹.

Binding of agonists and antagonist to / ?-GZ.37 expressed in CHO cells

Chinese hamster ovary (CHO) cells were maintained in culture in DMEM HA s F12 media containing 10 % FBS. Cells were transfected with plasmids expressing either wild-type adenosine A^R or A2AR-GL31 using GeneJuice according to manufacturer's instructions (EMD Biosciences). 48h after transfection, cells were harvested, centrifuged at 200 g for 5 minutes at 4° C and the pellet re-suspended in 20 mM HEPES, 10 mM EDTA buffer (pH 7.4). The membrane suspension was homogenised and centrifuged at 200 g for 15 minutes at 4°C. The supernatant was collected, the pellet re-suspended in 20 mM HEPES, 10 mM EDTA (pH 7.4) buffer and the solution homogenised and centrifuged as described previously⁴⁴. The collected supernatant was centrifuged for 30 min at 40000 g at 4°C. Pellets were re-suspended in 20 mM Hepes, 0.1 mM EDTA to a protein concentration of 1 mg/ml and stored at -80°C .

Membranes from CHO cells transiently expressing wild-type or A2A -GL31 (10-15 μg/well) were assessed using competition [³H]NECA binding in buffer containing 50 mM Tris-HCI (pH 7.4) as described previously⁴⁴. Inhibition curves were fitted to a four-parameter logistic equation to determine IC₅₀ values which were converted to K| values using K_D values determined by saturation binding and the [³H]NECA concentration of 10 nM. G protein-coupling activity o†A_2AR-GL31 measured in whole cells

A2AR-His₆ and A2AR-GL31-H^'IS₆ (amino acid residues 1-316 of human A^R) were subcloned into plasmid pcDNA5/FRT/TO using Kpnl and Notl restriction sites. Flp-in T-Rex HEK293 cells were maintained at 37°C in a humidified atmosphere in Dulbecco's modified Eagle's medium without sodium pyruvate, supplemented with 4500 mg/L glucose, L-glutamine, 10% (v/v) FBS, 1 % penicillin/streptomycin mixture and 10 μg/mL blasticidin. To generate stable cell lines, the cells were transfected with a ratio of 1 :9 receptor cDNA in pcDNA5/FRT/TO vector and pOG44 vector using Genejuice as per manufacturer's instructions (EMD Biosciences). After 48 h, media was replaced with fresh medium supplemented with 200 μg/mL hygromycin B to select for stably expressing clones. Colonies were combined and tested for doxycycline-induced receptor expression. To induce receptor expression clones were treated with either 1 ng/mL or 3 ng/mL doxycyline for 16 h.

Cells were seeded at a density of 25,000 per well in a poly-L-lysine coated 96-well half area plate. Cells were induced with doxycyline (3 or 1 ng/mL) for 16 h. After 16 h media was removed and replaced with fresh media containing 100 μΜ Ro-201724 and 2 U/mL adenosine deaminase. Cells were incubated at 37°C for 30 min prior to addition of varying concentrations of agonist (25°C, 30 min). As a control cells were also incubated for 30 min (25°C) with 10 μΜ forskolin. Cells were then lysed and cAMP produced detected using the CisBio cAMP kit according to manufacturer's instructions before plates were read on a PolarStar fluorescence plate reader.

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58. Adams, P. D. et al. PHENIX: A comprehensive Python-based system for macromolecular structure solution. Acta Crystallogr D Biol Crystallogr 66, 213-221 , doi:S0907444909052925 [pii] 10.1 107/S0907444909052925 (2010). Example 2: RMSD calculations

The RMSDs were calculated as indicated: Both molecules were initially read into Maestro and their sequences were aligned using the 'Pairwise Alignment' algorithm contained within the 'Multiple Sequence Viewer' toolbar within Maestro. Manual adjustment within the 'Multiple Sequence Viewer' using the 'Grab and drag' tool was performed on the region between 149- 157, where the residues are not visible due to poor electron density, to ensure correct alignment of identically numbered residues. The identical residues in common between the two structures were then selected within the 'Multiple Sequence Viewer' using the 'Select and slide' tool, corresponding to residues 7-148, 158-208, 222-305 and excluding residues 54, 88, 107, 122, 202, 235, 239 and 277 (mutated for thermostabilisation). For the superposition the 'Superposition' tool was selected from the 'Tools' menu in the main window of Maestro. The 'Superimpose by ASL' tab was selected and the 'Select' button was pressed, in the 'Atom Selection' pop up box that appears 'Selection' was pressed to select the highlighted atoms. Additionally for the backbone RMSD, the 'Residue' tab was selected 'Backbone/side chain' was highlighted, the 'Backbone' checkbox was checked and 'Intersect' was pressed to select only the intersection between the backbone and the highlighted atoms. Finally 'OK' was selected. The RMSD is then returned in the box at the bottom of the 'Superposition' tool. Tables

Table (i) shows the x, y and z co-ordinates by amino acid residue of each non-hydrogen atom in the polypeptide structure for molecule NECA-bound A2A-GL31. The third column of the tables indicates whether the atom is from an amino acid residue of the protein (by e-letter amino acid code e.g. TRP, GLU, ALA etc), the NECA ligand (NEC), water (HOH) or octylthioglucoside, also known as 1-S-Octyl-beta-D-thiogJucopyranoside (SOG).

Table (i)

Parameters for modelling the C121 crystal system NECA-A_2AR-GL31 complex.

1 N ILE A 3 35. 343 -2. , 057 -30. 022

2 CA ILE A 3 36. 094 -0. ,818 -29. 646

3 CB ILE A 3 37. 602 -1. .118 -29. 355

4 CGI ILE A 3 37. 765 -1. ,870 -28. 032

5 CD1 ILE A 3 37. 807 -3. .368 -28. 171

6 CG2 ILE A 3 38. 420 0. , 146 -29. 259

7 C ILE A 3 35. 911 0. .303 -30. 683

8 O ILE A 3 36. 033 1. ,484 -30. 348

9 N MET A 4 35. 590 -0. , 074 -31. 922

10 CA MET A 4 35. 353 0. ,879 -33. 020

11 CB MET A 4 34. 631 0. ,186 -34. 187

12 CG MET A 4 34. 387 1. .081 -35. 404

13 SD MET A 4 33. 005 0. .535 -36. 431

14 CE MET A 4 32. 708 2. .005 -37. 414

15 C MET A 4 34. 579 2. .132 -32. 580

16 O MET A 4 35. 002 3. .257 -32. 858

17 N GLY A 5 33. 456 1. .925 -31. 893

18 CA GLY A 5 32. 622 3. .022 -31. 402

19 C GLY A 5 31. 273 3. .107 -32. 093

20 O GLY A 5 31. 035 2. .428 -33. 093

21 N SER A 6 30. 390 3. .945 -31. 552

22 CA SER A 6 29. 057 4. .148 -32. 116

23 CB SER A 6 27. 977 3. .736 -31. 109

24 OG SER A 6 26. 683 4. .114 -31. 549

25 C SER A 6 28. 863 5. .600 -32. 546

26 0 SER A 6 28. 961 6, .512 -31. 726

27 N SER A 7 28. 580 5. .795 -33. 834

28 CA SER A 7 28. 404 7. .126 -34. 428

29 CB SER A 7 28. 286 7. .021 -35. 952

30 OG SER A 7 29. 303 6. .195 -36. 489

31 C SER A 7 27. 181 7. .847 -33. 873

32 O SER A 7 27. 174 9. .077 -33. 759

33 N VAL A 8 26. 149 7. .072 -33. 540

34 CA VAL A 8 24. 928 7, .605 -32. 939

35 CB VAL A 8 23. 840 6. .511 -32. 762

36 CGI VAL A 8 22. 493 7. .144 -32. 422

37 CG2 VAL A 8 23. 720 5. .648 -34. 015

38 C VAL A 8 25. 252 8. .241 -31. 586

39 O VAL A 8 24. 803 9. .354 -31. 291

40 N TYR A 9 26. 042 7. .528 -30. 779

41 CA TYR A 9 26. 496 8. .035 -29. 487

42 CB TYR A 9 27. 300 6. .971 -28. 718

43 CG TYR A 9 28. 171 7. .550 -27. 623

44 CD1 TYR A 9 27. 611 8. .013 -26. 427

45 CE1 TYR A 9 28. 408 8. .559 -25. 424

46 CZ TYR A 9 29. 781 8. .648 -25. 612

47 OH TYR A 9 30. 574 9. .186 -24. 626

48 CE2 TYR A 9 30. 359 8. .196 -26. 790

49 CD2 TYR A 9 29. 552 7. .652 -27. 788

50 C TYR A 9 27. 306 9. .322 -29. 645 51 0 TYR A 9 27.,021 10., 318 -28.978

52 N ILE A 10 28. ,299 9. .291 -30. 534

53 CA ILE A 10 29. ,171 10. ,441 -30. 790

54 CB ILE A 10 30. , 156 10. .183 -31. 973

55 CGI ILE A 10 30. ,804 8. .788 -31. 893

56 CD1 ILE A 10 31. ,767 8. ,564 -30. 741

57 CG2 ILE A 10 31. ,199 11. .300 -32. 079

58 C ILE A 10 28. ,366 11. .697 -31. 105

59 0 ILE A 10 28. ,637 12. ,764 -30. 551

60 N THR A 11 27, ,375 11. ,561 -31. 987

61 CA THR A 11 26. ,623 12. .719 -32. 486

62 CB THR A 11 25. ,970 12. ,449 -33. 886

63 OG1 THR A 11 25. .413 13. , 663 -34. 403

64 CG2 THR A 11 24. ,883 11. .382 -33. 820

65 C THR A 11 25. , 632 13. .284 -31. 453

66 0 THR A 11 25. ,514 14. .508 -31. 307

67 N VAL A 12 24. , 947 12. .394 -30. 731

68 CA VAL A 12 24. .091 12. .792 -29. 605

69 CB VAL A 12 23. ,350 11. .577 -28. 973

70 CGI VAL A 12 22. ,704 11. .956 -27. 640

71 CG2 VAL A 12 22. ,298 11. .025 -29. 931

72 C VAL A 12 24. , 933 13. .510 -28. 548

73 0 VAL A 12 24. ,524 14. .540 -28. Oil

74 N GLU A 13 26. ,118 12. .965 -28. 280

75 CA GLU A 13 27. ,067 13. .553 -27. 340

76 CB GLU A 13 28. ,272 12. .619 -27. 153

77 CG GLU A 13 29. ,205 12. .966 -25. 993

78 CD GLU A 13 28. ,784 12. .369 -24. 651

79 OE1 GLU A 13 27. , 666 12. .657 -24. 173

80 OE2 GLU A 13 29. ,592 11. .625 -24. 057

81 C GLU A 13 27. ,511 14. .942 -27. 804

82 0 GLU A 13 27. , 635 15. .863 -26. 995

83 N LEU A 14 27. ,729 15. .088 -29. 110

84 CA LEU A 14 28. ,067 16. .383 -29. 708

85 CB LEU A 14 28. ,470 16. .214 -31. 175

86 CG LEU A 14 29. ,891 15. .718 -31. 454

87 CD1 LEU A 14 29. , 950 15. .009 -32. 802

88 CD2 LEU A 14 30. , 912 16. .857 -31. 380

89 C LEU A 14 26. ,929 17. .396 -29. 583

90 0 LEU A 14 27. , 170 18. .577 -29. 316

91 N ALA A 15 25. , 695 16. .929 -29. 769

92 CA ALA A 15 24. ,511 17. .781 -29. 621

93 CB ALA A 15 23. .243 16. .989 -29. 905

94 C ALA A 15 24. , 452 18. .416 -28. 231

95 0 ALA A 15 24. ,298 19. .637 -28. 102

96 N ILE A 16 24. , 602 17. .577 -27. 204

97 CA ILE A 16 24. ,570 18. .007 -25. 807

98 CB ILE A 16 24. .782 16. .816 -24. 852

99 CGI ILE A 16 23. , 635 15. .817 -24. 997

100 CD1 ILE A 16 23. ,965 14. ,449 -24. 485

101 CG2 ILE A 16 24. , 917 17. .293 -23. 388

102 C ILE A 16 25. , 613 19. .081 -25. 526

103 0 ILE A 16 25. ,315 20. .077 -24. 859

104 N ALA A 17 26. ,825 18. ,874 -26. 044

105 CA ALA A 17 27. , 911 19. ,848 -25. 911

106 CB ALA A 17 29. ,164 19. ,359 -26. 628

107 C ALA A 17 27. ,499 21. ,224 -26. 425

108 0 ALA A 17 27. ,745 22. ,235 -25. 764

109 N VAL A 18 26. ,854 21. ,252 -27. 592

110 CA VAL A 18 26. ,378 22. .504 -28. 179 111 CB VAL A 18 25,.849 22.315 -29.626

112 CGI VAL A 18 25. .273 23. 617 -30. 159

113 CG2 VAL A 18 26. .966 21. 827 -30. 545

114 C VAL A 18 25. .328 23. 156 -27. 276

115 0 VAL A 18 25. .491 24. 314 -26. 877

116 N LEU A 19 24. .279 22. 407 -26. 931

117 CA LEU A 19 23. ,245 22. 906 -26. 006

118 CB LEU A 19 22. .136 21. 871 -25. 806

119 CG LEU A 19 21. .213 21. 604 -27. 001

120 CD1 LEU A 19 20. .265 20. 463 -26. 687

121 CD2 LEU A 19 20. .433 22. 861 -27. 417

122 C LEU A 19 23. .795 23. 366 -24. 649

123 0 LEU A 19 23. .298 24. 333 -24. 075

124 N ALA A 20 24. .822 22. 678 -24. 148

125 CA ALA A 20 25. .492 23. 077 -22. 910

126 CB ALA A 20 26. .450 21. 989 -22. 439

127 C ALA A 20 26. .220 24. 416 -23. 054

128 0 ALA A 20 26. .115 25. 282 -22. 180

129 N ILE A 21 26. .947 24. 585 -24. 159

130 CA ILE A 21 27. .626 25. 853 -24. 448

131 CB ILE A 21 28. .645 25. 723 -25. 615

132 CGI ILE A 21 29. .734 24. 700 -25. 257

133 CD1 ILE A 21 30. .373 24. 010 -26. 456

134 CG2 ILE A 21 29. .282 27. 081 -25. 935

135 C ILE A 21 26, .625 26. 995 -24. 699

136 0 ILE A 21 26. .778 28. 082 -24. 139

137 N LEU A 22 25, .594 26. 734 -25. 505

138 CA LEU A 22 24. .589 27. 757 -25. 850

139 CB LEU A 22 23. .562 27. 218 -26. 857

140 CG LEU A 22 23. .989 26. 908 -28. 299

141 CD1 LEU A 22 22. .829 26. 284 -29. 065

142 CD2 LEU A 22 24. .521 28. 144 -29. 024

143 C LEU A 22 23. .862 28. 312 -24. 631

144 0 LEU A 22 23. .868 29. 527 -24. 399

145 N GLY A 23 23. .245 27. 418 -23. 857

146 CA GLY A 23 22. .485 27. 799 -22. 670

147 C GLY A 23 23. .287 28. 509 -21. 592

148 0 GLY A 23 22. .773 29. 396 -20. 908

149 N ASN A 24 24. .552 28. 133 -21. 442

150 CA ASN A 24 25. ,357 28. 657 -20. 346

151 CB ASN A 24 26. .238 27. 557 -19. 753

152 CG ASN A 24 25. ,426 26. 530 -18. 983

153 ODl ASN A 24 24. .932 26. 807 -17. 889

154 ND2 ASN A 24 25. ,263 25. 350 -19. 563

155 C ASN A 24 26. .136 29. 914 -20. 676

156 0 ASN A 24 26. ,371 30. 743 -19. 794

157 N VAL A 25 26. ,522 30. 064 -21. 944

158 CA VAL A 25 26. ,940 31. 370 -22. 461

159 CB VAL A 25 27. ,432 31. 294 -23. 930

160 CGI VAL A 25 27. ,437 32. 671 -24. 581

161 CG2 VAL A 25 28. ,822 30. 685 -23. 992

162 C VAL A 25 25. .766 32. 347 -22. 315

163 0 VAL A 25 25. , 941 33. 468 -21. 839

164 N LEU A 26 24. ,570 31. 894 -22. 693

165 CA LEU A 26 23. ,345 32. 671 -22. 515

166 CB LEU A 26 22. ,123 31. 847 -22. 938

167 CG LEU A 26 20. ,892 32. 569 -23. 501

168 CD1 LEU A 26 21. ,203 33. 265 -24. 831

169 CD2 LEU A 26 19. ,736 31. 593 -23. 675

170 C LEU A 26 23. ,191 33. 187 -21. 077 171 0 LEU A 26 22.940 34 ,.381 -20.869

172 N VAL A 27 23. 366 32, .294 -20. 098

173 CA VAL A 27 23. 299 32, .650 -18. 674

174 CB VAL A 27 23. 460 31, .399 -17. 750

175 CGI VAL A 27 23. 630 31, .799 -16. 284

176 CG2 VAL A 27 22. 277 30, .450 -17. 898

177 C VAL A 27 24. 340 33, .708 -18. 308

178 0 VAL A 27 24. 019 34 , .697 -17. 644

179 N CYS A 28 25. 579 33, .499 -18. 745

180 CA CYS A 28 26. 684 34 , .400 -18. 402

181 CB CYS A 28 28. 034 33, .761 -18. 732

182 SG CYS A 28 28. 469 32. .375 -17. 665

183 C CYS A 28 26. 563 35, .753 -19. 089

184 0 CYS A 28 26. 797 36, .791 -18. 463

185 N TRP A 29 26. 205 35, .726 -20. 373

186 CA TRP A 29 25. 942 36, .935 -21. 157

187 CB TRP A 29 25. 433 36, .561 -22. 554

188 CG TRP A 29 25. 833 37, .506 -23. 662

189 CD1 TRP A 29 26. 072 38, .850 -23. 559

190 NE1 TRP A 29 26. 401 39, .369 -24. 789

191 CE2 TRP A 29 26. 366 38, .364 -25. 721

192 CD2 TRP A 29 26. 006 37, .174 -25. 047

193 CE3 TRP A 29 25. 900 35. .984 -25. 783

194 CZ3 TRP A 29 26. 155 36, .020 -27. 155

195 CH2 TRP A 29 26. 511 37, .222 -27. 796

196 CZ2 TRP A 29 26. 622 38, .400 -27. 099

197 C TRP A 29 24. 902 37. .806 -20. 452

198 0 TRP A 29 25. 078 39. .022 -20. 328

199 N ALA A 30 23. 834 37. .164 -19. 976

200 CA ALA A 30 22. 741 37. .844 -19. 282

201 CB ALA A 30 21. 626 36. .859 -18. 951

202 C ALA A 30 23. 182 38. .601 -18. 028

203 0 ALA A 30 22. 637 39. .659 -17. 725

204 N VAL A 31 24. 164 38. .069 -17. 307

205 CA VAL A 31 24. 663 38. .729 -16. 091

206 CB VAL A 31 25. 413 37. .746 -15. 153

207 CGI VAL A 31 25. 799 38. .427 -13. 844

208 CG2 VAL A 31 24. 552 36. .526 -14. 866

209 C VAL A 31 25. 557 39. .926 -16. 420

210 0 VAL A 31 25. 584 40. .911 -15. 678

211 N TRP A 32 26. 277 39. .838 -17. 535

212 CA TRP A 32 27. 183 40. .906 -17. 950

213 CB TRP A 32 28. 171 40. .402 -19. 009

214 CG TRP A 32 29. 148 41. .445 -19. 487

215 CD1 TRP A 32 29. 919 42. .267 -18. 713

216 NE1 TRP A 32 30. 689 43. ,083 -19. 508

217 CE2 TRP A 32 30. 434 42. .793 -20. 823

218 CD2 TRP A 32 29. 467 41. ,762 -20. 850

219 CE3 TRP A 32 29. 028 41. .277 -22. 091

220 CZ3 TRP A 32 29. 563 41. .833 -23. 254

221 CH2 TRP A 32 30. 524 42. .859 -23. 193

222 CZ2 TRP A 32 30. 971 43. ,351 -21. 992

223 C TRP A 32 26. 420 42. , 140 -18. 439

224 0 TRP A 32 26. 762 43. ,266 -18. 075

225 N LEU A 33 25. 383 41. , 917 -19. 248

226 CA LEU A 33 24. 540 43. ,001 -19. 760

227 CB LEU A 33 23. 639 42. ,505 -20. 900

228 CG LEU A 33 24. 273 42. ,053 -22. 221

229 CD1 LEU A 33 23. 210 41. ,461 -23. 128

230 CD2 LEU A 33 25. 005 43. ,192 -22. 927 231 C LEU A 33 23.687 43..624 -18.655

232 0 LEU A 33 23. 873 44. .790 -18 .296

233 N ASN A 34 22. 762 42, .831 -18 .118

234 CA ASN A 34 21. 827 43, .284 -17 .094

235 CB ASN A 34 20. 733 42, .231 -16 .889

236 CG ASN A 34 19. 614 42, .708 -15 .982

237 ODl ASN A 34 19. 787 43, .627 -15 .181

238 ND2 ASN A 34 18. 454 42, .072 -16 .102

239 C ASN A 34 22. 518 43, .617 -15 .771

240 0 ASN A 34 23. 224 42, .788 -15 .200

241 N SER A 35 22. 293 44 , .837 -15 .294

242 CA SER A 35 22. 904 45, .328 -14 .062

243 CB SER A 35 22. 997 46, .856 -14 .096

244 OG SER A 35 23. 570 47 , .362 -12 .903

245 C SER A 35 22. 161 44 , .865 -12 .803

246 0 SER A 35 22. 735 44 , .854 -11 .711

247 N ASN A 36 20. 890 44 , .494 -12 .960

248 CA ASN A 36 20. 080 43, .951 -11 .862

249 CB ASN A 36 18. 597 43, .887 -12 .252

250 CG ASN A 36 18. 002 45, .254 -12 .551

251 ODl ASN A 36 17. 999 46, .148 -11 .703

252 ND2 ASN A 36 17. 478 45, .413 -13 .761

253 C ASN A 36 20. 548 42. .554 -11 .459

254 0 ASN A 36 20. 379 42, .132 -10 .310

255 N LEU A 37 21. 133 41. .847 -12 .421

256 CA LEU A 37 21. 619 40. .494 -12 .210

257 CB LEU A 37 21. 615 39. .720 -13 .537

258 CG LEU A 37 20. 255 39. .475 -14 .201

259 CD1 LEU A 37 20. 401 38. .688 -15 .493

260 CD2 LEU A 37 19. 304 38. .756 -13 .258

261 C LEU A 37 23. 009 40. .467 -11 .564

262 0 LEU A 37 23. 520 39. .398 -11 .229

263 N GLN A 38 23. 606 41. .642 -11 .374

264 CA GLN A 38 24. 973 41. .738 -10 .855

265 CB GLN A 38 25. 740 42. .877 -11 .544

266 CG GLN A 38 26. 147 42. .541 -12 .979

267 CD GLN A 38 26. 647 43. .734 -13 .775

268 OE1 GLN A 38 27. 127 44. .720 -13 .216

269 NE2 GLN A 38 26. 543 43. .641 -15 .098

270 C GLN A 38 25. 040 41. ,855 -9 .332

271 O GLN A 38 25. 518 42. ,856 -8 .795

272 N ASN A 39 24. 554 40. .823 -8 .646

273 CA ASN A 39 24. 686 40. ,718 -7 .192

274 CB ASN A 39 23. 348 40. , 995 -6 .489

275 CG ASN A 39 22. 326 39. .900 -6 .721

276 ODl ASN A 39 21. 821 39. ,727 -7 .832

277 ND2 ASN A 39 22. 012 39. , 155 -5 .667

278 C ASN A 39 25. 285 39. ,372 -6 .754

279 O ASN A 39 25. 387 38. ,439 -7 .557

280 N VAL A 40 25. 665 39. ,290 -5 .478

281 CA VAL A 40 26. 411 38. ,148 -4 .917

282 CB VAL A 40 26. 643 38. ,316 -3 .387

283 CGI VAL A 40 27. 349 37. ,098 -2 .801

284 CG2 VAL A 40 27. 446 39. 587 -3 .100

285 C VAL A 40 25. 788 36. 776 -5 .217

286 O VAL A 40 26. 491 35. 856 -5 .635

287 N THR A 41 24. 477 36. 657 -5 .014

288 CA THR A 41 23. 732 35. 416 -5 .264

289 CB THR A 41 22. 200 35. 656 -5 .121

290 OG1 THR A 41 21. 921 36. 163 -3 .810 291 CG2 THR A 41 21.393 34.370 -5.344

292 C THR A 41 24 .034 34 .790 -6 .632

293 0 THR A 41 24 .009 33 .567 -6 .780

294 N ASN A 42 24 .327 35 .632 -7 .619

295 CA ASN A 42 24 .519 35 .171 -8 .992

296 CB ASN A 42 23 .991 36 .206 -9 .982

297 CG ASN A 42 22 .482 36 .356 -9 .902

298 ODl ASN A 42 21 .739 35 .442 -10 .262

299 ND2 ASN A 42 22 .021 37 .505 -9 .413

300 C ASN A 42 25 .943 34 .734 -9 .325

301 0 ASN A 42 26 .173 34 .091 -10 .348

302 N TYR A 43 26 .886 35 .077 -8 .449

303 CA TYR A 43 28 .249 34 .540 -8 .502

304 CB TYR A 43 29 .080 35 .036 -7 .310

305 CG TYR A 43 29 .514 36 .492 -7 .378

306 CD1 TYR A 43 28 .670 37 .482 -7 .904

307 CE1 TYR A 43 29 .068 38 .819 -7 .960

308 CZ TYR A 43 30 .316 39 .181 -7 .474

309 OH TYR A 43 30 .712 40 .498 -7 .523

310 CE2 TYR A 43 31 .167 38 .223 -6 .938

311 CD2 TYR A 43 30 .760 36 .887 -6 .889

312 C TYR A 43 28 .217 33 .009 -8 .516

313 0 TYR A 43 28 .984 32 .377 -9 .245

314 N PHE A 44 27 .316 32 .425 -7 .723

315 CA PHE A 44 27 .145 30 .977 -7 .677

316 CB PHE A 44 26 .321 30 .563 -6 .455

317 CG PHE A 44 26 .906 31 .017 -5 .141

318 CD1 PHE A 44 26 .265 31 .994 -4 .381

319 CE1 PHE A 44 26 .801 32 .426 -3 .159

320 CZ PHE A 44 27 .989 31 .869 -2 .689

321 CE2 PHE A 44 28 .641 30 .890 -3 .446

322 CD2 PHE A 44 28 .097 30 .472 -4 .664

323 C PHE A 44 26 .527 30 .423 -8 .962

324 0 PHE A 44 26 .854 29 .312 -9 .388

325 N VAL A 45 25 .646 31 .204 -9 .581

326 CA VAL A 45 25 .031 30 .826 -10 .859

327 CB VAL A 45 23 .732 31 .644 -11 .136

328 CGI VAL A 45 23 .138 31 .311 -12 .497

329 CG2 VAL A 45 22 .698 31 .387 -10 .040

330 C VAL A 45 26 .036 30 .946 -12 .012

331 0 VAL A 45 26 .071 30 .087 -12 .904

332 N VAL A 46 26 .860 31 .996 -11 .978

333 CA VAL A 46 27 .890 32 .218 -13 .003

334 CB VAL A 46 28 .535 33 .633 -12 .904

335 CGI VAL A 46 29 .676 33 .794 -13 .922

336 CG2 VAL A 46 27 .489 34 .723 -13 .131

337 C VAL A 46 28 .965 31 .129 -12 .959

338 0 VAL A 46 29 .380 30 .624 -14 .008

339 N SER A 47 29 .393 30 .765 -11 .748

340 CA SER A 47 30 .351 29 .675 -11 .544

341 CB SER A 47 30 .590 29 .419 -10 .054

342 OG SER A 47 31 .004 30, .593 -9 .383

343 C SER A 47 29 .855 28, .391 -12 .184

344 0 SER A 47 30 .573 27, .763 -12 .970

345 N ALA A 48 28 .618 28, .022 -11 .849

346 CA ALA A 48 28 .012 26, .792 -12 .338

347 CB ALA A 48 26, .651 26, .582 -11 .700

348 C ALA A 48 27 , .912 26, .797 -13 .857

349 0 ALA A 48 28, .246 25, .805 -14 .517

350 N ALA A 49 27, .473 27, .928 -14 .402 351 CA ALA A 49 27.362 28..104 -15.846

352 CB ALA A 49 26. 647 29. .414 -16 .177

353 C ALA A 49 28. 730 28. .030 -16 .529

354 0 ALA A 49 28. 856 27. .415 -17 .590

355 N ALA A 50 29. 745 28. .644 -15 .915

356 CA ALA A 50 31. 128 28. .561 -16 .418

357 CB ALA A 50 32. 066 29. .401 -15 .560

358 C ALA A 50 31. 627 27. .113 -16 .507

359 0 ALA A 50 32. 117 26. .682 -17 .555

360 N ALA A 51 31. 473 26. .369 -15 .409

361 CA ALA A 51 31. 838 24. .949 -15 .337

362 CB ALA A 51 31. 585 24. .407 -13 .932

363 C ALA A 51 31. 131 24. .084 -16 .382

364 0 ALA A 51 31. 740 23. .174 -16 .949

365 N ASP A 52 29. 855 24. .369 -16 .639

366 CA ASP A 52 29. 087 23. .641 -17 .655

367 CB ASP A 52 27. 603 23. .993 -17 .576

368 CG ASP A 52 26. 924 23. .425 -16 .341

369 OD1 ASP A 52 27. 271 22. .305 -15 .897

370 OD2 ASP A 52 26. 022 24. .108 -15 .819

371 C ASP A 52 29. 602 23. .902 -19 .071

372 0 ASP A 52 29. 552 23. .016 -19 .933

373 N ILE A 53 30. 084 25. .126 -19 .301

374 CA ILE A 53 30. 733 25. .476 -20 .560

375 CB ILE A 53 31. 099 26. .983 -20 .634

376 CGI ILE A 53 29. 827 27. .835 -20 .752

377 CD1 ILE A 53 29. 972 29. .252 -20 .239

378 CG2 ILE A 53 32. 029 27. .257 -21 .819

379 C ILE A 53 31. 961 24. .588 -20 .748

380 0 ILE A 53 32. 080 23. .909 -21 .769

381 N LEU A 54 32. 844 24. .561 -19 .749

382 CA LEU A 54 34. 031 23. .698 -19 .802

383 CB LEU A 54 34. 911 23. .871 -18 .556

384 CG LEU A 54 35. 622 25. .217 -18 .326

385 CD1 LEU A 54 36. 421 25. .209 -17 .021

386 CD2 LEU A 54 36. 535 25. .592 -19 .496

387 C LEU A 54 33. 685 22. .219 -20 .028

388 0 LEU A 54 34. 464 21. .492 -20 .642

389 N VAL A 55 32. 521 21. .785 -19 .544

390 CA VAL A 55 32. 038 20. .424 -19 .800

391 CB VAL A 55 30. 786 20. .075 -18 .931

392 CGI VAL A 55 30. 107 18. .787 -19 .408

393 CG2 VAL A 55 31. 161 19. .963 -17 .455

394 C VAL A 55 31. 733 20. ,221 -21 .287

395 0 VAL A 55 32. 028 19. ,161 -21 .856

396 N GLY A 56 31. 143 21. .247 -21 .904

397 CA GLY A 56 30. 795 21. ,231 -23 .324

398 C GLY A 56 32. 013 21. ,371 -24 .215

399 0 GLY A 56 32. 148 20. , 656 -25 .209

400 N VAL A 57 32. 902 22. ,292 -23 .856

401 CA VAL A 57 34. 099 22. ,558 -24 .649

402 CB VAL A 57 34. 711 23. , 943 -24 .322

403 CGI VAL A 57 35. 919 24. ,222 -25, .212

404 CG2 VAL A 57 33. 673 25. ,040 -24, .491

405 C VAL A 57 35. 161 21. ,462 -24 , .481

406 0 VAL A 57 35. 655 20. , 917 -25, .473

407 N LEU A 58 35. 503 21. ,143 -23, .232

408 CA LEU A 58 36. 620 20. 236 -22, .939

409 CB LEU A 58 37. 517 20. 821 -21, .839

410 CG LEU A 58 38. 169 22. 192 -22, .048 411 CDl LEU A 58 38..647 22.755 -20.722

412 CD2 LEU A 58 39. .322 22 .134 -23. 048

413 C LEU A 58 36. .197 18 .808 -22. 576

414 0 LEU A 58 36. .690 17 .853 -23. 173

415 N ALA A 59 35. .280 18 .668 -21. 616

416 CA ALA A 59 34. .959 17 .358 -21. 036

417 CB ALA A 59 34. .139 17 .505 -19. 757

418 C ALA A 59 34. .290 16 .385 -21. 996

419 0 ALA A 59 34. .684 15 .219 -22. 058

420 N ILE A 60 33. .285 16 .851 -22. 735

421 CA ILE A 60 32, .582 15 .989 -23. 697

422 CB ILE A 60 31. .231 16 .595 -24. 174

423 CGI ILE A 60 30. .219 16 .619 -23. 020

424 CDl ILE A 60 28. .860 17 .250 -23. 360

425 CG2 ILE A 60 30, .674 15 .807 -25. 355

426 C ILE A 60 33. .463 15 .544 -24. 885

427 0 ILE A 60 33. .445 14 .363 -25. 250

428 N PRO A 61 34. .240 16 .477 -25. 486

429 CA PRO A 61 35. .176 16 .034 -26. 531

430 CB PRO A 61 35. .922 17 .321 -26. 901

431 CG PRO A 61 34. .952 18 .393 -26. 613

432 CD PRO A 61 34, .221 17 .948 -25. 374

433 C PRO A 61 36. .154 14 .968 -26. 028

434 0 PRO A 61 36. .380 13 .972 -26. 720

435 N PHE A 62 36. .702 15 .170 -24. 828

436 CA PHE A 62 37. .559 14 .172 -24. 178

437 CB PHE A 62 38. .104 14 .695 -22. 847

438 CG PHE A 62 39. .121 15 .796 -22. 980

439 CDl PHE A 62 39. .758 16 .050 -24. 191

440 CE1 PHE A 62 40. .699 17 .068 -24. 302

441 CZ PHE A 62 41. .034 17 .832 -23. 193

442 CE2 PHE A 62 40. .417 17 .581 -21. 974

443 CD2 PHE A 62 39. .468 16 .562 -21. 872

444 C PHE A 62 36. .833 12 .844 -23. 940

445 0 PHE A 62 37. .422 11 .775 -24. 101

446 N ALA A 63 35. .558 12 .920 -23. 565

447 CA ALA A 63 34. .757 11 .730 -23. 306

448 CB ALA A 63 33. .432 12 .105 -22. 647

449 C ALA A 63 34. .517 10 .932 -24. 582

450 0 ALA A 63 34. ,509 9 .704 -24. 550

451 N ILE A 64 34. .321 11 .637 -25. 697

452 CA ILE A 64 34. .150 11 .001 -27. 009

453 CB ILE A 64 33. .822 12 .037 -28. 127

454 CGI ILE A 64 32. .385 12 .550 -27. 976

455 CDl ILE A 64 32. .015 13 .686 -28. 917

456 CG2 ILE A 64 34. .002 11 .420 -29. 519

457 C ILE A 64 35. .388 10 .179 -27. 379

458 0 ILE A 64 35. ,276 9 .020 -27. 788

459 N ALA A 65 36. .560 10 .789 -27. 218

460 CA ALA A 65 37. ,834 10 .134 -27. 505

461 CB ALA A 65 38. ,991 11 .084 -27. 201

462 C ALA A 65 37. ,989 8 .820 -26. 735

463 0 ALA A 65 38. , 302 7 .781 -27. 322

464 N ILE A 66 37. , 732 8 .873 -25. 427

465 CA ILE A 66 37. ,874 7 .717 -24. 534

466 CB ILE A 66 37. , 673 8 .122 -23. 040

467 CGI ILE A 66 38. , 635 9 .245 -22. 627

468 CDl ILE A 66 40. ,104 8 .986 -22. 928

469 CG2 ILE A 66 37. ,850 6 .936 -22. 115

470 C ILE A 66 36. , 944 6 .552 -24. 899 471 0 ILE A 66 37.188 5.413 -24.496

472 N SER A 67 35 .892 6. 833 -25. 667

473 CA SER A 67 34 .946 5. 793 -26. 085

474 CB SER A 67 33 .558 6. 385 -26. 345

475 OG SER A 67 33 .552 7. 202 -27. 502

476 C SER A 67 35 .429 5. 003 -27. 306

477 0 SER A 67 34 .902 3. 927 -27. 599

478 N THR A 68 36 .441 5. 535 -27. 993

479 CA THR A 68 36 .916 4. 969 -29. 260

480 CB THR A 68 37 .475 6. 053 -30. 213

481 OG1 THR A 68 38 .690 6. 593 -29. 676

482 CG2 THR A 68 36 .455 7. 171 -30. 442

483 C THR A 68 37 .993 3. 909 -29. 087

484 0 THR A 68 38 .272 3. 152 -30. 014

485 N GLY A 69 38 .608 3. 872 -27. 910

486 CA GLY A 69 39 .691 2. 931 -27. 636

487 C GLY A 69 40 .856 3. 099 -28. 590

488 0 GLY A 69 41 .382 2. 117 -29. 117

489 N PHE A 70 41 .246 4. 352 -28. 815

490 CA PHE A 70 42 .388 4. 675 -29. 664

491 CB PHE A 70 42 .385 6. 167 -30. 035

492 CG PHE A 70 42 .535 7. 095 -28. 860

493 CD1 PHE A 70 41 .422 7. 500 -28. 128

494 CE1 PHE A 70 41 .553 8. 363 -27. 041

495 cz PHE A 70 42 .809 8. 833 -26. 682

496 CE2 PHE A 70 43 .929 8. 440 -27. 412

497 CD2 PHE A 70 43 .786 7. 578 -28. 495

498 C PHE A 70 43 .700 4. 274 -28. 989

499 0 PHE A 70 43 .737 4. 063 -27. 776

500 N CYS A 71 44 .769 4. 173 -29. 778

501 CA CYS A 71 46 .083 3. 775 -29. 264

502 CB CYS A 71 46 .940 3. 167 -30. 377

503 SG CYS A 71 46 .267 1. 648 -31. 065

504 C CYS A 71 46 .823 4. 938 -28. 614

505 0 CYS A 71 47 .006 5. 990 -29. 232

506 N ALA A 72 47 .242 4. 735 -27. 365

507 CA ALA A 72 47 .996 5. 740 -26. 613

508 CB ALA A 72 47 .075 6. 882 -26. 173

509 C ALA A 72 48 .702 5. 141 -25. 400

510 0 ALA A 72 48 .383 4. 033 -24. 964

511 N ALA A 73 49 .661 5. 888 -24. 861

512 CA ALA A 73 50 .286 5. 555 -23. 586

513 CB ALA A 73 51 .384 6. 559 -23. 264

514 C ALA A 73 49 .237 5. 533 -22. 469

515 0 ALA A 73 48 .446 6. 472 -22. 335

516 N CYS A 74 49 .241 4. 463 -21. 676

517 CA CYS A 74 48 .285 4. 284 -20. 579

518 CB CYS A 74 48 .699 3. 111 -19. 686

519 SG CYS A 74 47 .387 2. 526 -18. 567

520 C CYS A 74 48 .107 5. 542 -19. 724

521 0 CYS A 74 46 .979 5. 911 -19. 385

522 N HIS A 75 49 .217 6. 196 -19. 386

523 CA HIS A 75 49 .194 7. 351 -18. 489

524 CB HIS A 75 50 .564 7. 569 -17. 849

525 CG HIS A 75 50 .891 6. 558 -16. 797

526 ND1 HIS A 75 51 .503 5. 357 -17. 085

527 CE1 HIS A 75 51 .652 4. 664 -15. 969

528 NE2 HIS A 75 51 .153 5. 370 -14. 970

529 CD2 HIS A 75 50 .664 6. 556 -15. 462

530 C HIS A 75 48 .680 8. 622 -19. 151 531 0 HIS A 75 48..224 9.540 -18.464

532 N GLY A 76 48. .757 8 .668 -20 .480

533 CA GLY A 76 48. .130 9 .733 -21 .251

534 C GLY A 76 46, .627 9 .516 -21 .285

535 0 GLY A 76 45. .844 10 .467 -21 .206

536 N CYS A 77 46. .234 8 .251 -21 .390

537 CA CYS A 77 44. .832 7 .870 -21 .394

538 CB CYS A 77 44. .681 6 .399 -21 .765

539 SG CYS A 77 42. .963 5 .887 -21 .900

540 C CYS A 77 44. .186 8 .138 -20 .039

541 0 CYS A 77 43. .038 8 .586 -19 .964

542 N LEU A 78 44. .932 7 .874 -18 .972

543 CA LEU A 78 44. .433 8 .085 -17 .618

544 CB LEU A 78 45. .379 7 .455 -16 .590

545 CG LEU A 78 44. .978 7 .516 -15 . Ill

546 GDI LEU A 78 43. .766 6 .653 -14 .798

547 CD2 LEU A 78 46. .156 7 .149 -14 .227

548 C LEU A 78 44. .217 9 .571 -17 .324

549 0 LEU A 78 43. .266 9 .937 -16 .626

550 N PHE A 79 45. .096 10 .412 -17 .867

551 CA PHE A 79 45. .042 11 .860 -17 .649

552 CB PHE A 79 46. .326 12 .530 -18 .158

553 CG PHE A 79 46. .481 13 .961 -17 .721

554 CD1 PHE A 79 47. .038 14 .268 -16 .482

555 CE1 PHE A 79 47. .183 15 .595 -16 .068

556 CZ PHE A 79 46. .771 16 .630 -16 .903

557 CE2 PHE A 79 46. .214 16 .337 -18 .148

558 CD2 PHE A 79 46. .074 15 .005 -18 .550

559 C PHE A 79 43. .805 12 .502 -18 .289

560 0 PHE A 79 43. .222 13 .434 -17 .733

561 N ILE A 80 43. .415 11 .994 -19 .454

562 CA ILE A 80 42. .216 12 .461 -20 .141

563 CB ILE A 80 42. .180 11 .958 -21 .608

564 CGI ILE A 80 43. .388 12 .492 -22 .384

565 CD1 ILE A 80 43. .777 11 .643 -23 .589

566 CG2 ILE A 80 40. .889 12 .377 -22 .301

567 C ILE A 80 40. .949 12 .039 -19 .378

568 0 ILE A 80 40. .022 12 .837 -19 .213

569 N ALA A 81 40. .919 10 .792 -18 .912

570 CA ALA A 81 39. .802 10 .290 -18 .116

571 CB ALA A 81 39. .961 8 .804 -17 .853

572 C ALA A 81 39. .689 11 .066 -16 .804

573 0 ALA A 81 38. .588 11 .365 -16 .336

574 N CYS A 82 40. .839 11 .413 -16 .233

575 CA CYS A 82 40. .884 12 .197 -15 .007

576 CB CYS A 82 42. .300 12 .222 -14 .435

577 SG CYS A 82 42. .672 10 .768 -13 .436

578 C CYS A 82 40. .384 13 .608 -15 .222

579 0 CYS A 82 39. ,796 14 .208 -14 .319

580 N PHE A 83 40. .621 14 .136 -16 .419

581 CA PHE A 83 40. .204 15 .489 -16 .743

582 CB PHE A 83 40. .996 16 .044 -17 .928

583 CG PHE A 83 41. .359 17 .497 -17 .779

584 CD1 PHE A 83 42. .135 17 .927 -16 .698

585 CE1 PHE A 83 42. .470 19 .274 -16 .545

586 CZ PHE A 83 42. .032 20 .208 -17 .487

587 CE2 PHE A 83 41. .260 19 .789 -18 .573

588 CD2 PHE A 83 40. , 926 18 .438 -18 .710

589 C PHE A 83 38. .695 15 .559 -16 .973

590 0 PHE A 83 38. .066 16 .568 -16 .664 591 N VAL A 84 38..114 14.478 -17.488

592 CA VAL A 84 36. .658 14. 365 -17. 583

593 CB VAL A 84 36. .216 13. 094 -18. 366

594 CGI VAL A 84 34. .709 12. 926 -18. 325

595 CG2 VAL A 84 36. .673 13. 165 -19. 814

596 C VAL A 84 36. .033 14. 385 -16. 179

597 0 VAL A 84 34. .999 15. 025 -15. 962

598 N LEU A 85 36. .684 13. 701 -15. 237

599 CA LEU A 85 36. .205 13. 582 -13. 864

600 CB LEU A 85 36. .986 12. 495 -13. 114

601 CG LEU A 85 36. .693 11. 035 -13. 498

602 CD1 LEU A 85 37. .834 10. 107 -13. 086

603 CD2 LEU A 85 35. .348 10. 541 -12. 935

604 C LEU A 85 36. .245 14. 906 -13. 101

605 0 LEU A 85 35. .264 15. 274 -12. 451

606 N VAL A 86 37. .370 15. 618 -13. 192

607 CA VAL A 86 37. .535 16. 928 -12. 542

608 CB VAL A 86 38. .962 17. 516 -12. 778

609 CGI VAL A 86 39. .061 18. 974 -12. 295

610 CG2 VAL A 86 40. .018 16. 662 -12. 095

611 C VAL A 86 36. .486 17. 953 -12. 997

612 0 VAL A 86 35. .883 18. 639 -12. 166

613 N LEU A 87 36. .283 18. 050 -14. 311

614 CA LEU A 87 35. .411 19. 074 -14. 897

615 CB LEU A 87 35. .656 19. 218 -16. 407

616 CG LEU A 87 37. .024 19. 718 -16. 897

617 CD1 LEU A 87 37. .084 19. 660 -18. 416

618 CD2 LEU A 87 37. .355 21. 126 -16. 395

619 C LEU A 87 33. , 931 18. 819 -14. 616

620 0 LEU A 87 33. .176 19. 758 -14. 320

621 N THR A 88 33. .519 17. 556 -14. 702

622 CA THR A 88 32. .162 17. 190 -14. 326

623 CB THR A 88 31. .786 15. 760 -14. 780

624 0G1 THR A 88 32. .632 14. 796 -14. 138

625 CG2 THR A 88 31. .916 15. 629 -16. 297

626 C THR A 88 31. .935 17. 366 -12. 819

627 0 THR A 88 30. .827 17. 701 -12. 390

628 N ALA A 89 32. .987 17. 162 -12. 029

629 CA ALA A 89 32. .910 17. 329 -10. 578

630 CB ALA A 89 34. .173 16. 793 -9. 894

631 C ALA A 89 32. .672 18. 785 -10. 200

632 0 ALA A 89 31. .819 19. 074 -9. 351

633 N SER A 90 33. .421 19. 689 -10. 839

634 CA SER A 90 33. .271 21. 134 -10. 630

635 CB SER A 90 34. .279 21. 917 -11. 479

636 OG SER A 90 34. .012 21. 763 -12. 868

637 C SER A 90 31. .847 21. 581 -10. 945

638 0 SER A 90 31. .262 22. 379 -10. 218

639 N SER A 91 31, .303 21. 039 -12. 025

640 CA SER A 91 29. .909 21. 247 -12. 409

641 CB SER A 91 29. .605 20. 443 -13. 683

642 OG SER A 91 28. .213 20. 288 -13. 884

643 C SER A 91 28. .938 20. 870 -11. 277

644 0 SER A 91 28. .027 21. 630 -10. 960

645 N ILE A 92 29. .149 19. 701 -10. 673

646 CA ILE A 92 28. .328 19. 218 -9. 560

647 CB ILE A 92 28. .668 17. 744 -9. 207

648 CGI ILE A 92 28. .226 16. 800 -10. 334

649 CD1 ILE A 92 28. .822 15. 402 -10. 238

650 CG2 ILE A 92 28. .025 17. 339 -7. 887 651 C ILE A 92 28..472 20..091 -8.313

652 0 ILE A 92 27. .480 20. .464 -7. 695

653 N PHE A 93 29. .708 20. .422 -7. 953

654 CA PHE A 93 29. .975 21. .166 -6. 721

655 CB PHE A 93 31, .457 21, .098 -6. 345

656 CG PHE A 93 31. .893 19. .754 -5. 842

657 CD1 PHE A 93 32. .982 19, .107 -6. 412

658 CE1 PHE A 93 33. .394 17. .862 -5. 953

659 CZ PHE A 93 32. .707 17. .250 -4. 916

660 CE2 PHE A 93 31. .615 17, .887 -4. 336

661 CD2 PHE A 93 31. .211 19, .131 -4. 803

662 C PHE A 93 29. .515 22, .608 -6. 784

663 0 PHE A 93 29. .012 23, .144 -5. 792

664 N SER A 94 29. .693 23, .243 -7. 940

665 CA SER A 94 29. .225 24 , .614 -8. 112

666 CB SER A 94 29. .862 25, .263 -9. 344

667 OG SER A 94 29. .462 24 , .592 -10. 520

668 C SER A 94 27. .687 24. .676 -8. 161

669 0 SER A 94 27. .092 25, .705 -7. 840

670 N LEU A 95 27. .055 23. .572 -8. 557

671 CA LEU A 95 25. .606 23. .449 -8. 483

672 CB LEU A 95 25. .109 22. .266 -9. 316

673 CG LEU A 95 23. .589 22. .076 -9. 384

674 CD1 LEU A 95 22. .908 23. .249 -10. 111

675 CD2 LEU A 95 23. .236 20. .742 -10. 038

676 C LEU A 95 25. .149 23. .318 -7. 030

677 0 LEU A 95 24. .136 23. .908 -6. 639

678 N LEU A 96 25. .904 22. .555 -6. 239

679 CA LEU A 96 25. .678 22. .458 -4. 799

680 CB LEU A 96 26. .680 21. .479 -4. 164

681 CG LEU A 96 26. .652 21. .240 -2. 647

682 CD1 LEU A 96 25. .337 20. .596 -2. 205

683 CD2 LEU A 96 27. .832 20. .387 -2. 197

684 C LEU A 96 25, .749 23. .826 -4. 104

685 0 LEU A 96 24 , .943 24. .120 -3. 210

686 N ALA A 97 26. .710 24. .656 -4. 513

687 CA ALA A 97 26. .887 25. .990 -3. 924

688 CB ALA A 97 28. .074 26. .712 -4. 551

689 C ALA A 97 25. .619 26. .838 -4. 037

690 0 ALA A 97 25. .278 27. .579 -3. 116

691 N ILE A 98 24. .921 26. .715 -5. 163

692 CA ILE A 98 23. .639 27. .392 -5. 353

693 CB ILE A 98 23. .087 27. .185 -6. 785

694 CGI ILE A 98 24. .047 27. .814 -7. 803

695 CD1 ILE A 98 23. .867 27. .327 -9. 230

696 CG2 ILE A 98 21. .685 27. .790 -6. 926

697 C ILE A 98 22, .633 26. .957 -4. 280

698 0 ILE A 98 21. .986 27. .799 -3. 658

699 N ALA A 99 22. .543 25, .650 -4. 043

700 CA ALA A 99 21, .637 25. .091 -3. 037

701 CB ALA A 99 21. .606 23. .577 -3. 133

702 C ALA A 99 21. .945 25. .532 -1. 601

703 0 ALA A 99 21. .025 25. .840 -0. 843

704 N ILE A 100 23. .225 25. , 558 -1. 230

705 CA ILE A 100 23. ,640 26, .006 0. 108

706 CB ILE A 100 25. .138 25. ,710 0. 372

707 CGI ILE A 100 25. .371 24. ,198 0. 488

708 CD1 ILE A 100 26. .844 23. ,780 0. 466

709 CG2 ILE A 100 25. .619 26. ,408 1. 644

710 C ILE A 100 23. ,335 27. ,496 0. 294 711 0 ILE A 100 22,.914 27.932 1.375

712 N ASP A 101 23. .539 28. 262 -0. 777

713 CA ASP A 101 23. .214 29. 684 -0. 813

714 CB ASP A 101 23. .621 30. 278 -2. 161

715 CG ASP A 101 23. .258 31. 740 -2. 285

716 OD1 ASP A 101 23. , 831 32. 559 -1. 537

717 OD2 ASP A 101 22. .400 32. 069 -3. 133

718 C ASP A 101 21. , 725 29. 934 -0. 546

719 0 ASP A 101 21. .373 30. 736 0. 321

720 N ARG A 102 20. .866 29. 238 -1. 292

721 CA ARG A 102 19, .413 29. 332 -1. 121

722 CB ARG A 102 18. .672 28. 554 -2. 217

723 CG ARG A 102 18. .902 29. 054 -3. 648

724 CD ARG A 102 18. .302 30. 429 -3. 898

725 NE ARG A 102 19. .146 31. 507 -3. 388

726 CZ ARG A 102 18. .687 32. 652 -2. 890

727 NHl ARG A 102 17. .382 32. 882 -2. 818

728 NH2 ARG A 102 19. .538 33. 567 -2. 452

729 C ARG A 102 18. .964 28. 846 0. 254

730 0 ARG A 102 17. .973 29. 331 0. 789

731 N TYR A 103 19. .699 27. 888 0. 813

732 CA TYR A 103 19. .444 27. 396 2. 157

733 CB TYR A 103 20. .366 26. 218 2. 472

734 CG TYR A 103 20. .161 25. 611 3. 838

735 CD1 TYR A 103 21. .146 25. 708 4. 820

736 CE1 TYR A 103 20. .962 25. 144 6. 082

737 CZ TYR A 103 19. .778 24. 481 6. 367

738 OH TYR A 103 19. .579 23. 921 7. 607

739 CE2 TYR A 103 18. .788 24. 374 5. 409

740 CD2 TYR A 103 18. .983 24. 937 4. 152

741 C TYR A 103 19. .616 28. 518 3. 183

742 0 TYR A 103 18. .702 28. 798 3. 958

743 N ILE A 104 20. .780 29. 166 3. 162

744 CA ILE A 104 21. .079 30. 288 4. 055

745 CB ILE A 104 22. .536 30. 795 3. 866

746 CGI ILE A 104 23. .534 29. 653 4. 078

747 CD1 ILE A 104 24. .853 29. 840 3. 357

748 CG2 ILE A 104 22. .848 31. 953 4. 822

749 C ILE A 104 20. ,084 31. 433 3. 848

750 0 ILE A 104 19. ,586 32. Oil 4. 816

751 N ALA A 105 19. .786 31. 735 2. 586

752 CA ALA A 105 18. ,882 32. 831 2. 230

753 CB ALA A 105 18. ,822 33. 003 0. 723

754 C ALA A 105 17. ,474 32. 671 2. 801

755 0 ALA A 105 16. ,860 33. 653 3. 224

756 N ILE A 106 16. , 969 31. 439 2. 807

757 CA ILE A 106 15. , 612 31. 160 3. 276

758 CB ILE A 106 14. , 949 29. 981 2. 485

759 CGI ILE A 106 14. ,198 30. 504 1. 252

760 CD1 ILE A 106 15. ,064 30. 772 0. 027

761 CG2 ILE A 106 13. .974 29. 183 3. 355

762 C ILE A 106 15. ,562 30. 946 4. 790

763 0 ILE A 106 14. .643 31. 428 5. 454

764 N ARG A 107 16. .557 30. 250 5. 334

765 CA ARG A 107 16. .568 29. 940 6. 763

766 CB ARG A 107 17. ,368 28. 663 7. 051

767 CG ARG A 107 16. ,821 27. 397 6. 369

768 CD ARG A 107 15. ,389 27. 042 6. 799

769 NE ARG A 107 15. ,326 26. 491 8. 153

770 CZ ARG A 107 14. ,288 25. 826 8. 656 771 NH1 ARG A 107 13.202 25.610 7.923

772 NH2 ARG A 107 14 .339 25. 369 9. 900

773 C ARG A 107 17 .042 31. 096 7. 644

774 0 ARG A 107 16 .446 31. 361 8. 689

775 N ILE A 108 18 .102 31. 786 7. 220

776 CA ILE A 108 18 .621 32. 955 7. 947

111 CB ILE A 108 20 .060 32. 711 8. 490

778 CGI ILE A 108 20 .207 31. 288 9. 047

779 CD1 ILE A 108 21 .616 30. 719 8. 947

780 CG2 ILE A 108 20 .424 33. 756 9. 550

781 C ILE A 108 18 .603 34. 193 7. 035

782 0 ILE A 108 19 .657 34. 639 6. 569

783 N PRO A 109 17 .403 34. 756 6. 778

784 CA PRO A 109 17 .279 35. 819 5. 777

785 CB PRO A 109 15 .761 35. 920 5. 547

786 CG PRO A 109 15 .140 34. 801 6. 347

787 CD PRO A 109 16 .111 34. 488 7. 430

788 C PRO A 109 17 .830 37. 160 6. 250

789 0 PRO A 109 18 .153 38. 018 5. 424

790 N LEU A 110 17 .939 37. 328 7. 566

791 CA LEU A 110 18 .435 38. 565 8. 165

792 CB LEU A 110 18 .134 38. 593 9. 672

793 CG LEU A 110 16 .675 38. 698 10. 140

794 CD1 LEU A 110 16 .543 38. 289 11. 602

795 CD2 LEU A 110 16 .108 40. 101 9. 920

796 C LEU A 110 19 .930 38. 777 7. 908

797 0 LEU A 110 20 .317 39. 741 7. 244

798 N ARG A 111 20 .760 37. 868 8. 420

799 CA ARG A 111 22 .218 37. 995 8. 304

800 CB ARG A 111 22 .930 37. 246 9. 438

801 CG ARG A 111 23 .364 38. 152 10. 583

802 CD ARG A 111 24 .423 37. 493 11. 459

803 NE ARG A 111 23 .846 36. 535 12. 402

804 CZ ARG A 111 23 .431 36. 840 13. 629

805 NH1 ARG A 111 23 .519 38. 085 14. 082

806 NH2 ARG A 111 22 .923 35. 894 14. 407

807 C ARG A 111 22 .800 37. 588 6. 948

808 0 ARG A 111 23 .981 37. 826 6. 688

809 N TYR A 112 21 .971 36. 993 6. 090

810 CA TYR A 112 22 .402 36. 545 4. 763

811 CB TYR A 112 21 .197 36. 166 3. 891

812 CG TYR A 112 21 .565 35. 845 2. 455

813 CD1 TYR A 112 22 .017 34. 573 2. 100

814 CE1 TYR A 112 22 .359 34. 271 0. 786

815 CZ TYR A 112 22 .261 35. 249 -0. 187

816 OH TYR A 112 22 .602 34. 949 -1. 483

817 CE2 TYR A 112 21 .821 36. 524 0. 138

818 CD2 TYR A 112 21 .475 36. 815 1. 455

819 C TYR A 112 23 .275 37. 557 4. 018

820 0 TYR A 112 24 .299 37. 189 3. 441

821 N ASN A 113 22 .861 38. 821 4. 027

822 CA ASN A 113 23 .551 39. 870 3. 275

823 CB ASN A 113 22 .633 41. 079 3. 079

824 CG ASN A 113 21 .398 40. 740 2. 257

825 ODl ASN A 113 20 .309 40. 554 2. 800

826 ND2 ASN A 113 21 .570 40. 634 0. 944

827 C ASN A 113 24 .889 40. 276 3. 890

828 0 ASN A 113 25 .771 40. 783 3. 193

829 N GLY A 114 25 .031 40. 041 5. 193

830 CA GLY A 114 26 .290 40. 270 5. 899 831 C GLY A 114 27.197 39.050 5.893

832 0 GLY A 114 28 .420 39. 181 5. 956

833 N LEU A 115 26 .594 37. 863 5. 815

834 CA LEU A 115 27 .330 36. 598 5. 806

835 CB LEU A 115 26 .470 35. 472 6. 393

836 CG LEU A 115 27 .210 34. 245 6. 933

837 CD1 LEU A 115 27 .766 34. 512 8. 331

838 CD2 LEU A 115 26 .288 33. 043 6. 951

839 C LEU A 115 27 .828 36. 208 4. 408

840 0 LEU A 115 29 .014 35. 934 4. 223

841 N VAL A 116 26 .923 36. 182 3. 434

842 CA VAL A 116 27 .272 35. 789 2. 069

843 CB VAL A 116 26 .122 35. 001 1. 378

844 CGI VAL A 116 26 .545 34. 515 -0. 009

845 CG2 VAL A 116 25 .693 33. 821 2. 238

846 C VAL A 116 27 .696 37. 005 1. 240

847 0 VAL A 116 26 .908 37. 556 0. 465

848 N THR A 117 28 .952 37. 414 1. 419

849 CA THR A 117 29 .524 38. 554 0. 696

850 CB THR A 117 30 .562 39. 306 1. 552

851 0G1 THR A 117 31 .719 38. 482 1. 736

852 CG2 THR A 117 29 .982 39. 679 2. 912

853 C THR A 117 30 .190 38. 112 -0. 607

854 0 THR A 117 30 .387 36. 918 -0. 834

855 N GLY A 118 30 .543 39. 082 -1. 450

856 CA GLY A 118 31 .215 38. 817 -2. 723

857 C GLY A 118 32 .557 38. 121 -2. 579

858 0 GLY A 118 32 .933 37. 306 -3. 424

859 N THR A 119 33 .278 38. 445 -1. 508

860 CA THR A 119 34 .577 37. 836 -1. 215

861 CB THR A 119 35 .394 38. 711 -0. 223

862 OG1 THR A 119 36 .090 39. 723 -0. 958

863 CG2 THR A 119 36 .411 37. 891 0. 568

864 C THR A 119 34 .427 36. 393 -0. 718

865 0 THR A 119 35 .149 35. 499 -1. 165

866 N ARG A 120 33 .480 36. 175 0. 193

867 CA ARG A 120 33 .175 34. 835 0. 689

868 CB ARG A 120 32 .248 34. 901 1. 904

869 CG ARG A 120 33 .008 34. 821 3. 215

870 CD ARG A 120 32 .144 35. 059 4. 431

871 NE ARG A 120 32 .294 36. 421 4. 935

872 CZ ARG A 120 32 .073 36. 789 6. 195

873 NH1 ARG A 120 32 .242 38. 054 6. 551

874 NH2 ARG A 120 31 .693 35. 897 7. 104

875 C ARG A 120 32 .613 33. 912 -0. 392

876 0 ARG A 120 32 .849 32. 708 -0. 357

877 N ALA A 121 31 .891 34. 484 -1. 355

878 CA ALA A 121 31 .373 33. 728 -2. 500

879 CB ALA A 121 30 .424 34. 585 -3. 331

880 C ALA A 121 32 .500 33. 179 -3. 374

881 0 ALA A 121 32 .385 32. 087 -3. 926

882 N LYS A 122 33 .584 33. 941 -3. 494

883 CA LYS A 122 34 .759 33. 505 -4. 259

884 CB LYS A 122 35 .651 34. 694 -4. 609

885 CG LYS A 122 35 .059 35. 634 -5. 643

886 CD LYS A 122 35 .875 36. 901 -5. 736

887 CE LYS A 122 35 .215 37. 911 -6. 642

888 NZ LYS A 122 35 .617 39. 289 -6. 258

889 C LYS A 122 35 .552 32. 447 -3. 490

890 0 LYS A 122 36 .160 31. 560 -4. 090 891 N GLY A 123 35.534 32.553 -2.162

892 CA GLY A 123 36 .048 31. 506 -1. 291

893 C GLY A 123 35 .387 30. 176 -1. 615

894 0 GLY A 123 36 .074 29. 195 -1. 916

895 N ILE A 124 34 .052 30. 156 -1. 578

896 CA ILE A 124 33 .271 28. 955 -1. 882

897 CB ILE A 124 31 .743 29. 169 -1. 648

898 CGI ILE A 124 31 .385 28. 947 -0. 179

899 CD1 ILE A 124 31 .432 30. 186 0. 679

900 CG2 ILE A 124 30 .919 28. 187 -2. 454

901 C ILE A 124 33 .563 28. 422 -3. 289

902 0 ILE A 124 33 .756 27. 216 -3. 466

903 N ILE A 125 33 .611 29. 319 -4. 274

904 CA ILE A 125 33 .943 28. 941 -5. 653

905 CB ILE A 125 33 .970 30. 170 -6. 593

906 CGI ILE A 125 32 .575 30. 796 -6. 696

907 CD1 ILE A 125 32 .565 32. 227 -7. 246

908 CG2 ILE A 125 34 .521 29. 796 -7. 989

909 C ILE A 125 35 .286 28. 202 -5. 732

910 0 ILE A 125 35 .377 27. 144 -6. 363

911 N ALA A 126 36 .314 28. 769 -5. 092

912 CA ALA A 126 37 .654 28. 176 -5. 055

913 CB ALA A 126 38 .629 29. 098 -4. 330

914 C ALA A 126 37 .646 26. 782 -4. 415

915 0 ALA A 126 38 .230 25. 842 -4. 962

916 N ILE A 127 36 .970 26. 662 -3. 272

917 CA ILE A 127 36 .742 25. 379 -2. 615

918 CB ILE A 127 35 .886 25. 544 -1. 324

919 CGI ILE A 127 36 .701 26. 265 -0. 244

920 CD1 ILE A 127 35 .883 26. 920 0. 875

921 CG2 ILE A 127 35 .399 24. 196 -0. 809

922 C ILE A 127 36 .128 24. 342 -3. 575

923 0 ILE A 127 36 .560 23. 187 -3. 592

924 N CYS A 128 35 .152 24. 756 -4. 383

925 CA CYS A 128 34 .540 23. 858 -5. 374

926 CB CYS A 128 33 .446 24. 572 -6. 179

927 SG CYS A 128 31 .900 24. 794 -5. 284

928 C CYS A 128 35 .569 23. 271 -6. 329

929 0 CYS A 128 35 .496 22. 091 -6. 690

930 N TRP A 129 36 .526 24. 104 -6. 725

931 CA TRP A 129 37 .570 23. 693 -7. 651

932 CB TRP A 129 38 .096 24. 893 -8. 424

933 CG TRP A 129 37 .213 25. 237 -9. 575

934 CD1 TRP A 129 36 .235 26. 191 -9. 607

935 NE1 TRP A 129 35 .632 26. 209 -10. 842

936 CE2 TRP A 129 36 .210 25. 252 -11. 632

937 CD2 TRP A 129 37 .210 24. 614 -10. 862

938 CE3 TRP A 129 37 .961 23. 581 -11. 445

939 CZ3 TRP A 129 37 .691 23. 223 -12. 763

940 CH2 TRP A 129 36 .690 23. 884 -13. 503

941 CZ2 TRP A 129 35 .943 24. 895 -12. 954

942 C TRP A 129 38 .712 22. 919 -6. 999

943 0 TRP A 129 39 .315 22. 064 -7. 641

944 N VAL A 130 39 .005 23. 211 -5. 736

945 CA VAL A 130 40 .012 22. 445 -5. 001

946 CB VAL A 130 40 .347 23. 076 -3. 624

947 CGI VAL A 130 41 .123 22. 093 -2. 734

948 CG2 VAL A 130 41 .129 24. 373 -3. 807

949 C VAL A 130 39 .533 20. 999 -4. 851

950 0 VAL A 130 40 .268 20. 065 -5. 185 951 N LEU A 131 38.296 20.831 -4.381

952 CA LEU A 131 37 .684 19 .513 -4. 237

953 CB LEU A 131 36 .295 19 .623 -3. 597

954 CG LEU A 131 36 .110 19 .589 -2. 066

955 CD1 LEU A 131 36 .551 18 .250 -1. 476

956 CD2 LEU A 131 36 .810 20 .733 -1. 336

957 C LEU A 131 37 .596 18 .746 -5. 562

958 0 LEU A 131 37 .730 17 .527 -5. 582

959 N SER A 132 37 .380 19 .460 -6. 664

960 CA SER A 132 37 .288 18 .832 -7. 981

961 CB SER A 132 36 .775 19 .826 -9. 024

962 OG SER A 132 35 .500 20 .313 -8. 649

963 C SER A 132 38 .615 18 .230 -8. 429

964 0 SER A 132 38 .644 17 .104 -8. 921

965 N PHE A 133 39 .703 18 .984 -8. 251

966 CA PHE A 133 41 .051 18 .529 -8. 605

967 CB PHE A 133 42 .062 19 .687 -8. 554

968 CG PHE A 133 42 .077 20 .547 -9. 798

969 CD1 PHE A 133 41 .554 21 .837 -9. 777

970 CE1 PHE A 133 41 .563 22 .641 -10. 928

971 CZ PHE A 133 42 .099 22 .149 -12. 112

972 CE2 PHE A 133 42 .625 20 .860 -12. 147

973 CD2 PHE A 133 42 .614 20 .066 -10. 990

974 C PHE A 133 41 .506 17 .386 -7. 706

975 0 PHE A 133 41 .934 16 .343 -8. 191

976 N ALA A 134 41 .391 17 .581 -6. 394

977 CA ALA A 134 41 .778 16 .566 -5. 421

978 CB ALA A 134 41 .441 17 .027 -4. 023

979 C ALA A 134 41 .131 15 .206 -5. 707

980 0 ALA A 134 41 .775 14 .166 -5. 553

981 N ILE A 135 39 .868 15 .220 -6. 132

982 CA ILE A 135 39 .102 13 .990 -6. 316

983 CB ILE A 135 37 .592 14 .212 -6. 014

984 CGI ILE A 135 37 .398 14 .471 -4. 514

985 CD1 ILE A 135 36 .092 15 .155 -4. 142

986 CG2 ILE A 135 36 .747 13 .004 -6. 466

987 C ILE A 135 39 .312 13 .380 -7. 704

988 0 ILE A 135 39 .559 12 .181 -7. 833

989 N GLY A 136 39 .217 14 .213 -8. 734

990 CA GLY A 136 39 .348 13 .754 -10. 102

991 C GLY A 136 40 .764 13 .399 -10. 491

992 0 GLY A 136 40 .961 12 .625 -11. 426

993 N LEU A 137 41 .746 13 .965 -9. 790

994 CA LEU A 137 43 .157 13 .650 -10. 037

995 CB LEU A 137 44 .053 14 .893 -9. 902

996 CG LEU A 137 43 .971 16 .038 -10. 918

997 CD1 LEU A 137 45 .025 17 .089 -10. 576

998 CD2 LEU A 137 44 .128 15 .561 -12. 366

999 C LEU A 137 43 .704 12 .540 -9. 133

1000 0 LEU A 137 44 .849 12 .111 -9. 314

1001 N THR A 138 42 .902 12 .092 -8. 163

1002 CA THR A 138 43 .310 11 .021 -7. 244

1003 CB THR A 138 42 .164 10 .618 -6. 258

1004 OG1 THR A 138 42 .084 11 .584 -5. 207

1.005 CG2 THR A 138 42 .404 9 .251 -5. 624

1006 C THR A 138 43 .908 9 .799 -7. 970

1007 0 THR A 138 44 .934 9 .271 -7. 525

1008 N PRO A 139 43 .293 9 .363 -9. 098

1009 CA PRO A 139 43 .887 8 .254 -9. 853

1010 CB PRO A 139 42 .911 8 .069 -11. 019 1011 CG PRO A 139 41.612 8.531 -10.472

1012 CD PRO A 139 41. 971 9. 726 -9. 648

1013 C PRO A 139 45. 309 8. 501 -10. 372

1014 0 PRO A 139 46. 045 7. 544 -10. 601

1015 N MET A 140 45. 690 9. 764 -10. 549

1016 CA MET A 140 47. 047 10. 120 -10. 980

1017 CB MET A 140 47. 162 11. 627 -11. 204

1018 CG MET A 140 46. 335 12. 177 -12. 351

1019 SD MET A 140 46. 998 11. 709 -13. 955

1020 CE MET A 140 45. 882 10. 408 -14. 385

1021 C MET A 140 48. 115 9. 693 -9. 977

1022 0 MET A 140 49. 229 9. 350 -10. 361

1023 N LEU A 141 47. 758 9. 716 -8. 696

1024 CA LEU A 141 48. 683 9. 416 -7. 603

1025 CB LEU A 141 48. 100 9. 885 -6. 261

1026 CG LEU A 141 47. 638 11. 336 -6. 108

1027 CD1 LEU A 141 46. 965 11. 524 -4. 753

1028 CD2 LEU A 141 48. 789 12. 323 -6. 292

1029 C LEU A 141 49. 059 7. 939 -7. 496

1030 0 LEU A 141 49. 931 7. 580 -6. 704

1031 N GLY A 142 48. 400 7. 084 -8. 270

1032 CA GLY A 142 48. 711 5. 660 -8. 245

1033 C GLY A 142 47. 504 4. 748 -8. 202

1034 0 GLY A 142 47. 613 3. 564 -8. 529

1035 N TRP A 143 46. 356 5. 288 -7. 793

1036 CA TRP A 143 45. 111 4. 522 -7. 774

1037 CB TRP A 143 44. 090 5. 172 -6. 834

1038 CG TRP A 143 42. 991 4. 242 -6. 390

1039 CD1 TRP A 143 42. 792 2. 954 -6. 796

1040 NE1 TRP A 143 41. 684 2. 427 -6. 183

1041 CE2 TRP A 143 41. 130 3. 379 -5. 370

1042 CD2 TRP A 143 41. 926 4. 542 -5. 477

1043 CE3 TRP A 143 41. 571 5. 675 -4. 731

1044 CZ3 TRP A 143 40. 445 5. 614 -3. 910

1045 CH2 TRP A 143 39. 672 4. 444 -3. 825

1046 CZ2 TRP A 143 39. 998 3. 316 -4. 543

1047 C TRP A 143 44. 545 4. 374 -9. 201

1048 0 TRP A 143 43. 541 5. 000 -9. 564

1049 N ASN A 144 45. 207 3. 546 -10. 008

1050 CA ASN A 144 44. 820 3. 346 -11. 406

1051 CB ASN A 144 45. 463 4. 405 -12. 300

1052 CG ASN A 144 46. 973 4. 409 -12. 203

1053 0D1 ASN A 144 47. 638 3. 542 -12. 758

1054 ND2 ASN A 144 47. 522 5. 395 -11. 503

1055 C ASN A 144 45. 144 1. 941 -11. 904

1056 0 ASN A 144 45. 786 1. 164 -11. 196

1057 N ASN A 145 44. 691 1. 621 -13. 117

1058 CA ASN A 145 44. 847 0. 278 -13. 682

1059 CB ASN A 145 43. 577 -0. 149 -14. 415

1060 CG ASN A 145 42. 454 -0. 533 -13. 480

1061 OD1 ASN A 145 42. 658 -0. 743 -12. 281

1062 ND2 ASN A 145 41. 250 -0. 635 -14. 028

1063 C ASN A 145 46. 034 0. 152 -14. 624

1064 0 ASN A 145 46. 202 -0. 878 -15. 279

1065 N CYS A 146 46. 860 1. 194 -14. 678

1066 CA CYS A 146 47. 971 1. 258 -15. 632

1067 CB CYS A 146 48. 501 2. 686 -15. 750

1068 SG CYS A 146 47. 534 3. 691 -16. 881

1069 C CYS A 146 49. 114 0. 280 -15. 367

1070 0 CYS A 146 49. 921 0. 006 -16. 260 1071 N GLY A 147 49.176 -0.245 -14.145

1072 CA GLY A 147 50. 141 -1 .280 -13. 796

1073 C GLY A 147 49. 829 -2 .596 -14. 483

1074 0 GLY A 147 50. 716 -3 .429 -14. 666

1075 N GLN A 148 48. 566 -2 .775 -14. 870

1076 CA GLN A 148 48. 090 -4 .027 -15. 461

1077 CB GLN A 148 47. 084 -4 .715 -14. 521

1078 CG GLN A 148 47. 653 -5 .152 -13. 165

1079 CD GLN A 148 47. 831 -3 .996 -12. 183

1080 OE1 GLN A 148 46. 902 -3 .220 -11. 937

1081 NE2 GLN A 148 49. 029 -3 .881 -11. 616

1082 C GLN A 148 47. 465 -3 .824 -16. 850

1083 0 GLN A 148 46. 261 -4 .034 -17. 021

1084 N PRO A 149 48. 286 -3 .437 -17. 853

1085 CA PRO A 149 47. 784 -3 .171 -19. 208

1086 CB PRO A 149 49. 028 -2 .683 -19. 967

1087 CG PRO A 149 50. 082 -2 .441 -18. 927

1088 CD PRO A 149 49. 757 -3 .363 -17. 804

1089 C PRO A 149 47. 249 -4 .431 -19. 874

1090 0 PRO A 149 47. 619 -5 .540 -19. 488

1091 N LYS A 150 46. 386 -4 .259 -20. 868

1092 CA LYS A 150 45. 796 -5 .393 -21. 573

1093 CB LYS A 150 44. 272 -5 .265 -21. 609

1094 CG LYS A 150 43. 665 -5 .312 -20. 221

1095 CD LYS A 150 42. 179 -5 .068 -20. 222

1096 CE LYS A 150 41. 657 -5 .094 -18. 799

1097 NZ LYS A 150 40. 188 -4 .955 -18. 761

1098 C LYS A 150 46. 393 -5 .542 -22. 968

1099 0 LYS A 150 46. 037 -4 .803 -23. 888

1100 N GLU A 151 47. 319 -6 .493 -23. 096

1101 CA GLU A 151 48. 058 -6 .735 -24. 337

1102 CB GLU A 151 49. 292 -7 .609 -24. 073

1103 CG GLU A 151 50. 594 -6 .833 -23. 869

1104 CD GLU A 151 50. 656 -6 .095 -22. 541

1105 OE1 GLU A 151 50. 183 -6 .646 -21. 523

1106 OE2 GLU A 151 51. 190 -4 .965 -22. 516

1107 C GLU A 151 47. 201 -7 .357 -25. 434

1108 0 GLU A 151 47. 329 -6 .991 -26. 603

1109 N GLY A 152 46. 337 -8 .295 -25. 050

1110 CA GLY A 152 45. 438 -8 .974 -25. 989

1111 C GLY A 152 44. 506 -8 .024 -26. 718

1112 0 GLY A 152 44. 276 -8 .168 -27. 920

1113 N LYS A 153 43. 977 -7 .050 -25. 981

1114 CA LYS A 153 43. 111 -6 .010 -26. 532

1115 CB LYS A 153 42. 276 -5 .372 -25. 412

1116 CG LYS A 153 41. 427 -4 .173 -25. 828

1117 CD LYS A 153 40. 078 -4 .141 -25. 119

1118 CE LYS A 153 40. 214 -4 .032 -23. 610

1119 NZ LYS A 153 38. 900 -3 .844 -22. 943

1120 C LYS A 153 43. 923 -4 .956 -27. 294

1121 0 LYS A 153 43. 467 -4 .432 -28. 316

1122 N ALA A 154 45. 121 -4 .655 -26. 792

1123 CA ALA A 154 46. 027 -3 .708 -27. 440

1124 CB ALA A 154 47. 223 -3 .415 -26. 548

1125 C ALA A 154 46. 489 -4 .227 -28. 800

1126 0 ALA A 154 46. 655 -3 .450 -29. 744

1127 N HIS A 155 46. 687 -5 .542 -28. 887

1128 CA HIS A 155 47. 082 -6 .195 -30. 130

1129 CB HIS A 155 47. 499 -7 .649 -29. 875

1130 CG HIS A 155 47. 978 -8 .366 -31. 100 1131 ND1 HIS A 155 49.116 -7.994 -31.785

1132 CE1 HIS A 155 49 .291 -8 .802 -32. 815

1133 NE2 HIS A 155 48 .309 -9 .686 -32. 822

1134 CD2 HIS A 155 47 .474 -9 .436 -31. 760

1135 C HIS A 155 45 .967 -6 .130 -31. 173

1136 0 HIS A 155 46 .213 -5 .760 -32. 323

1137 N SER A 156 44 .749 -6 .480 -30. 759

1138 CA SER A 156 43 .576 -6 .458 -31. 639

1139 CB SER A 156 42 .330 -6 .912 -30. 879

1140 OG SER A 156 42 .453 -8 .262 -30. 465

1141 C SER A 156 43 .327 -5 .096 -32. 297

1142 0 SER A 156 42 .834 -5 .031 -33. 423

1143 N GLN A 157 43 .677 -4 .019 -31. 595

1144 CA GLN A 157 43 .532 -2 .656 -32. 116

1145 CB GLN A 157 43 .269 -1 .662 -30. 974

1146 CG GLN A 157 41 .928 -1 .838 -30. 272

1147 CD GLN A 157 40 .745 -1 .622 -31. 193

1148 OE1 GLN A 157 40 .589 -0 .550 -31. 783

1149 NE2 GLN A 157 39 .895 -2 .638 -31. 312

1150 C GLN A 157 44 .749 -2 .207 -32. 930

1151 0 GLN A 157 44 .776 -1 .089 -33. 452

1152 N GLY A 158 45 .748 -3 .081 -33. 031

1153 CA GLY A 158 46 .975 -2 .789 -33. 767

1154 C GLY A 158 47 .799 -1 .667 -33. 165

1155 0 GLY A 158 48 .376 -0 .858 -33. 894

1156 N CYS A 159 47 .849 -1 .614 -31. 835

1157 CA CYS A 159 48 .652 -0 .616 -31. 131

1158 CB CYS A 159 48 .128 -0 .384 -29. 706

1159 SG CYS A 159 46 .435 0 .292 -29. 550

1160 C CYS A 159 50 .111 -1 .050 -31. 078

1161 0 CYS A 159 50 .409 -2 .233 -30. 897

1162 N GLY A 160 51 .014 -0 .086 -31. 235

1163 CA GLY A 160 52 .449 -0 .338 -31. 118

1164 C GLY A 160 52 .826 -0 .714 -29. 699

1165 0 GLY A 160 52 .035 -0 .528 -28. 772

1166 N GLU A 161 54 .034 -1 .246 -29. 527

1167 CA GLU A 161 54 .495 -1 .690 -28. 210

1168 CB GLU A 161 55 .837 -2 .430 -28. 304

1169 CG GLU A 161 56 .950 -1 .681 -29. 036

1170 CD GLU A 161 58 .132 -2 .571 -29. 400

1171 OE1 GLU A 161 58 .285 -3 .660 -28. 800

1172 OE2 GLU A 161 58 .912 -2 .176 -30. 292

1173 C GLU A 161 54 .561 -0 .536 -27. 212

1174 0 GLU A 161 55 .032 0 .556 -27. 541

1175 N GLY A 162 54 .067 -0 .784 -26. 002

1176 CA GLY A 162 53 .979 0 .249 -24. 972

1177 C GLY A 162 52 .750 1 .131 -25. 118

1178 0 GLY A 162 52 .714 2 .249 -24. 598

1179 N GLN A 163 51 .747 0 .630 -25. 836

1180 CA GLN A 163 50 .481 1 .338 -26. 011

1181 CB GLN A 163 50 .359 1 .912 -27. 423

1182 CG GLN A 163 51 .283 3 .083 -27. 714

1183 CD GLN A 163 50 .970 3 .753 -29. 040

1184 OE1 GLN A 163 50 .776 3 .087 -30. 060

1185 NE2 GLN A 163 50 .918 5 .081 -29. 031

1186 C GLN A 163 49 .293 0 .427 -25. 722

1187 O GLN A 163 49 .374 -0 .795 -25. 885

1188 N VAL A 164 48 .193 1 .037 -25. 289

1189 CA VAL A 164 46 .967 0 .311 -24. 966

1190 CB VAL A 164 46 .743 0 .190 -23. 428 1191 CGI VAL A 164 47.835 -0..656 -22.787

1192 CG2 VAL A 164 46 .656 1. .571 -22. 762

1193 C VAL A 164 45 .756 0. .985 -25. 600

1194 0 VAL A 164 45 .810 2. .170 -25. 947

1195 N ALA A 165 44 .674 0. ,222 -25. 750

1196 CA ALA A 165 43 .388 0. .767 -26. 173

1197 CB ALA A 165 42 .411 -0. .361 -26. 480

1198 C ALA A 165 42 .834 1. .686 -25. 081

1199 0 ALA A 165 42 .522 1. .243 -23. 973

1200 N CYS A 166 42 .730 2. .972 -25. 395

1201 CA CYS A 166 42 .307 3. .966 -24. 418

1202 CB CYS A 166 42 .703 5. .368 -24. 886

1203 SG CYS A 166 42 .385 6. .679 -23. 688

1204 C CYS A 166 40 .807 3, .892 -24. 105

1205 0 CYS A 166 39 .992 4 , .545 -24. 760

1206 N LEU A 167 40 .459 3. .078 -23. 108

1207 CA LEU A 167 39 .091 2. .990 -22. 596

1208 CB LEU A 167 38 .539 1. .567 -22. 722

1209 CG LEU A 167 38 .083 0. .930 -24. 046

1210 CD1 LEU A 167 37 .443 1. .929 -25. 014

1211 CD2 LEU A 167 39 .216 0. .166 -24. 712

1212 C LEU A 167 39 .027 3. .427 -21. 132

1213 0 LEU A 167 39 .982 3. .223 -20. 373

1214 N PHE A 168 37 .889 4 , .005 -20. 741

1215 CA PHE A 168 37 .687 4, .528 -19. 384

1216 CB PHE A 168 36 .312 5. .201 -19. 257

1217 CG PHE A 168 36 .092 5. .909 -17. 945

1218 CD1 PHE A 168 36 .452 7. .246 -17. 793

1219 CE1 PHE A 168 36 .252 7. .911 -16. 577

1220 CZ PHE A 168 35 .682 7. .231 -15. 497

1221 CE2 PHE A 168 35 .315 5. .894 -15. 639

1222 CD2 PHE A 168 35 .520 5. .240 -16. 860

1223 C PHE A 168 37 .847 3. .438 -18. 333

1224 0 PHE A 168 38 .642 3. .581 -17. 400

1225 N GLU A 169 37 .103 2. .349 -18. 506

1226 CA GLU A 169 37 .113 1. .223 -17. 566

1227 CB GLU A 169 35 .914 0. .298 -17. 814

1228 CG GLU A 169 34 .569 0. .931 -17. 494

1229 CD GLU A 169 33 .417 -0. .062 -17. 525

1230 0E1 GLU A 169 33 .671 -1. .286 -17. 518

1231 OE2 GLU A 169 32 .250 0. .385 -17. 550

1232 C GLU A 169 38 .410 0. .417 -17. 593

1233 0 GLU A 169 38 .552 -0. .552 -16. 852

1234 N ASP A 170 39 .351 0. .815 -18. 445

1235 CA ASP A 170 40 .652 0. .152 -18. 517

1236 CB ASP A 170 41 .041 -0. .118 -19. 974

1237 CG ASP A 170 40 .234 -1. .251 -20. 594

1238 OD1 ASP A 170 39 .450 -1. .908 -19. 870

1239 OD2 ASP A 170 40 .383 -1. .489 -21. 810

1240 C ASP A 170 41 .763 0. .903 -17. 780

1241 0 ASP A 170 42 .787 0. .318 -17. 444

1242 N VAL A 171 41 .559 2. .192 -17. 533

1243 CA VAL A 171 42 .534 2. .996 -16. 792

1244 CB VAL A 171 43 .042 4. .220 -17. 618

1245 CGI VAL A 171 43 .830 3. .752 -18. 838

1246 CG2 VAL A 171 41 .887 5. .133 -18. 042

1247 C VAL A 171 42 .007 3. ,440 -15. 415

1248 0 VAL A 171 42 .770 3. .503 -14. 452

1249 N VAL A 172 40 .711 3. ,735 -15. 326

1250 CA VAL A 172 40 .101 4. ,167 -14. 069 1251 CB VAL A 172 39.016 5.267 -14.286

1252 CGI VAL A 172 38. 366 5. 650 -12. 967

1253 CG2 VAL A 172 39. 623 6. 499 -14. 942

1254 C VAL A 172 39. 499 2. 960 -13. 341

1255 0 VAL A 172 38. 612 2. 296 -13. 877

1256 N PRO A 173 39. 976 2. 678 -12. 115

1257 CA PRO A 173 39. 502 1. 502 -11. 378

1258 CB PRO A 173 40. 441 1. 435 -10. 161

1259 CG PRO A 173 41. 614 2. 309 -10. 510

1260 CD PRO A 173 41. 038 3. 392 -11. 382

1261 C PRO A 173 38. 061 1. 641 -10. 917

1262 0 PRO A 173 37. 613 2. 740 -10. 596

1263 N MET A 174 37. 344 0. 524 -10. 889

1264 CA MET A 174 35. 960 0. 514 -10. 440

1265 CB MET A 174 35. 256 -0. 782 -10. 850

1266 CG MET A 174 34. 671 -0. 783 -12. 263

1267 SD MET A 174 33. 542 0. 578 -12. 681

1268 CE MET A 174 32. 624 0. 727 -11. 164

1269 C MET A 174 35. 779 0. 771 -8. 934

1270 0 MET A 174 34. 740 1. 289 -8. 519

1271 N ASN A 175 36. 765 0. 412 -8. 114

1272 CA ASN A 175 36. 636 0. 676 -6. 678

1273 CB ASN A 175 37. 569 -0. 193 -5. 817

1274 CG ASN A 175 39. 020 -0. 107 -6. 234

1275 0D1 ASN A 175 39. 375 0. 594 -7. 183

1276 ND2 ASN A 175 39. 874 -0. 830 -5. 522

1277 C ASN A 175 36. 756 2. 161 -6. 363

1278 0 ASN A 175 36. 009 2. 680 -5. 526

1279 N TYR A 176 37. 670 2. 838 -7. 063

1280 CA TYR A 176 37. 737 4. 301 -7. 049

1281 CB TYR A 176 38. 826 4. 832 -7. 996

1282 CG TYR A 176 38. 661 6. 313 -8. 253

1283 CD1 TYR A 176 38. 006 6. 768 -9. 396

1284 CE1 TYR A 176 37. 815 8. 123 -9. 624

1285 CZ TYR A 176 38. 272 9. 039 -8. 700

1286 OH TYR A 176 38. 077 10. 370 -8. 938

1287 CE2 TYR A 176 38. 925 8. 621 -7. 549

1288 CD2 TYR A 176 39. 108 7. 260 -7. 325

1289 C TYR A 176 36. 390 4. 947 -7. 408

1290 0 TYR A 176 35. 922 5. 855 -6. 716

1291 N MET A 177 35. 783 4. 473 -8. 493

1292 CA MET A 177 34. 496 4. 988 -8. 955

1293 CB MET A 177 34. 117 4. 376 -10. 315

1294 CG MET A 177 34. 864 4. 975 -11. 534

1295 SD MET A 177 34. 854 6. 786 -11. 607

1296 CE MET A 177 33. 093 7. 119 -11. 775

1297 C MET A 177 33. 376 4. 766 -7. 949

1298 0 MET A 177 32. 562 5. 658 -7. 712

1299 N VAL A 178 33. 354 3. 581 -7. 350

1300 CA VAL A 178 32. 275 3. 174 -6. 459

1301 CB VAL A 178 32. 106 1. 627 -6. 455

1302 CGI VAL A 178 31. 263 1. 148 -5. 270

1303 CG2 VAL A 178 31. 506 1. 158 -7. 772

1304 C VAL A 178 32. 455 3. 712 -5. 037

1305 0 VAL A 178 31. 512 4. 257 -4. 463

1306 N TYR A 179 33. 663 3. 577 -4. 490

1307 CA TYR A 179 33. 926 3. 911 -3. 082

1308 CB TYR A 179 35. 064 3. 055 -2. 512

1309 CG TYR A 179 34. 694 1. 610 -2. 279

1310 CD1 TYR A 179 35. 542 0. 581 -2. 682 1311 CE1 TYR A 179 35.,203 -0..753 -2..472

1312 cz TYR A 179 34. .001 -1. .067 -1. .857

1313 OH TYR A 179 33. .665 -2. .381 -1. .646

1314 CE2 TYR A 179 33. .139 -0. .066 -1. .448

1315 CD2 TYR A 179 33. .489 1. .268 -1. .660

1316 C TYR A 179 34. ,230 5. .376 -2. .825

1317 0 TYR A 179 33. .898 5. .904 -1. ,761

1318 N PHE A 180 34. .871 6. .028 -3. .788

1319 CA PHE A 180 35. .280 7. .412 -3. .612

1320 CB PHE A 180 36. .788 7 , .559 -3. .887

1321 CG PHE A 180 37. .370 8. .906 -3. .512

1322 CD1 PHE A 180 36. .764 9. .720 -2. .553

1323 CE1 PHE A 180 37. .316 10. .964 -2. .211

1324 CZ PHE A 180 38. .492 11. .387 -2. .815

1325 CE2 PHE A 180 39. .115 10. .572 -3. .765

1326 CD2 PHE A 180 38. .552 9. .342 -4. .104

1327 C PHE A 180 34 , .416 8. .360 -4. .456

1328 0 PHE A 180 33. .704 9. .199 -3. .900

1329 N ASN A 181 34 , .451 8. .199 -5. .779

1330 CA ASN A 181 33, .774 9. .119 -6. .704

1331 CB ASN A 181 34. .177 8. .817 -8. .150

1332 CG ASN A 181 33. .949 9. .992 -9. .075

1333 ODl ASN A 181 32. .826 10. .258 -9. .479

1334 ND2 ASN A 181 35. .022 10. .694 -9. .424

1335 C ASN A 181 32. .243 9. , 176 -6. .569

1336 0 ASN A 181 31. .664 10. .259 -6. .505

1337 N PHE A 182 31, .593 8. .018 -6. .521

1338 CA PHE A 182 30, .139 7. .963 -6. .333

1339 CB PHE A 182 29, .650 6. .508 -6. .443

1340 CG PHE A 182 28, .193 6. .309 -6. .117

1341 CD1 PHE A 182 27. .210 7. .120 -6. .677

1342 CE1 PHE A 182 25. .861 6. .921 -6. .380

1343 CZ PHE A 182 25. .486 5. .891 -5. ,521

1344 CE2 PHE A 182 26. .461 5. .066 -4. ,964

1345 CD2 PHE A 182 27. .802 5. .276 -5. ,268

1346 C PHE A 182 29, .675 8. .640 -5. .023

1347 0 PHE A 182 28, .709 9. .407 -5. .028

1348 N PHE A 183 30, .368 8. .372 -3. .917

1349 CA PHE A 183 29. .985 8. .937 -2. .619

1350 CB PHE A 183 30. .608 8. .139 -1. ,462

1351 CG PHE A 183 29. .961 6. .795 -1. .235

1352 CD1 PHE A 183 28. .774 6. .688 -0. ,508

1353 CE1 PHE A 183 28. .170 5. .443 -0. ,301

1354 CZ PHE A 183 28. .755 4. .286 -0. .828

1355 CE2 PHE A 183 29, .938 4. .382 -1. .550

1356 CD2 PHE A 183 30. .536 5. .634 -1. , 750

1357 C PHE A 183 30. .305 10. .429 -2. ,492

1358 0 PHE A 183 29. .458 11. .215 -2. , 060

1359 N ALA A 184 31. .517 10. .812 -2. ,882

1360 CA ALA A 184 31, .954 12. .202 -2. ,791

1361 CB ALA A 184 33. .465 12. .304 -2. ,988

1362 C ALA A 184 31. .233 13. , 119 -3. ,777

1363 0 ALA A 184 30. .854 14. ,239 -3. 423

1364 N CYS A 185 31. .047 12. .642 -5. ,006

1365 CA CYS A 185 30, .593 13. .498 -6. ,102

1366 CB CYS A 185 31. .368 13. ,194 -7. ,388

1367 SG CYS A 185 33. .120 13. , 605 -7. ,295

1368 C CYS A 185 29. .100 13. , 437 -6. ,382

1369 0 CYS A 185 28. .528 14. ,400 -6. 890

1370 N VAL A 186 28. , 471 12. ,311 -6. 071 1371 CA VAL A 186 27..076 12.129 -6.438

1372 CB VAL A 186 26. .889 10 .945 -7. 421

1373 CGI VAL A 186 25. , 401 10 .724 -7. 754

1374 CG2 VAL A 186 27. .712 11 .175 -8. 692

1375 C VAL A 186 26. .164 12 .001 -5. 218

1376 0 VAL A 186 25. .237 12 .798 -5. 053

1377 N LEU A 187 26. ,426 11 .012 -4. 369

1378 CA LEU A 187 25. .523 10 .736 -3. 253

1379 CB LEU A 187 25, .829 9 .389 -2. 607

1380 CG LEU A 187 24 , .628 8 .795 -1. 879

1381 CD1 LEU A 187 23. .666 8 .155 -2. 880

1382 CD2 LEU A 187 25. .079 7 .787 -0. 830

1383 C LEU A 187 25. .484 11 .840 -2. 199

1384 0 LEU A 187 24. .398 12 .274 -1. 807

1385 N VAL A 188 26, .649 12 .303 -1. 746

1386 CA VAL A 188 26, .683 13 .380 -0. 744

1387 CB VAL A 188 28, .102 13 .608 -0. 140

1388 CGI VAL A 188 28. .205 14 .960 0. 566

1389 CG2 VAL A 188 28. .444 12 .482 0. 831

1390 C VAL A 188 26, .003 14 .667 -1. 259

1391 0 VAL A 188 25, .019 15 .108 -0. 665

1392 N PRO A 189 26, .492 15 .251 -2. 375

1393 CA PRO A 189 25. .800 16 .415 -2. 928

1394 CB PRO A 189 26. .467 16 .589 -4. 293

1395 CG PRO A 189 27. .852 16 .120 -4. 068

1396 CD PRO A 189 27 , .720 14 .948 -3. 139

1397 C PRO A 189 24 , .284 16 .225 -3. 081

1398 0 PRO A 189 23, .522 17 .115 -2. 694

1399 N LEU A 190 23, .860 15 .078 -3. 617

1400 CA LEU A 190 22. .438 14 .776 -3. 798

1401 CB LEU A 190 22, .234 13 .430 -4. 505

1402 CG LEU A 190 20, .813 12 .855 -4. 614

1403 CD1 LEU A 190 19. .883 13 .745 -5. 443

1404 CD2 LEU A 190 20. ,838 11 .443 -5. 181

1405 C LEU A 190 21. , 677 14 .798 -2. 479

1406 0 LEU A 190 20. .612 15 .411 -2. 385

1407 N LEU A 191 22. .226 14 .138 -1. 465

1408 CA LEU A 191 21. .598 14 .130 -0. 148

1409 CB LEU A 191 22. ,275 13 .123 0. 783

1410 CG LEU A 191 22. .037 11 .636 0. 493

1411 CD1 LEU A 191 22. .702 10 .794 1. 577

1412 CD2 LEU A 191 20. .550 11 .287 0. 373

1413 C LEU A 191 21. .557 15 .513 0. 489

1414 0 LEU A 191 20. .557 15 .878 1. 110

1415 N LEU A 192 22. .631 16 .284 0. 323

1416 CA LEU A 192 22. .664 17 .665 0. 816

1417 CB LEU A 192 24. .069 18 .275 0. 692

1418 CG LEU A 192 25. .139 17 .782 1. 674

1419 CD1 LEU A 192 26. .538 18 .297 1. 293

1420 CD2 LEU A 192 24. .788 18 .154 3. 125

1421 C LEU A 192 21. .620 18 .555 0. 129

1422 0 LEU A 192 20. .943 19 .345 0. 787

1423 N MET A 193 21. .490 18 .420 -1. 189

1424 CA MET A 193 20. ,535 19 .225 -1. 945

1425 CB MET A 193 20. .815 19 .148 -3. 447

1426 CG MET A 193 22. .050 19 .907 -3. 903

1427 SD MET A 193 22. ,241 19 .981 -5. 701

1428 CE MET A 193 22. .305 18 .247 -6. 168

1429 C MET A 193 19. .096 18 .805 -1. 643

1430 0 MET A 193 18. .206 19 .652 -1. 551 1431 N LEU A 194 18..877 17.498 -1.496

1432 CA LEU A 194 17. , 573 16. 971 -1. 099

1433 CB LEU A 194 17. .556 15. 436 -1. 151

1434 CG LEU A 194 16. ,243 14. 707 -0. 816

1435 GDI LEU A 194 15. ,112 15. 082 -1. 772

1436 CD2 LEU A 194 16. .444 13. 197 -0. 800

1437 C LEU A 194 17. ,180 17. 482 0. 294

1438 0 LEU A 194 16. ,013 17. 809 0. 535

1439 N GLY A 195 18. .163 17. 563 1. 189

1440 CA GLY A 195 17. , 962 18. 088 2. 537

1441 C GLY A 195 17. .645 19. 571 2. 549

1442 0 GLY A 195 16. .879 20. 043 3. 394

1443 N VAL A 196 18. .243 20. 309 1. 615

1444 CA VAL A 196 17. .971 21. 736 1. 457

1445 CB VAL A 196 19, ,002 22. 414 0. 527

1446 CGI VAL A 196 18. .511 23. 782 0. 065

1447 CG2 VAL A 196 20. .340 22. 561 1. 238

1448 C VAL A 196 16. .545 21. 954 0. 947

1449 0 VAL A 196 15. .836 22. 836 1. 429

1450 N TYR A 197 16. ,131 21. 139 -0. 017

1451 CA TYR A 197 14. ,781 21. 201 -0. 550

1452 CB TYR A 197 14. , 627 20. 264 -1. 757

1453 CG TYR A 197 13. .191 19. 914 -2. 060

1454 CD1 TYR A 197 12. ,341 20. 836 -2. 663

1455 CE1 TYR A 197 11. .013 20. 521 -2. 925

1456 CZ TYR A 197 10. .529 19. 273 -2. 583

1457 OH TYR A 197 9. ,222 18. 956 -2. 838

1458 CE2 TYR A 197 11. ,350 18. 340 -1. 981

1459 CD2 TYR A 197 12. , 676 18. 665 -1. 720

1460 C TYR A 197 13. ,734 20. 891 0. 527

1461 0 TYR A 197 12. .708 21. 570 0. 617

1462 N LEU A 198 14. .002 19. 881 1. 350

1463 CA LEU A 198 13. .057 19. 473 2. 389

1464 CB LEU A 198 13. ,441 18. 109 2. 976

1465 CG LEU A 198 13. ,294 16. 908 2. 023

1466 CD1 LEU A 198 13. .986 15. 671 2. 570

1467 CD2 LEU A 198 11. .833 16. 597 1. 682

1468 C LEU A 198 12. .876 20. 529 3. 484

1469 0 LEU A 198 11. .810 20. 617 4. 098

1470 N ARG A 199 13. .906 21. 343 3. 701

1471 CA ARG A 199 13. .834 22. 435 4. 670

1472 CB ARG A 199 15. ,232 22. 826 5. 167

1473 CG ARG A 199 15. ,828 21. 838 6. 172

1474 CD ARG A 199 15. ,115 21. 922 7. 519

1475 NE ARG A 199 15. .045 20. 629 8. 199

1476 CZ ARG A 199 14. ,272 20. 372 9. 253

1477 NH1 ARG A 199 13. ,489 21. 318 9. 762

1478 NH2 ARG A 199 14. ,277 19. 163 9. 802

1479 C ARG A 199 13. , 091 23. 653 4. 127

1480 0 ARG A 199 12. ,416 24. 355 4. 885

1481 N ILE A 200 13. .220 23. 905 2. 824

1482 CA ILE A 200 12. , 473 24. 989 2. 177

1483 CB ILE A 200 13. , 083 25. 393 0. 805

1484 CGI ILE A 200 14. ,492 25. 966 1. 012

1485 CD1 ILE A 200 15. ,217 26. 376 -0. 261

1486 CG2 ILE A 200 12. .199 26. 421 0. 090

1487 C ILE A 200 10. .988 24. 615 2. 053

1488 0 ILE A 200 10. .107 25. 464 2. 226

1489 N PHE A 201 10. .727 23. 341 1. 774

1490 CA PHE A 201 9. .367 22. 825 1. 700 1491 CB PHE A 201 9.374 21.368 1.220

1492 CG PHE A 201 8. 005 20. 779 1. 036

1493 CD1 PHE A 201 7. 314 20. 952 -0. 161

1494 CE1 PHE A 201 6. 040 20. 407 -0. 337

1495 CZ PHE A 201 5. 449 19. 681 0. 689

1496 CE2 PHE A 201 6. 128 19. 497 1. 889

1497 CD2 PHE A 201 7. 404 20. 043 2. 056

1498 C PHE A 201 8. 645 22. 951 3. 047

1499 0 PHE A 201 7. 491 23. 387 3. 097

1500 N LEU A 202 9. 334 22. 569 4. 125

1501 CA LEU A 202 8. 793 22. 643 5. 482

1502 CB LEU A 202 9. 723 21. 936 6. 474

1503 CG LEU A 202 9. 398 20. 508 6. 940

1504 CD1 LEU A 202 9. 361 19. 490 5. 796

1505 CD2 LEU A 202 8. 090 20. 481 7. 728

1506 C LEU A 202 8. 519 24. 077 5. 939

1507 0 LEU A 202 7. 608 24. 317 6. 731

1508 N ALA A 203 9. 310 25. 020 5. 434

1509 CA ALA A 203 9. 081 26. 437 5. 681

1510 CB ALA A 203 10. 251 27. 265 5. 175

1511 C ALA A 203 7. 781 26. 886 5. 018

1512 0 ALA A 203 7. 013 27. 654 5. 603

1513 N ALA A 204 7. 545 26. 397 3. 800

1514 CA ALA A 204 6. 315 26. 686 3. 067

1515 CB ALA A 204 6. 426 26. 204 1. 625

1516 C ALA A 204 5. 095 26. 072 3. 757

1517 0 ALA A 204 4. 040 26. 696 3. 835

1518 N ARG A 205 5. 257 24. 855 4. 271

1519 CA ARG A 205 4. 193 24. 159 4. 992

1520 CB ARG A 205 4. 598 22. 709 5. 263

1521 CG ARG A 205 3. 424 21. 773 5. 450

1522 CD ARG A 205 3. 825 20. 327 5. 242

1523 NE ARG A 205 4. 442 19. 737 6. 430

1524 CZ ARG A 205 4. 743 18. 446 6. 558

1525 NHl ARG A 205 4. 483 17. 595 5. 571

1526 NH2 ARG A 205 5. 303 18. 000 7. 677

1527 C ARG A 205 3. 796 24. 868 6. 299

1528 0 ARG A 205 2. 621 24. 875 6. 670

1529 N ARG A 206 4. 772 25. 466 6. 981

1530 CA ARG A 206 4. 509 26. 211 8. 216

1531 CB ARG A 206 5. 796 26. 451 9. 013

1532 CG ARG A 206 6. 411 25. 190 9. 600

1533 CD ARG A 206 7. 571 25. 513 10. 531

1534 NE ARG A 206 8. 519 24. 403 10. 636

1535 CZ ARG A 206 8. 401 23. 374 11. 475

1536 NHl ARG A 206 9. 326 22. 422 11. 481

1537 NH2 ARG A 206 7. 365 23. 289 12. 305

1538 C ARG A 206 3. 799 27. 533 7. 940

1539 0 ARG A 206 2. 882 27. 908 8. 669

1540 N GLN A 207 4. 230 28. 231 6. 890

1541 CA GLN A 207 3. 582 29. 466 6. 449

1542 CB GLN A 207 4. 343 30. 093 5. 280

1543 CG GLN A 207 5. 666 30. 739 5. 647

1544 CD GLN A 207 5. 495 32. 039 6. 402

1545 0E1 GLN A 207 5. 849 32. 136 7. 579

1546 NE2 GLN A 207 4. 941 33. 045 5. 733

1547 C GLN A 207 2. 144 29. 194 6. 020

1548 0 GLN A 207 1. 242 29. 987 6. 295

1549 N LEU A 208 1. 949 28. 062 5. 348

1550 CA LEU A 208 0. 649 27. 668 4. 833 1551 CB LEU A 208 0..801 26.504 3.851

1552 CG LEU A 208 -0. .332 26 .247 2. 855

1553 CD1 LEU A 208 -0. .589 27 .456 1. 945

1554 CD2 LEU A 208 -0. .017 25 .008 2. 033

1555 C LEU A 208 -0. , 310 27 .295 5. 960

1556 0 LEU A 208 -1. .513 27 .540 5. 860

1557 N LYS A 209 0, .227 26 .716 7. 031

1558 CA LYS A 209 -0. .589 26 .339 8. 182

1559 CB LYS A 209 0, .147 25 .335 9. 073

1560 CG LYS A 209 -0, .797 24 .438 9. 851

1561 CD LYS A 209 -0, .064 23 .478 10. 760

1562 CE LYS A 209 -1. .050 22 .570 11. 478

1563 NZ LYS A 209 -0. .369 21 .695 12. 470

1564 C LYS A 209 -1. .057 27 .549 9. 000

1565 0 LYS A 209 -2, .089 27 .487 9. 670

1566 N GLN A 210 -0, .303 28 .643 8. 939

1567 CA GLN A 210 -0, .668 29 .879 9. 635

1568 CB GLN A 210 0. .540 30 .811 9. 765

1569 CG GLN A 210 1, .478 30 .444 10. 910

1570 CD GLN A 210 2, .839 31 .119 10. 815

1571 0E1 GLN A 210 3, .014 32 .114 10. 106

1572 NE2 GLN A 210 3. .814 30 .574 11. 537

1573 C GLN A 210 -1, .824 30 .601 8. 948

1574 0 GLN A 210 -2, .557 31 .360 9. 586

1575 N MET A 211 -1. .980 30 .351 7. 649

1576 CA MET A 211 -3, .065 30 .933 6. 859

1577 CB MET A 211 -2, .772 30 .794 5. 367

1578 CG MET A 211 -1, .563 31 .584 4. 913

1579 SD MET A 211 -1. .458 31 .723 3. 125

1580 CE MET A 211 0. .074 32 .632 2. 950

1581 C MET A 211 -4 , .431 30 .326 7. 202

1582 0 MET A 211 -5, .467 30 .801 6. 726

1583 N GLU A 212 -4 , .419 29 .284 8. 035

1584 CA GLU A 212 -5, .638 28 .692 8. 589

1585 CB GLU A 212 -5. .396 27 .230 8. 971

1586 CG GLU A 212 -5. .090 26 .337 7. 784

1587 CD GLU A 212 -4. .826 24 .894 8. 167

1588 OE1 GLU A 212 -4. .707 24 .597 9. 376

1589 OE2 GLU A 212 -4 , .733 24 .051 7. 250

1590 C GLU A 212 -6, .170 29 .488 9. 792

1591 0 GLU A 212 -6. .913 28 .959 10. 623

1592 N SER A 213 -5. .763 30 .754 9. 876

1593 CA SER A 213 -6. .289 31 .724 10. 837

1594 CB SER A 213 -5. .453 31 .747 12. 119

1595 OG SER A 213 -5. .395 30 .467 12. 720

1596 C SER A 213 -6. .261 33 .096 10. 170

1597 0 SER A 213 -5. .286 33 .439 9. 496

1598 N GLN A 214 -7. .331 33 .870 10. 343

1599 CA GLN A 214 -7. .443 35 .186 9. 700

1600 CB GLN A 214 -8, .864 35 .425 9. 157

1601 CG GLN A 214 -9. .993 35 .339 10. 192

1602 CD GLN A 214 -11. .380 35 .487 9. 574

1603 OE1 GLN A 214 -11. .680 34 .893 8. 537

1604 NE2 GLN A 214 -12. .233 36 .275 10. 217

1605 C GLN A 214 -7. .003 36 .327 10. 620

1606 O GLN A 214 -5. .940 36 .919 10. 425

1607 N SER A 223 4 , .086 37 .602 1. 905

1608 CA SER A 223 4. .758 37 .472 0. 614

1609 CB SER A 223 5. .244 38 .839 0. 125

1610 OG SER A 223 5. .776 38 .748 -1. 186 1611 C SER A 223 5.925 36.483 0.666

1612 0 SER A 223 6 .491 36. 124 -0 .372

1613 N THR A 224 6 .273 36. 049 1 .877

1614 CA THR A 224 7 .351 35. 082 2 .097

1615 CB THR A 224 7 .754 35. Oil 3 .591

1616 OG1 THR A 224 7 .883 36. 337 4 .115

1617 CG2 THR A 224 9 .080 34. 286 3 .770

1618 C THR A 224 6 .961 33. 690 1 .589

1619 0 THR A 224 7 .819 32. 917 1 .153

1620 N LEU A 225 5 .665 33. 385 1 .638

1621 CA LEU A 225 5 .144 32. 111 1 .145

1622 CB LEU A 225 3 .662 31. 952 1 .522

1623 CG LEU A 225 2 .913 30. 612 1 .395

1624 CD1 LEU A 225 3 .786 29. 398 1 .721

1625 CD2 LEU A 225 2 .278 30. 448 0 .026

1626 C LEU A 225 5 .360 31. 938 -0 .364

1627 0 LEU A 225 5 .696 30. 846 -0 .819

1628 N GLN A 226 5 .183 33. 014 -1 .129

1629 CA GLN A 226 5 .399 32. 971 -2 .579

1630 CB GLN A 226 4 .846 34. 228 -3 .258

1631 CG GLN A 226 4 .763 34. 128 -4 .786

1632 CD GLN A 226 3 .835 35. 161 -5 .414

1633 OE1 GLN A 226 3 .414 36. 123 -4 .764

1634 NE2 GLN A 226 3 .511 34. 963 -6 .693

1635 C GLN A 226 6 .873 32. 751 -2 .943

1636 0 GLN A 226 7 .179 32. 037 -3 .901

1637 N LYS A 227 7 .773 33. 367 -2 .177

1638 CA LYS A 227 9 .214 33. 187 -2 .367

1639 CB LYS A 227 10 .010 34. 146 -1 .477

1640 CG LYS A 227 9 .981 35. 597 -1 .937

1641 CD LYS A 227 10 .767 36. 498 -0 .993

1642 CE LYS A 227 10 .485 37. 971 -1 .264

1643 NZ LYS A 227 11 .074 38. 442 -2 .551

1644 C LYS A 227 9 .639 31. 747 -2 .093

1645 0 LYS A 227 10 .410 31. 165 -2 .859

1646 N GLU A 228 9 .117 31. 177 -1 .009

1647 CA GLU A 228 9 .454 29. 813 -0 .600

1648 CB GLU A 228 9 .015 29. 566 0 .839

1649 CG GLU A 228 9 .875 30. 292 1 .859

1650 CD GLU A 228 9 .282 30. 282 3 .253

1651 OE1 GLU A 228 9 .946 30. 796 4 .176

1652 OE2 GLU A 228 8 .156 29. 766 3 .432

1653 C GLU A 228 8 .888 28. 734 -1 .521

1654 0 GLU A 228 9 .560 27. 735 -1 .787

1655 N VAL A 229 7 .660 28. 932 -1 .999

1656 CA VAL A 229 7 .031 27. 988 -2 .928

1657 CB VAL A 229 5 .514 28. 284 -3 .129

1658 CGI VAL A 229 4 .928 27. 453 -4 .264

1659 CG2 VAL A 229 4 .745 28. 026 -1 .843

1660 C VAL A 229 7 .782 27. 990 -4 .263

1661 0 VAL A 229 8 .061 26. 929 -4 .827

1662 N HIS A 230 8 .119 29. 182 -4 .753

1663 CA HIS A 230 8 .942 29. 311 -5 .954

1664 CB HIS A 230 9 .084 30. 781 -6 .373

1665 CG HIS A 230 10 .022 30. 990 -7 .522

1666 ND1 HIS A 230 11 .330 31. 396 -7 .351

1667 CE1 HIS A 230 11 .920 31. 480 -8 .530

1668 NE2 HIS A 230 11 .044 31. 140 -9 .459

1669 CD2 HIS A 230 9 .850 30. 827 -8 .856

1670 C HIS A 230 10 .323 28. 656 -5 .761 1671 0 HIS A 230 10..799 27, .936 -6.644

1672 N ALA A 231 10. .945 28, .899 -4 .606

1673 CA ALA A 231 12. .250 28, .317 -4 .282

1674 CB ALA A 231 12. ,796 28, .911 -2 .989

1675 C ALA A 231 12. ,217 26, .786 -4 .195

1676 0 ALA A 231 13. ,090 26, .115 -4 .751

1677 N ALA A 232 11. ,208 26, .246 -3 .509

1678 CA ALA A 232 11. ,058 24 , .801 -3 .345

1679 CB ALA A 232 9. .902 24 , .486 -2 .416

1680 C ALA A 232 10. ,872 24 , .102 -4 .686

1681 0 ALA A 232 11. .388 23, .004 -4 .901

1682 N LYS A 233 10. , 146 24 , .756 -5 .586

1683 CA LYS A 233 9. , 951 24 , .248 -6 .938

1684 CB LYS A 233 8. , 900 25, .084 -7 .681

1685 CG LYS A 233 8. ,565 24 , .605 -9 .090

1686 CD LYS A 233 7. , 971 23, .203 -9 .108

1687 CE LYS A 233 7. ,747 22, .721 -10 .540

1688 NZ LYS A 233 7. ,165 21, .343 -10 .590

1689 C LYS A 233 11. ,283 24. .234 -7 .693

1690 0 LYS A 233 11. , 624 23. .238 -8 .342

1691 N SER A 234 12. ,034 25. .329 -7 .584

1692 CA SER A 234 13. ,354 25, .438 -8 .209

1693 CB SER A 234 13. , 950 26. .824 -7 .961

1694 OG SER A 234 13. ,318 27. .793 -8 .775

1695 C SER A 234 14. ,325 24. .352 -7 .735

1696 0 SER A 234 15. ,096 23. .814 -8 .523

1697 N LEU A 235 14. ,271 24. .028 -6 .449

1698 CA LEU A 235 15. ,140 23. .005 -5 .887

1699 CB LEU A 235 15. .289 23. .196 -4 .384

1700 CG LEU A 235 16. ,513 23. .997 -3 .940

1701 CD1 LEU A 235 16. ,524 25. .437 -4 .468

1702 CD2 LEU A 235 16. ,528 23. , 996 -2 .448

1703 C LEU A 235 14. , 686 21. .590 -6 .191

1704 0 LEU A 235 15. ,514 20. .677 -6 .262

1705 N ALA A 236 13. 380 21. ,400 -6 .351

1706 CA ALA A 236 12. 853 20. ,103 -6 .763

1707 CB ALA A 236 11. 331 20. ,076 -6 .659

1708 C ALA A 236 13. 313 19. .779 -8 .186

1709 0 ALA A 236 13. 652 18. .636 -8 .489

1710 N ILE A 237 13. 347 20. ,793 -9 .045

1711 CA ILE A 237 13. 807 20. , 620 -10 .423

1712 CB ILE A 237 13. 492 21. , 872 -11 .300

1713 CGI ILE A 237 11. 974 21. .991 -11 .503

1714 CD1 ILE A 237 11. 501 23. ,309 -12 .091

1715 CG2 ILE A 237 14. 182 21. .773 -12 .657

1716 C ILE A 237 15. 297 20. 222 -10 .466

1717 0 ILE A 237 15. 653 19. 207 -11 .070

1718 N ILE A 238 16. 142 21. 014 -9 .804

1719 CA ILE A 238 17. 574 20. 732 -9 .672

1720 CB ILE A 238 18. 254 21. 715 -8 .677

1721 CGI ILE A 238 18. 266 23. 141 -9 .243

1722 CD1 ILE A 238 18. 743 24. 208 -8 .259

1723 CG2 ILE A 238 19. 681 21. 257 -8 .347

1724 C ILE A 238 17. 852 19. 292 -9 .221

1725 0 ILE A 238 18. 626 18. 574 -9 .863

1726 N VAL A 239 17. 209 18. 889 -8 .123

1727 CA VAL A 239 17. 459 17. 599 -7 .474

1728 CB VAL A 239 16. 869 17. 567 -6, .030

1729 CGI VAL A 239 16. 808 16. 146 -5, .475

1730 CG2 VAL A 239 17. 678 18. 446 -5, .111 1731 C VAL A 239 16.915 16.440 -8..301

1732 0 VAL A 239 17. 600 15. 434 -8. .480

1733 N GLY A 240 15. 686 16. 590 -8. .798

1734 CA GLY A 240 15. 058 15. 582 -9. .648

1735 C GLY A 240 15. 881 15. 310 -10. .893

1736 0 GLY A 240 16. 164 14. 157 -11. .218

1737 N LEU A 241 16. 280 16. 379 -11. .580

1738 CA LEU A 241 17. 121 16. 260 -12. .770

1739 CB LEU A 241 17. 227 17. 600 -13. .500

1740 CG LEU A 241 15. 945 18. Oil -14, .229

1741 CD1 LEU A 241 16. 063 19. 446 -14. .692

1742 CD2 LEU A 241 15. 604 17. 067 -15. .401

1743 C LEU A 241 18. 511 15. 695 -12. .471

1744 0 LEU A 241 19. 006 14. 826 -13. .203

1745 N PHE A 242 19. 119 16. 185 -11. .390

1746 CA PHE A 242 20. 437 15. 729 -10. .949

1747 CB PHE A 242 20. 891 16. 528 -9. .721

1748 CG PHE A 242 22. 292 16. 219 -9. .272

1749 CD1 PHE A 242 23. 372 16. 939 -9. .775

1750 CE1 PHE A 242 24. 672 16. 652 -9. .364

1751 cz PHE A 242 24. 899 15. 635 -8. .435

1752 CE2 PHE A 242 23. 830 14. 912 -7. .927

1753 CD2 PHE A 242 22. 534 15. 208 -8. .340

1754 C PHE A 242 20. 457 14. 227 -10. .654

1755 0 PHE A 242 21. 426 13. 544 -10. .988

1756 N ALA A 243 19. 393 13. 717 -10. .042

1757 CA ALA A 243 19. 310 12. 291 -9. .724

1758 CB ALA A 243 18. 245 12. 033 -8. .664

1759 C ALA A 243 19. 047 11. 456 -10. .972

1760 0 ALA A 243 19. 596 10. 365 -11. .136

1761 N LEU A 244 18. 207 11. 985 -11. .851

1762 CA LEU A 244 17. 886 11. 341 -13. .113

1763 CB LEU A 244 16. 734 12. 088 -13. .785

1764 CG LEU A 244 15. 842 11. 336 -14. .765

1765 CD1 LEU A 244 14. 764 10. 560 -14. .015

1766 CD2 LEU A 244 15. 215 12. 327 -15. .732

1767 C LEU A 244 19. 101 11. 300 -14. .044

1768 0 LEU A 244 19. 283 10. 341 -14. .793

1769 N CYS A 245 19. 931 12. 336 -13. .993

1770 CA CYS A 245 21. 101 12. 417 -14. .871

1771 CB CYS A 245 21. 514 13. 873 -15. .098

1772 SG CYS A 245 20. 346 14. 820 -16. .101

1773 C CYS A 245 22. 300 11. 605 -14. .381

1774 0 CYS A 245 22. 981 10. 958 -15. .182

1775 N TRP A 246 22. 546 11. 631 -13. ,072

1776 CA TRP A 246 23. 773 11. 056 -12. ,506

1777 CB TRP A 246 24. 365 11. 999 -11. ,457

1778 CG TRP A 246 25. 067 13. 189 -12. ,058

1779 CD1 TRP A 246 24. 643 14. 486 -12. , 041

1780 NE1 TRP A 246 25. 550 15. 290 -12. , 692

1781 CE2 TRP A 246 26. 587 14. 518 -13. , 144

1782 CD2 TRP A 246 26. 316 13. 185 -12. ,768

1783 CE3 TRP A 246 27. 234 12. 185 -13. ,114

1784 CZ3 TRP A 246 28. 382 12. 546 -13. ,820

1785 CH2 TRP A 246 28. 617 13. 880 -14. ,182

1786 CZ2 TRP A 246 27. 738 14. 879 -13. ,847

1787 C TRP A 246 23. 683 9. 632 -11. ,945

1788 0 TRP A 246 24. 647 8. 874 -12. 050

1789 N LEU A 247 22. 547 9. 268 -11. 352

1790 CA LEU A 247 22. 419 7. 955 -10. ,704 1791 CB LEU A 247 21.166 7.865 -9.830

1792 CG LEU A 247 21 .067 8. 728 -8. 567

1793 CD1 LEU A 247 19 .683 8. 543 -7. 925

1794 CD2 LEU A 247 22 .185 8. 437 -7. 560

1795 C LEU A 247 22 .503 6. 740 -11. 641

1796 0 LEU A 247 23 .143 5. 752 -11. 277

1797 N PRO A 248 21 .870 6. 794 -12. 838

1798 CA PRO A 248 21 .927 5. 594 -13. 689

1799 CB PRO A 248 21 .193 6. 029 -14. 962

1800 CG PRO A 248 20 .228 7. 064 -14. 482

1801 CD PRO A 248 20 .982 7. 822 -13. 418

1802 C PRO A 248 23 .350 5. 109 -13. 999

1803 0 PRO A 248 23 .631 3. 922 -13. 844

1804 N LEU A 249 24 .238 6. 016 -14. 399

1805 CA LEU A 249 25 .629 5. 655 -14. 697

1806 CB LEU A 249 26 .403 6. 852 -15. 280

1807 CG LEU A 249 27 .871 6. 605 -15. 674

1808 CD1 LEU A 249 28 .028 5. 406 -16. 610

1809 CD2 LEU A 249 28 .505 7. 843 -16. 284

1810 C LEU A 249 26 .369 5. 085 -13. 482

1811 0 LEU A 249 27 . Ill 4. 101 -13. 597

1812 N HIS A 250 26 .169 5. 705 -12. 322

1813 CA HIS A 250 26 .857 5. 271 -11. 113

1814 CB HIS A 250 26 .940 6. 414 -10. 103

1815 CG HIS A 250 27 .891 7. 492 -10. 518

1816 ND1 HIS A 250 29 .198 7. 539 -10. 086

1817 CE1 HIS A 250 29 .805 8. 575 -10. 639

1818 NE2 HIS A 250 28 .943 9. 192 -11. 427

1819 CD2 HIS A 250 27 .741 8. 531 -11. 375

1820 C HIS A 250 26 .269 3. 994 -10. 522

1821 0 HIS A 250 27 .012 3. 143 -10. 029

1822 N ILE A 251 24 .950 3. 840 -10. 611

1823 CA ILE A 251 24 .309 2. 575 -10. 245

1824 CB ILE A 251 22 .766 2. 675 -10. 269

1825 CGI ILE A 251 22 .274 3. 530 -9. 094

1826 CD1 ILE A 251 20 .849 4. 058 -9. 270

1827 CG2 ILE A 251 22 .124 1. 289 -10. 205

1828 C ILE A 251 24 .817 1. 439 -11. 144

1829 0 ILE A 251 25 .114 0. 343 -10. 661

1830 N ILE A 252 24 .940 1. 707 -12. 444

1831 CA ILE A 252 25 .537 0. 738 -13. 362

1832 CB ILE A 252 25 .535 1. 238 -14. 829

1833 CGI ILE A 252 24 .123 1. 142 -15. 414

1834 CD1 ILE A 252 23 .891 2. 006 -16. 644

1835 CG2 ILE A 252 26 .514 0. 427 -15. 683

1836 C ILE A 252 26 .951 0. 362 -12. 910

1837 0 ILE A 252 27 .292 -0. 824 -12. 856

1838 N ASN A 253 27 .762 1. 370 -12. 580

1839 CA ASN A 253 29 .116 1. 148 -12. 056

1840 CB ASN A 253 29 .809 2. 487 -11. 748

1841 CG ASN A 253 30 .403 3. 159 -12. 997

1842 0D1 ASN A 253 30 .583 2. 529 -14. 036

1843 ND2 ASN A 253 30 .723 4. 439 -12. 882

1844 C ASN A 253 29 .139 0. 219 -10. 828

1845 0 ASN A 253 30 .021 -0. 629 -10. 707

1846 N CYS A 254 28 .153 0. 365 -9. 943

1847 CA CYS A 254 28 .016 -0. 502 -8. 767

1848 CB CYS A 254 26 .952 0. 035 -7. 816

1849 SG CYS A 254 27 .371 1. 646 -7. 139

1850 C CYS A 254 27 .700 -1. 945 -9. 134 1851 0 CYS A 254 28.258 -2..876 -8.540

1852 N PHE A 255 26. 809 -2. 126 -10 .109

1853 CA PHE A 255 26. 505 -3. 450 -10 .651

1854 CB PHE A 255 25. 431 -3. 358 -11 .745

1855 CG PHE A 255 25. 272 -4. 622 -12 .555

1856 CD1 PHE A 255 24. 474 -5. .666 -12 .092

1857 CE1 PHE A 255 24. 327 -6. 837 -12 .838

1858 CZ PHE A 255 24. 981 -6. 969 -14 .064

1859 CE2 PHE A 255 25. 783 -5. 933 -14 .535

1860 CD2 PHE A 255 25. 921 -4. 766 -13 .782

1861 C PHE A 255 27. 769 -4. 128 -11 .188

1862 0 PHE A 255 28. 037 -5. 290 -10 .873

1863 N THR A 256 28. 539 -3. 391 -11 .988

1864 CA THR A 256 29. 785 -3. 893 -12 .571

1865 CB THR A 256 30. 453 -2. 826 -13 .462

1866 0G1 THR A 256 29. 474 -2. 246 -14 .332

1867 CG2 THR A 256 31. 589 -3. 427 -14 .286

1868 C THR A 256 30. 777 -4. 317 -11 .490

1869 0 THR A 256 31. 437 -5. 351 -11 .614

1870 N PHE A 257 30. 875 -3. 514 -10 .432

1871 CA PHE A 257 31. 832 -3. 773 -9 .362

1872 CB PHE A 257 32. 082 -2. 507 -8 .543

1873 CG PHE A 257 33. 129 -2. 670 -7 .485

1874 CD1 PHE A 257 32. 771 -2. 724 -6 .138

1875 CE1 PHE A 257 33. 741 -2. 878 -5 .145

1876 CZ PHE A 257 35. 086 -2. 978 -5 .501

1877 CE2 PHE A 257 35. 455 -2. 925 -6 .846

1878 CD2 PHE A 257 34. 475 -2. 769 -7 .829

1879 C PHE A 257 31. 412 -4. 924 -8 .449

1880 0 PHE A 257 32. 246 -5. 748 -8 .067

1881 N PHE A 258 30. 128 -4. 976 -8 .104

1882 CA PHE A 258 29. 632 -5. 992 -7 .179

1883 CB PHE A 258 28. 495 -5. 445 -6 .309

1884 CG PHE A 258 28. 947 -4. 424 -5 .298

1885 CD1 PHE A 258 28. 437 -3. 129 -5 .321

1886 CE1 PHE A 258 28. 856 -2. 179 -4 .392

1887 CZ PHE A 258 29. 802 -2. 521 -3 .430

1888 CE2 PHE A 258 30. 324 -3. 811 -3 .402

1889 CD2 PHE A 258 29. 895 -4. 752 -4 .333

1890 C PHE A 258 29. 237 -7. 298 -7 .858

1891 0 PHE A 258 29. 049 -8. 308 -7 .180

1892 N CYS A 259 29. 120 -7. 289 -9 .184

1893 CA CYS A 259 28. 964 -8. 543 -9 .927

1894 CB CYS A 259 27. 573 -8. 681 -10 .561

1895 SG CYS A 259 27. 164 -10. 426 -10 .862

1896 C CYS A 259 30. 070 -8. 746 -10 .968

1897 0 CYS A 259 29. 875 -8. 453 -12 .153

1898 N PRO A 260 31. 239 -9. 251 -10 .525

1899 CA PRO A 260 32. 358 -9. 488 -11 .442

1900 CB PRO A 260 33. 530 -9. 774 -10 .498

1901 CG PRO A 260 32. 898 -10. 307 -9 .259

1902 CD PRO A 260 31. 578 -9. 608 -9 .133

1903 C PRO A 260 32. 118 -10. 675 -12 .379

1904 0 PRO A 260 32. 724 -10. 744 -13 .451

1905 N ASP A 261 31. 240 -11. 590 -11 .969

1906 CA ASP A 261 30. 908 -12. 780 -12. .757

1907 CB ASP A 261 30. 496 -13. 936 -11, .840

1908 CG ASP A 261 31. 654 -14. 471 -11, .008

1909 ODl ASP A 261 32. 829 -14. 250 -11, .381

1910 OD2 ASP A 261 31. 386 -15. 124 -9, .977 1911 C ASP A 261 29..823 -12.519 -13.804

1912 0 ASP A 261 29. .800 -13. 174 -14. 849

1913 N CYS A 262 28. .930 -11. 572 -13. 513

1914 CA CYS A 262 27. ,895 -11. 139 -14. 454

1915 CB CYS A 262 27. .065 -10. 004 -13. 856

1916 SG CYS A 262 25. .966 -10. 445 -12. 507

1917 C CYS A 262 28. .507 -10. 629 -15. 748

1918 0 CYS A 262 29. .554 -9. 977 -15. 732

1919 N SER A 263 27. .852 -10. 925 -16. 867

1920 CA SER A 263 28. .215 -10. 313 -18. 140

1921 CB SER A 263 27, .455 -10. 954 -19. 305

1922 OG SER A 263 26. .064 -11. 006 -19. 047

1923 C SER A 263 27. .932 -8. 816 -18. 054

1924 0 SER A 263 26. .961 -8. 387 -17. 417

1925 N HIS A 264 28. .796 -8. 028 -18. 681

1926 CA HIS A 264 28. .750 -6. 580 -18. 554

1927 CB HIS A 264 29. .925 -5. 959 -19. 302

1928 CG HIS A 264 30. .289 -4. 590 -18. 822

1929 ND1 HIS A 264 29. .566 -3. 467 -19. 157

1930 CE1 HIS A 264 30. .119 -2. 406 -18. 597

1931 NE2 HIS A 264 31. .181 -2. 800 -17. 918

1932 CD2 HIS A 264 31. .309 -4. 162 -18. 041

1933 C HIS A 264 27. .431 -6. 000 -19. 058

1934 0 HIS A 264 26. .838 -6. 520 -20. 002

1935 N ALA A 265 26. .972 -4. 932 -18. 408

1936 CA ALA A 265 25. .797 -4. 191 -18. 850

1937 CB ALA A 265 25. ,555 -2. 994 -17. 949

1938 C ALA A 265 25. .998 -3. 741 -20. 293

1939 0 ALA A 265 27. ,067 -3. 224 -20. 630

1940 N PRO A 266 24. ,976 -3. 949 -21. 152

1941 CA PRO A 266 25. .053 -3. 666 -22. 588

1942 CB PRO A 266 23. .586 -3. 734 -23. 047

1943 CG PRO A 266 22. .764 -3. 896 -21. 797

1944 CD PRO A 266 23. .667 -4. 520 -20. 796

1945 C PRO A 266 25. .654 -2. 308 -22. 933

1946 0 PRO A 266 25. .447 -1. 326 -22. 217

1947 N LEU A 267 26. .384 -2. 275 -24. 042

1948 CA LEU A 267 27. .069 -1. 080 -24. 528

1949 CB LEU A 267 27. .921 -1. 447 -25. 748

1950 CG LEU A 267 28. , 982 -0. 512 -26. 336

1951 CD1 LEU A 267 29. .822 0. 175 -25. 265

1952 CD2 LEU A 267 29. ,867 -1. 306 -27. 293

1953 C LEU A 267 26. ,133 0. 089 -24. 853

1954 0 LEU A 267 26. .533 1. 250 -24. 738

1955 N TRP A 268 24. ,898 -0. 210 -25. 258

1956 CA TRP A 268 23. .921 0. 842 -25. 572

1957 CB TRP A 268 22. ,726 0. 297 -26. 381

1958 CG TRP A 268 21. .801 -0. 599 -25. 596

1959 CD1 TRP A 268 21. .838 -1. 961 -25. 535

1960 NE1 TRP A 268 20. .844 -2. 428 -24. 708

1961 CE2 TRP A 268 20. ,138 -1. 363 -24. 214

1962 CD2 TRP A 268 20. ,710 -0. 190 -24. 755

1963 CE3 TRP A 268 20. ,166 1. 055 -24. 403

1964 CZ3 TRP A 268 19. ,077 1. 086 -23. 532

1965 CH2 TRP A 268 18. ,527 -0. 102 -23. 015

1966 CZ2 TRP A 268 19. 043 -1. 331 -23. 342

1967 C TRP A 268 23. ,448 1. 534 -24. 294

1968 0 TRP A 268 23. , 187 2. 738 -24. 290

1969 N LEU A 269 23. .338 0. 758 -23. 216

1970 CA LEU A 269 22. ,961 1. 293 -21. 911 1971 CB LEU A 269 22,.600 0,.156 -20,.954

1972 CG LEU A 269 22, .024 0, .485 -19, .574

1973 CD1 LEU A 269 20, .747 1. .316 -19. .660

1974 CD2 LEU A 269 21 , .780 -0. .808 -18, .794

1975 C LEU A 269 24 , .093 2 , .156 -21. .345

1976 0 LEU A 269 23, .837 3, .187 -20. .713

1977 N MET A 270 25, .336 1 , .740 -21. .595

1978 CA MET A 270 26. .501 2, .527 -21. .205

1979 CB MET A 270 27, .817 1, .811 -21. .556

1980 CG MET A 270 28, .114 0, .521 -20. .760

1981 SD MET A 270 28, .142 0, .648 -18. .939

1982 CE MET A 270 29, .359 1, .938 -18. .677

1983 C MET A 270 26, .444 3. .911 -21. .851

1984 0 MET A 270 26, .504 4 , .926 -21. .144

1985 N TYR A 271 26, .303 3, .941 -23. .181

1986 CA TYR A 271 26. .255 5, .192 -23. .951

1987 CB TYR A 271 26, .162 4, .920 -25. .455

1988 CG TYR A 271 27. .393 4 , .293 -26. .077

1989 CD1 TYR A 271 28. .678 4. .726 -25. .737

1990 CE1 TYR A 271 29. .810 4. .155 -26. .320

1991 CZ TYR A 271 29. .659 3. .147 -27. .261

1992 OH TYR A 271 30. .768 2. .574 -27. .843

1993 CE2 TYR A 271 28. .393 2. .710 -27. .621

1994 CD2 TYR A 271 27. .271 3. .286 -27. .032

1995 C TYR A 271 25. .101 6. .089 -23. .528

1996 0 TYR A 271 25. .265 7. .304 -23. .411

1997 N LEU A 272 23. .938 5. .484 -23. .299

1998 CA LEU A 272 22. .769 6. .201 -22. .782

1999 CB LEU A 272 21. .556 5. .265 -22. .704

2000 CG LEU A 272 20. .228 5. .803 -22. .163

2001 CD1 LEU A 272 19. .700 6. .974 -22. .983

2002 CD2 LEU A 272 19. .193 4. .683 -22. .080

2003 C LEU A 272 23. .040 6. .867 -21. .422

2004 0 LEU A 272 22. .710 8. .042 -21. .233

2005 N ALA A 273 23. .656 6. .123 -20. .499

2006 CA ALA A 273 23. .962 6. .624 -19. .149

2007 CB ALA A 273 24. .360 5. .473 -18. .226

2008 C ALA A 273 25. .032 7. .723 -19. .142

2009 0 ALA A 273 24. .897 8. .717 -18. ,426

2010 N ILE A 274 26. .081 7. .546 -19. , 946

2011 CA ILE A 274 27. ,125 8. ,565 -20. ,103

2012 CB ILE A 274 28. .271 8. .073 -21. ,034

2013 CGI ILE A 274 29. .047 6. , 920 -20. .379

2014 CD1 ILE A 274 29. ,831 6. ,043 -21. ,357

2015 CG2 ILE A 274 29. .209 9. ,223 -21. ,396

2016 C ILE A 274 26. ,514 9. ,864 -20. , 641

2017 0 ILE A 274 26. ,729 10. , 942 -20. ,085

2018 N VAL A 275 25. ,743 9. ,741 -21. ,720

2019 CA VAL A 275 25. ,027 10. ,867 -22. ,326

2020 CB VAL A 275 24. ,273 10. ,408 -23. 612

2021 CGI VAL A 275 23. ,077 11. 300 -23. 933

2022 CG2 VAL A 275 25. ,232 10. 342 -24. 790

2023 C VAL A 275 24. ,079 11. 534 -21. 319

2024 0 VAL A 275 24. 033 12. 765 -21. 211

2025 N LEU A 276 23. 341 10. 710 -20. 578

2026 CA LEU A 276 22. 424 11. 195 -19. 551

2027 CB LEU A 276 21. 700 10. 016 -18. 895

2028 CG LEU A 276 20. 235 10. 185 -18. 481

2029 CD1 LEU A 276 19. 338 10. 506 -19. 666

2030 CD2 LEU A 276 19. 750 8. 921 -17. 783 2031 C LEU A 276 23..154 12.047 -18..504

2032 0 LEU A 276 22. , 685 13. 119 -18. .138

2033 N SER A 277 24. .317 11. 580 -18. .050

2034 CA SER A 277 25. .119 12. 325 -17. .080

2035 CB SER A 277 26. ,296 11. 477 -16. .579

2036 OG SER A 277 27. .378 11. 446 -17. .489

2037 C SER A 277 25. .580 13. 679 -17. .625

2038 0 SER A 277 25. .624 14. 671 -16. .892

2039 N HIS A 278 25. ,893 13. 728 -18. .916

2040 CA HIS A 278 26. ,317 14. 981 -19. .543

2041 CB HIS A 278 27. .048 14. 716 -20. .855

2042 CG HIS A 278 28. .334 13. 972 -20. .678

2043 ND1 HIS A 278 28. .662 12. 861 -21. .422

2044 CE1 HIS A 278 29. .846 12. 415 -21. .043

2045 NE2 HIS A 278 30. .291 13. 188 -20. .069

2046 CD2 HIS A 278 29. .364 14. 169 -19. .821

2047 C HIS A 278 25. ,179 15. 983 -19. .733

2048 0 HIS A 278 25. .411 17. 196 -19. .715

2049 N THR A 279 23. .954 15. 479 -19. .880

2050 CA THR A 279 22. ,786 16. 347 -20. .026

2051 CB THR A 279 21. .557 15. 602 -20. .613

2052 OG1 THR A 279 20, .571 16. 557 -21. .014

2053 CG2 THR A 279 20. , 929 14. 673 -19. .606

2054 C THR A 279 22. , 422 17. 140 -18, .753

2055 0 THR A 279 21. .569 18. 032 -18, .794

2056 N ASN A 280 23. .075 16. 829 -17. .634

2057 CA ASN A 280 22. .950 17. 654 -16, .429

2058 CB ASN A 280 23. .618 16. 991 -15. .221

2059 CG ASN A 280 23, ,273 17. 688 -13. , 911

2060 ODl ASN A 280 22. .135 17. 639 -13. .458

2061 ND2 ASN A 280 24. .255 18. 345 -13. .305

2062 C ASN A 280 23, .535 19. 047 -16. .632

2063 0 ASN A 280 23, .107 20. 005 -15. .995

2064 N SER A 281 24 , .515 19. 148 -17. .526

2065 CA SER A 281 25, .202 20. 415 -17. .802

2066 CB SER A 281 26, .593 20. 155 -18. .390

2067 OG SER A 281 27, ,471 19. 647 -17. .400

2068 C SER A 281 24. .413 21. 368 -18. .705

2069 O SER A 281 24, ,777 22. 536 -18. .847

2070 N VAL A 282 23. ,345 20. 870 -19. .318

2071 CA VAL A 282 22. .464 21. 729 -20. .102

2072 CB VAL A 282 22. .090 21. 110 -21. .496

2073 CGI VAL A 282 21. .788 19. 633 -21. .389

2074 CG2 VAL A 282 20. .929 21. 862 -22. .152

2075 C VAL A 282 21. .244 22. 194 -19. .291

2076 O VAL A 282 20. , 820 23. 343 -19. .413

2077 N VAL A 283 20. ,705 21. 319 -18. .447

2078 CA VAL A 283 19. ,540 21. 679 -17. .630

2079 CB VAL A 283 18. .583 20. 478 -17. ,385

2080 CGI VAL A 283 18. ,038 19. 948 -18. , 717

2081 CG2 VAL A 283 19. ,268 19. 361 -16. .596

2082 C VAL A 283 19. , 906 22. 383 -16. .315

2083 O VAL A 283 19. , 168 23. 261 -15. .859

2084 N ASN A 284 21. ,043 22. 012 -15. .722

2085 CA ASN A 284 21. .510 22. 632 -14. .477

2086 CB ASN A 284 21. ,886 21. 567 -13. ,441

2087 CG ASN A 284 20. .681 20. 974 -12. ,730

2088 ODl ASN A 284 19. ,856 21. 695 -12. ,169

2089 ND2 ASN A 284 20. 596 19. 647 -12. ,720

2090 C ASN A 284 22. 712 23. 537 -14. ,723 2091 0 ASN A 284 23.725 23..072 -15.241

2092 N PRO A 285 22 .606 24. ,832 -14. 372

2093 CA PRO A 285 21 .420 25. .482 -13. 837

2094 CB PRO A 285 22 .010 26. .342 -12. 724

2095 CG PRO A 285 23 .335 26. ,798 -13. 308

2096 CD PRO A 285 23 .758 25. ,751 -14. 335

2097 C PRO A 285 20 .710 26. ,395 -14. 858

2098 0 PRO A 285 20 .345 27. ,526 -14. 511

2099 N PHE A 286 20 .521 25. , 937 -16. 096

2100 CA PHE A 286 19 .781 26. ,757 -17. 053

2101 CB PHE A 286 19, .782 26. ,181 -18. 477

2102 CG PHE A 286 19, .045 27. ,046 -19. 473

2103 CD1 PHE A 286 19, .638 28. ,198 -19. 991

2104 CE1 PHE A 286 18, .952 29. .012 -20. 907

2105 CZ PHE A 286 17, .658 28. ,675 -21. 304

2106 CE2 PHE A 286 17, .052 27. ,528 -20. 788

2107 CD2 PHE A 286 17, .745 26. ,725 -19. 874

2108 C PHE A 286 18, .354 26. , 964 -16. 555

2109 0 PHE A 286 17, .940 28. ,102 -16. 315

2110 N ILE A 287 17, .630 25. 858 -16. 379

2111 CA ILE A 287 16, .252 25. ,878 -15. 893

2112 CB ILE A 287 15, .688 24. 453 -15. 713

2113 CGI ILE A 287 15, .723 23. ,700 -17. 055

2114 CD1 ILE A 287 15, .218 22. 264 -17. 012

2115 CG2 ILE A 287 14, .267 24. 511 -15. 133

2116 C ILE A 287 16, .097 26. 688 -14. 602

2117 0 ILE A 287 15, .096 27. 385 -14. 420

2118 N TYR A 288 17. .092 26. 621 -13. 722

2119 CA TYR A 288 17, .054 27. 410 -12. 497

2120 CB TYR A 288 18. .229 27. 068 -11. 567

2121 CG TYR A 288 18. .329 27. 985 -10. 365

2122 CD1 TYR A 288 19. .243 29. 038 -10. 345

2123 CE1 TYR A 288 19. .332 29. 893 -9. 252

2124 CZ TYR A 288 18. .500 29. 700 -8. 171

2125 OH TYR A 288 18. .591 30. 551 -7. 094

2126 CE2 TYR A 288 17. .581 28. 663 -8. 165

2127 CD2 TYR A 288 17. .500 27. 813 -9. 261

2128 C TYR A 288 17. .014 28. 903 -12. 821

2129 0 TYR A 288 16. .122 29. 618 -12. 357

2130 N ALA A 289 17. ,976 29. 357 -13. 625

2131 CA ALA A 289 18. ,068 30. 758 -14. 043

2132 CB ALA A 289 19. .340 30. 990 -14. 856

2133 C ALA A 289 16. , 830 31. 181 -14. 835

2134 0 ALA A 289 16. ,246 32. 234 -14. 569

2135 N TYR A 290 16. ,429 30. 334 -15. 783

2136 CA TYR A 290 15. ,254 30. 569 -16. 623

2137 CB TYR A 290 15. ,064 29. 404 -17. 602

2138 CG TYR A 290 13. ,868 29. 547 -18. 527

2139 CD1 TYR A 290 13. ,886 30. 451 -19. 593

2140 CE1 TYR A 290 12. ,793 30. 584 -20. 441

2141 CZ TYR A 290 11. ,666 29. 806 -20. 230

2142 OH TYR A 290 10. ,585 29. 934 -21. 068

2143 CE2 TYR A 290 11. , 621 28. 899 -19. 184

2144 CD2 TYR A 290 12. 719 28. 774 -18. 339

2145 C TYR A 290 13. 965 30. 806 -15. 824

2146 0 TYR A 290 12. 988 31. 333 -16. 356

2147 N ARG A 291 13. 974 30. 419 -14. 551

2148 CA ARG A 291 12. 810 30. 560 -13. 685

2149 CB ARG A 291 12. 634 29. 312 -12. 823

2150 CG ARG A 291 12. 056 28. 111 -13. 559 2151 CD ARG A 291 11..987 26.908 -12.632

2152 NE ARG A 291 11. .520 27 .294 -11. 297

2153 CZ ARG A 291 10. .243 27 .344 -10. 924

2154 NHl ARG A 291 9. .278 27 .010 -11. 771

2155 NH2 ARG A 291 9. , 934 27 .715 -9. 692

2156 C ARG A 291 12. .878 31 .811 -12. 814

2157 0 ARG A 291 12. .027 32 .019 -11. 946

2158 N ILE A 292 13. .904 32 .628 -13. 031

2159 CA ILE A 292 13. .935 33 .981 -12. 481

2160 CB ILE A 292 15. .322 34 .362 -11. 880

2161 CGI ILE A 292 15. .670 33 .470 -10. 687

2162 CD1 ILE A 292 16. .730 32 .434 -10. 977

2163 CG2 ILE A 292 15. .351 35 .819 -11. 428

2164 C ILE A 292 13, .556 34 .923 -13. 623

2165 0 ILE A 292 14 , .209 34 .924 -14. 674

2166 N ARG A 293 12, .494 35 .704 -13. 424

2167 CA ARG A 293 11. .973 36 .587 -14. 477

2168 CB ARG A 293 10, .761 37 .400 -13. 990

2169 CG ARG A 293 10. .952 38 .168 -12. 682

2170 CD ARG A 293 9. .768 39 .083 -12. 384

2171 NE ARG A 293 9. .789 40 .299 -13. 198

2172 CZ ARG A 293 9. .124 40 .464 -14. 341

2173 NHl ARG A 293 8, .365 39 .491 -14. 833

2174 NH2 ARG A 293 9, .219 41 .613 -14. 998

2175 C ARG A 293 13, .049 37 .490 -15. 084

2176 0 ARG A 293 13, .172 37 .574 -16. 308

2177 N GLU A 294 13. .837 38 .129 -14. 217

2178 CA GLU A 294 14. .915 39 .031 -14. 620

2179 CB GLU A 294 15. .646 39 .558 -13. 383

2180 CG GLU A 294 16. .337 40 .908 -13. 578

2181 CD GLU A 294 15, .385 42 .098 -13. 495

2182 0E1 GLU A 294 14 , .398 42 .032 -12. 728

2183 OE2 GLU A 294 15, .636 43 .107 -14. 193

2184 C GLU A 294 15, .902 38 .374 -15. 593

2185 0 GLU A 294 16. .344 39 .005 -16. 555

2186 N PHE A 295 16. .239 37 .110 -15. 341

2187 CA PHE A 295 17. .043 36 .325 -16. 273

2188 CB PHE A 295 17. .474 34 .993 -15. 643

2189 CG PHE A 295 18, .842 35 .018 -15. 029

2190 CD1 PHE A 295 19, .978 34 .831 -15. 815

2191 CE1 PHE A 295 21. .253 34 .850 -15. 245

2192 CZ PHE A 295 21 , .394 35 .048 -13. 872

2193 CE2 PHE A 295 20. .267 35 .226 -13. 081

2194 CD2 PHE A 295 18. .999 35 .207 -13. 660

2195 C PHE A 295 16. .269 36 .038 -17. 553

2196 0 PHE A 295 16. .805 36 .177 -18. 655

2197 N ARG A 296 15. ,009 35 .634 -17. 393

2198 CA ARG A 296 14 , .166 35 .200 -18. 510

2199 CB ARG A 296 12, .853 34 .611 -17. 985

2200 CG ARG A 296 12, .038 33 .843 -19. 022

2201 CD ARG A 296 10. .801 33 .190 -18. 414

2202 NE ARG A 296 9. .929 34 .160 -17. 754

2203 CZ ARG A 296 9. .819 34 .304 -16. 436

2204 NHl ARG A 296 9. .002 35 .221 -15. 937

2205 NH2 ARG A 296 10. .520 33 .534 -15. 612

2206 C ARG A 296 13. .893 36 .325 -19. 510

2207 0 ARG A 296 13. .878 36 .092 -20. 717

2208 N GLN A 297 13. .690 37 .538 -19. 000

2209 CA GLN A 297 13. .461 38 .707 -19. 851

2210 CB GLN A 297 12. .903 39 .881 -19. 037 2211 CG GLN A 297 11.460 39.673 -18.572

2212 CD GLN A 297 10 .801 40 .937 -18 .037

2213 OE1 GLN A 297 11 .472 41 .901 -17 .654

2214 NE2 GLN A 297 9 .473 40 .933 -18 .003

2215 C GLN A 297 14 .726 39 .114 -20 .604

2216 0 GLN A 297 14 .666 39 .446 -21 .790

2217 N THR A 298 15, .864 39 .073 -19 .910

2218 CA THR A 298 17, .168 39 .381 -20 .501

2219 CB THR A 298 18, .273 39 .462 -19 .424

2220 OG1 THR A 298 17, .860 40 .351 -18 .376

2221 CG2 THR A 298 19, .586 39 .972 -20 .016

2222 C THR A 298 17, .538 38 .362 -21 .579

2223 0 THR A 298 18, .143 38 .717 -22 .592

2224 N PHE A 299 17, .156 37 .103 -21 .363

2225 CA PHE A 299 17 , .334 36 .053 -22 .368

2226 CB PHE A 299 16, .849 34 .690 -21 .853

2227 CG PHE A 299 17 , .762 34 .040 -20 .847

2228 CD1 PHE A 299 19, .113 34 .370 -20 .775

2229 CE1 PHE A 299 19, .948 33 .754 -19 .845

2230 CZ PHE A 299 19, .440 32 .785 -18 .988

2231 CE2 PHE A 299 18, .097 32 .436 -19 .059

2232 CD2 PHE A 299 17 , .268 33 .059 -19 .990

2233 C PHE A 299 16, .582 36 .382 -23 .652

2234 0 PHE A 299 17. .051 36 .067 -24 .748

2235 N ARG A 300 15, .409 37 .000 -23 .507

2236 CA ARG A 300 14. .576 37 .357 -24 .655

2237 CB ARG A 300 13, .163 37 .767 -24 .223

2238 CG ARG A 300 12. .330 36 .615 -23 .657

2239 CD ARG A 300 10. .827 36 .840 -23 .823

2240 NE ARG A 300 10. .353 38 .048 -23 .145

2241 CZ ARG A 300 9. .942 38 .097 -21 .880

2242 NH1 ARG A 300 9. .940 37 .005 -21 .124

2243 NH2 ARG A 300 9. .531 39 .249 -21 .368

2244 C ARG A 300 15. .235 38 .441 -25 .499

2245 0 ARG A 300 15. .327 38 .299 -26 .715

2246 N LYS A 301 15. .721 39 .497 -24 .842

2247 CA LYS A 301 16. .440 40, .591 -25 .512

2248 CB LYS A 301 16. .922 41, .639 -24 .503

2249 CG LYS A 301 15. .858 42, .147 -23 .536

2250 CD LYS A 301 16. .133 43, .584 -23 .076

2251 CE LYS A 301 17. .414 43, .718 -22 .255

2252 NZ LYS A 301 17. ,341 43, .019 -20 .937

2253 C LYS A 301 17. , 633 40, .112 -26 .347

2254 0 LYS A 301 17. ,933 40, .691 -27 .395

2255 N ILE A 302 18. ,308 39, .062 -25 .880

2256 CA ILE A 302 19. ,500 38, .538 -26 .554

2257 CB ILE A 302 20. ,426 37, .761 -25 .574

2258 CGI ILE A 302 20. ,970 38. .713 -24 .499

2259 CD1 ILE A 302 21. 452 38. .029 -23 .224

2260 CG2 ILE A 302 21. 576 37. .083 -26 .323

2261 C ILE A 302 19. 146 37. .695 -27 .787

2262 0 ILE A 302 19. ,701 37. .913 -28 .870

2263 N ILE A 303 18. 225 36. .745 -27 .620

2264 CA ILE A 303 17. 774 35. .896 -28 .729

2265 CB ILE A 303 16. 976 34. .652 -28 .238

2266 CGI ILE A 303 17. 714 33. .952 -27, .092

2267 CD1 ILE A 303 16. 844 33. , 022 -26, .258

2268 CG2 ILE A 303 16. 738 33. .663 -29, .385

2269 C ILE A 303 16. 950 36. .715 -29, .734

2270 0 ILE A 303 16. 977 36. .439 -30, .936 2271 N ARG A 304 16.234 37.723 -29.231

2272 CA ARG A 304 15. 486 38. 662 -30. 070

2273 CB ARG A 304 14. 725 39. 668 -29. 195

2274 CG ARG A 304 13. 602 40. 432 -29. 889

2275 CD ARG A 304 12. 586 40. 972 -28. 877

2276 NE ARG A 304 13. 138 42. 015 -28. 007

2277 CZ ARG A 304 12. 859 43. 317 -28. 098

2278 NHl ARG A 304 12. 022 43. 771 -29. 025

2279 NH2 ARG A 304 13. 418 44. 172 -27. 253

2280 C ARG A 304 16. 439 39. 384 -31. 023

2281 0 ARG A 304 16. 296 39. 286 -32. 246

2282 N SER A 305 17. 426 40. 076 -30. 449

2283 CA SER A 305 18. 437 40. 814 -31. 210

2284 CB SER A 305 19. 374 41. 570 -30. 267

2285 OG SER A 305 18. 658 42. 520 -29. 496

2286 C SER A 305 19. 248 39. 918 -32. 143

2287 0 SER A 305 19. 853 40. 400 -33. 100

2288 N HIS A 306 19. 259 38. 621 -31. 852

2289 CA HIS A 306 19. 916 37. 637 -32. 706

2290 CB HIS A 306 20. 242 36. 365 -31. 914

2291 CG HIS A 306 20. 721 35. 228 -32. 762

2292 ND1 HIS A 306 22. 020 35. 130 -33. 212

2293 CE1 HIS A 306 22. 152 34. 032 -33. 937

2294 NE2 HIS A 306 20. 984 33. 416 -33. 974

2295 CD2 HIS A 306 20. 072 34. 143 -33. 248

2296 C HIS A 306 19. 052 37. 308 -33. 924

2297 0 HIS A 306 19. 577 36. 967 -34. 984

2298 N VAL A 307 17. 733 37. 416 -33. 767

2299 CA VAL A 307 16. 791 37. 098 -34. 848

2300 CB VAL A 307 15. 483 36. 448 -34. 303

2301 CGI VAL A 307 14. 415 36. 337 -35. 386

2302 CG2 VAL A 307 15. 783 35. 067 -33. 725

2303 C VAL A 307 16. 515 38. 308 -35. 762

2304 0 VAL A 307 16. 469 38. 163 -36. 989

2305 N LEU A 308 16. 359 39. 494 -35. 171

2306 CA LEU A 308 16. 207 40. 728 -35. 954

2307 CB LEU A 308 15. 535 41. 849 -35. 135

2308 CG LEU A 308 16. 040 42. 335 -33. 769

2309 CD1 LEU A 308 17. 216 43. 307 -33. 877

2310 CD2 LEU A 308 14. 891 42. 992 -33. 019

2311 C LEU A 308 17. 525 41. 197 -36. 596

2312 0 LEU A 308 17. 636 42. 343 -37. 044

2313 N ARG A 309 18. 507 40. 297 -36. 644

2314 CA ARG A 309 19. 800 40. 558 -37. 275

2315 CB ARG A 309 20. 851 40. 943 -36. 228

2316 CG ARG A 309 20. 972 42. 442 -35. 998

2317 CD ARG A 309 22. 111 42. 770 -35. 043

2318 NE ARG A 309 21. 643 42. 956 -33. 669

2319 CZ ARG A 309 21. 450 44. 141 -33. 092

2320 NHl ARG A 309 21. 690 45. 265 -33. 759

2321 NH2 ARG A 309 21. 020 44. 203 -31. 838

2322 C ARG A 309 20. 294 39. 380 -38. 120

2323 0 ARG A 309 21. 302 39. 497 -38. 822

2324 N GLN A 310 19. 586 38. 254 -38. 048

2325 CA GLN A 310 19. 898 37. 079 -38. 868

2326 CB GLN A 310 20. 002 35. 816 -38. 007

2327 CG GLN A 310 21. 378 35. 587 -37. 382

2328 CD GLN A 310 22. 413 35. 069 -38. 375

2329 OE1 GLN A 310 22. 092 34. 312 -39. 294

2330 NE2 GLN A 310 23. 666 35. 472 -38. 183 2331 C GLN A 310 18.869 36.873 -39.974

2332 0 GLN A 310 19. 172 36. 282 -41. 013

2333 N GLN A 311 17. 655 37. 361 -39. 739

2334 CA GLN A 311 16. 572 37. 262 -40. 714

2335 CB GLN A 311 15. 212 37. 217 -40. 004

2336 CG GLN A 311 14. 996 36. 004 -39. 096

2337 CD GLN A 311 14. 618 34. 743 -39. 855

2338 OE1 GLN A 311 13. 465 34. 313 -39. 824

2339 NE2 GLN A 311 15. 588 34. 143 -40. 540

2340 C GLN A 311 16. 607 38. 413 -41. 722

2341 0 GLN A 311 15. 883 38. 390 -42. 719

2342 N GLU A 312 17. 463 39. 401 -41. 455

2343 CA GLU A 312 17. 554 40. 631 -42. 252

2344 CB GLU A 312 18. 599 41. 589 -41. 661

2345 CG GLU A 312 18. 383 43. 058 -42. 007

2346 CD GLU A 312 17. 197 43. 669 -41. 277

2347 OE1 GLU A 312 17. 419 44. 392 -40. 284

2348 OE2 GLU A 312 16. 044 43. 422 -41. 690

2349 C GLU A 312 17. 795 40. 411 -43. 755

2350 0 GLU A 312 17. 088 41. 008 -44. 569

2351 N PRO A 313 18. 784 39. 564 -44. 128

2352 CA PRO A 313 18. 974 39. 243 -45. 550

2353 CB PRO A 313 20. 041 38. 144 -45. 515

2354 CG PRO A 313 20. 847 38. 468 -44. 319

2355 CD PRO A 313 19. 868 38. 991 -43. 303

2356 C PRO A 313 17. 710 38. 734 -46. 252

2357 0 PRO A 313 17. 459 39. 098 -47. 402

2358 N PHE A 314 16. 928 37. 910 -45. 558

2359 CA PHE A 314 15. 715 37. 315 -46. 124

2360 CB PHE A 314 15. 440 35. 948 -45. 483

2361 CG PHE A 314 16. 478 34. 907 -45. 810

2362 CD1 PHE A 314 16. 352 34. 112 -46. 948

2363 CE1 PHE A 314 17. 314 33. 150 -47. 261

2364 CZ PHE A 314 18. 417 32. 977 -46. 429

2365 CE2 PHE A 314 18. 555 33. 767 -45. 290

2366 CD2 PHE A 314 17. 587 34. 726 -44. 986

2367 C PHE A 314 14. 495 38. 231 -46. 005

2368 0 PHE A 314 13. 556 38. 126 -46. 797

2369 N LYS A 315 14. 523 39. 122 -45. 015

2370 CA LYS A 315 13. 467 40. 114 -44. 806

2371 CB LYS A 315 13. 590 40. 730 -43. 408

2372 CG LYS A 315 12. 444 41. 647 -43. 009

2373 CD LYS A 315 12. 831 42. 516 -41. 820

2374 CE LYS A 315 11. 638 43. 261 -41. 254

2375 NZ LYS A 315 10. 675 42. 333 -40. 598

2376 C LYS A 315 13. 541 41. 205 -45. 877

2377 0 LYS A 315 12. 523 41. 579 -46. 470

2378 N ALA A 316 14. 756 41. 703 -46. no

2379 CA ALA A 316 15. 023 42. 696 -47. 146

2380 CB ALA A 316 16. 470 43. 156 -47. 079

2381 C ALA A 316 14. 695 42. 149 -48. 534

2382 0 ALA A 316 14. 109 42. 856 -49. 357

2383 N ALA A 317 15. 064 40. 891 -48. 780

2384 CA ALA A 317 14. 763 40. 222 -50. 046

2385 CB ALA A 317 15. 380 38. 825 -50. 083

2386 C ALA A 317 13. 259 40. 154 -50. 302

2387 0 ALA A 317 12. 793 40. 544 -51. 372

2388 N ALA A 318 12. 510 39. 679 -49. 308

2389 CA ALA A 318 11. 057 39. 535 -49. 413

2390 CB ALA A 318 10. 504 38. 814 -48. 189 2391 C ALA A 318 10.345 40..874 -49.613

2392 0 ALA A 318 9. 391 40. .959 -50. 387

2393 N ALA A 319 10. 813 41. .909 -48. 916

2394 CA ALA A 319 10. 240 43. ,252 -49. 031

2395 CB ALA A 319 10. 766 44 , .154 -47. 926

2396 C ALA A 319 10. 513 43. .868 -50. 405

2397 0 ALA A 319 9. 696 44. .629 -50. 926

2398 N GLU A 320 11. 665 43. .529 -50. 980

2399 CA GLU A 320 12. 036 43. .987 -52. 312

2400 CB GLU A 320 13. 545 43. .843 -52. 525

2401 CG GLU A 320 14. 110 44. .714 -53. 638

2402 CD GLU A 320 15. 582 44. .455 -53. 893

2403 OE1 GLU A 320 15. 922 43. .338 -54. 328

2404 OE2 GLU A 320 16. 402 45. .368 -53. 669

2405 C GLU A 320 11. 272 43. .207 -53. 380

2406 0 GLU A 320 10. 906 43. .765 -54. 417

2407 N ASN A 321 11. 038 41. .920 -53. 120

2408 CA ASN A 321 10. 271 41. .062 -54. 024

2409 CB ASN A 321 10. 362 39. .597 -53. 589

2410 CG ASN A 321 11. 759 39. .016 -53. 757

2411 ODl ASN A 321 12. 427 39. .243 -54. 768

2412 ND2 ASN A 321 12. 203 38. .250 -52. 764

2413 C ASN A 321 8. 806 41. .479 -54. 134

2414 0 ASN A 321 8. 183 41. .303 -55. 181

2415 N LEU A 322 8. 267 42. .030 -53. 048

2416 CA LEU A 322 6. 877 42. .480 -53. 006

2417 CB LEU A 322 6. 361 42. .515 -51. 562

2418 CG LEU A 322 4. 855 42. .710 -51. 352

2419 CD1 LEU A 322 4. 089 41. .434 -51. 672

2420 CD2 LEU A 322 4. 561 43. .170 -49. 931

2421 C LEU A 322 6. 722 43. .854 -53. 655

2422 0 LEU A 322 5. 681 44. .159 -54. 233

2423 N TYR A 323 7. 764 44. .673 -53. 547

2424 CA TYR A 323 7. 789 46. .011 -54. 133

2425 CB TYR A 323 9. 099 46. .707 -53. 751

2426 CG TYR A 323 9. 116 48. .214 -53. 920

2427 CD1 TYR A 323 8. 900 49. .060 -52. 832

2428 CE1 TYR A 323 8. 929 50. ,448 -52. 981

2429 cz TYR A 323 9. 186 50. , 997 -54. 232

2430 OH TYR A 323 9. 219 52. .362 -54. 394

2431 CE2 TYR A 323 9. 410 50. .180 -55. 323

2432 CD2 TYR A 323 9. 381 48. .796 -55. 163

2433 C TYR A 323 7. 615 45. .966 -55. 661

2434 0 TYR A 323 6. 771 46. , 674 -56. 210

2435 N PHE A 324 8. 402 45. ,125 -56. 334

2436 CA PHE A 324 8. 352 45. ,003 -57. 796

2437 CB PHE A 324 9. 763 44. ,819 -58. 386

2438 CG PHE A 324 10. 739 45. , 901 -57. 998

2439 CD1 PHE A 324 10. 506 47. ,231 -58. 341

2440 CE1 PHE A 324 11. 410 48. ,231 -57. 983

2441 CZ PHE A 324 12. 567 47. , 901 -57. 285

2442 CE2 PHE A 324 12. 812 46. ,575 -56. 945

2443 CD2 PHE A 324 11. 900 45. 585 -57. 304

2444 C PHE A 324 7. 434 43. 869 -58. 257

2445 0 PHE A 324 7. 515 43. 417 -59. 402

2446 N GLN A 325 6. 560 43. 421 -57. 362

2447 CA GLN A 325 5. 659 42. ,303 -57. 633

2448 CB GLN A 325 5. 043 41. ,814 -56. 322

2449 CG GLN A 325 4. 190 40. ,567 -56. 422

2450 CD GLN A 325 3. 408 40. ,321 -55. 149 2451 OE1 GLN A 325 2.454 41.039 -54.842

2452 NE2 GLN A 325 3. 812 39. 306 -54. 395

2453 C GLN A 325 4. 561 42. 705 -58. 614

2454 0 GLN A 325 3. 964 43. 774 -58. 482

2455 05' NEC A 400 31. 202 10. 037 -13. 637

2456 C5' NEC A 400 31. 745 11. 029 -14. 096

2457 N5 NEC A 400 32. 108 12. 077 -13. 353

2458 CIO NEC A 400 31. 620 12. 294 -12. 002

2459 Cll NEC A 400 32. 457 13. 305 -11. 239

2460 C4 ' NEC A 400 32. 061 11. 115 -15. 564

2461 04 ' NEC A 400 33. 121 10. 200 -15. 872

2462 CI' NEC A 400 32. 787 9. 371 -16. 991

2463 C2 ' NEC A 400 31. 525 9. 970 -17. 596

2464 02 ' NEC A 400 31. 889 10. 877 -18. 646

2465 C3' NEC A 400 30. 879 10. 717 -16. 447

2466 03' NEC A 400 30. 138 11. 850 -16. 917

2467 N9 NEC A 400 32. 470 7. 967 -16. 592

2468 C4 NEC A 400 32. 537 6. 907 -17. 403

2469 N3 NEC A 400 32. 902 6. 723 -18. 691

2470 C2 NEC A 400 32. 873 5. 499 -19. 251

2471 Nl NEC A 400 32. 501 4. 389 -18. 588

2472 C6 NEC A 400 32. 115 4. 423 -17. 289

2473 N6 NEC A 400 31. 753 3. 289 -16. 642

2474 C5 NEC A 400 32. 112 5. 740 -16. 610

2475 N7 NEC A 400 31. 824 6. 198 -15. 384

2476 C8 NEC A 400 32. 034 7. 542 -15. 384

2477 0 HOH C 1 33. 189 9. 056 -20. 360

2478 0 HOH C 2 32. 740 8. 423 -23. 192

2479 0 HOH C 3 30. 292 5. 291 -10. 082

2480 0 HOH C 4 23. 443 8. 394 -15. 587

2481 0 HOH C 5 18. 329 23. 751 -13. 272

2482 0 HOH C 6 14. 906 29. 893 -9. 727

2483 0 HOH C 7 30. 044 0. 105 -15. 574

2484 0 HOH C 8 35. 523 2. 006 -20. 601

2485 0 HOH C 9 26. 538 23. 501 -12. 664

2486 0 HOH C 10 32. 639 2. 195 -20. 429

2487 0 HOH C 11 35. 508 4. 325 -22. 396

2488 0 HOH C 12 31. 687 3. 559 -23. 050

2489 0 HOH C 13 15. 227 31. 370 -4. 820

2490 0 HOH C 14 22. 733 24. 857 -17. 433

2491 0 HOH C 15 33. 045 5. 972 -22. 523

2492 0 HOH C 16 45. 284 -1. 176 -20. 693

2493 0 HOH C 17 44. 376 -2. 531 -24. 635

2551 0 HOH C 18 22. 347 24. 848 -21. 304

2552 0 HOH C 19 15. 644 31. 623 -7. 650

2572 0 HOH C 20 33. 150 27. 372 -11. 365

2592 0 HOH c 21 42. 869 -1. 296 -22. 847

2593 0 HOH c 22 27. 455 16. 873 -16. 568

2594 0 HOH c 23 26. 665 18. 100 -14. 095

2595 0 HOH c 24 31. 864 5. 294 -29. 112

2596 0 HOH c 25 15. 687 24. 564 -11. 490

2597 0 HOH c 26 43. 428 -7. 975 -23. 534

2598 0 HOH c 27 25. 278 20. 905 -13. 526

2504 06 SOG A 501 43. 118 -3. 739 -2. 399

2505 C6 SOG A 501 41. 875 -4. 460 -2. 450

2506 C5 SOG A 501 40. 752 -3. 691 -1. 748

2507 C4 SOG A 501 39. 365 -4. 139 -2. 232

2508 04 SOG A 501 38. 484 -4. 351 -1. 124

2509 C3 SOG A 501 38. 715 -3. 103 -3. 141

2510 03 SOG A 501 39. 482 -2. 996 -4. 340 2511 C2 SOG A 501 38..638 -1..746 -2.451

2512 02 SOG A 501 37. ,413 -1. , 656 -1 .732

2513 05 SOG A 501 40. , 912 -2. .280 -1 .951

2514 CI SOG A 501 39. ,777 -1. , 560 -1 .457

2515 SI SOG A 501 40. ,076 0. , 167 -1 .183

2516 CI' SOG A 501 41. , 645 0. ,360 -1 .979

2517 C2' SOG A 501 41. , 996 1. ,840 -2 .070

2518 C3' SOG A 501 43. ,493 2. ,078 -1 .944

2519 C4 ' SOG A 501 43. ,888 3. ,474 -2 .408

2520 C5' SOG A 501 43. ,320 4. ,543 -1 .475

2521 C6' SOG A 501 43. ,797 5. , 978 -1 .740

2522 C7¹ SOG A 501 45. ,027 6. ,094 -2 .644

2523 C8 ' SOG A 501 45. ,413 7. ,545 -2 .858

2524 06 SOG A 502 49. ,235 -0. , 924 -9 .069

2525 C6 SOG A 502 48. ,295 0. ,105 -8 .733

2526 C5 SOG A 502 47. ,226 -0. , 424 -7 .769

2527 C4 SOG A 502 45. ,905 -0. ,734 -8 .469

2528 04 SOG A 502 46. ,087 -1. ,877 -9 .315

2529 C3 SOG A 502 44. 748 -1. ,017 -7 .511

2530 03 SOG A 502 43. ,534 -0. 752 -8 .223

2531 C2 SOG A 502 44. ,749 -0. 183 -6 .226

2532 02 SOG A 502 43. ,978 -0. 859 -5 .218

2533 05 SOG A 502 46. 982 0. 558 -6 .768

2534 CI SOG A 502 46. ,158 0. 087 -5 .700

2535 SI SOG A 502 46. 109 1. 309 -4 .419

2536 CI' SOG A 502 47. 337 2. 427 -5 .019

2537 C2 ' SOG A 502 47. 609 3. 540 -4 .024

2538 C3' SOG A 502 48. 996 4. 118 -4 .282

2539 C4 ' SOG A 502 48. 994 5. 629 -4 .093

2540 C5' SOG A 502 50. 378 6. 165 -3 .726

2541 C6' SOG A 502 50. 256 7. 291 -2 .702

2542 C7' SOG A 502 51. 216 8. 442 -2 .964

2543 C8 ' SOG A 502 50. 752 9. 659 -2 .195

2544 06 SOG A 503 35. 621 40. 787 -13 .298

2545 C6 SOG A 503 35. 319 41. 809 -12 .342

2546 C5 SOG A 503 34. 220 41. 340 -11 .391

2547 C4 SOG A 503 34. 174 42. 259 -10 .163

2548 04 SOG A 503 33. 757 43. 576 -10 .553

2549 C3 SOG A 503 33. 233 41. 724 -9 .090

2550 03 SOG A 503 33. 388 42. 474 -7 .878

2551 C2 SOG A 503 33. 544 40. 258 -8 .833

2552 02 SOG A 503 32. 651 39. 738 -7 .845

2553 05 SOG A 503 34. 438 39. 965 -11 .028

2554 CI SOG A 503 33. 439 39. 466 -10 .134

2555 SI SOG A 503 33. 639 37. 747 -9 .756

2556 CI' SOG A 503 34. 028 36. 952 -11 .280

2557 C2 ' SOG A 503 34. 279 35. 478 -10 .964

2558 C3' SOG A 503 34. 423 34. 632 -12 .222

2559 C4 ' SOG A 503 34. 679 33. 172 -11 .859

2560 C5' SOG A 503 35. 002 32. 357 -13 .105

2561 C6' SOG A 503 35. 710 31. 049 -12 .767

2562 C7' SOG A 503 36. 107 30. 322 -14 .052

2563 C8 ' SOG A 503 35. 818 28. 834 -14 .009

2564 06 SOG A 504 41. 476 26. 314 -8 .620

2565 C6 SOG A 504 40. 863 27. 349 -7 .835

2566 C5 SOG A 504 40. 593 28. 601 -8 .677

2567 C4 SOG A 504 39. 729 29. 610 -7 .922

2568 04 SOG A 504 40. 384 29. 970 -6, .701

2569 C3 SOG A 504 39. 492 30. 874 -8, .750

2570 03 SOG A 504 38. 423 31. 638 -8, .167 2571 C2 SOG A 504 39.152 30..586 -10, .215

2572 02 SOG A 504 39. 474 31. .752 -10, .987

2573 05 SOG A 504 39. ,931 28, .281 -9 .905

2574 CI SOG A 504 39. 903 29. .392 -10 .813

2575 SI SOG A 504 39. 117 28, .929 -12, .333

2576 CI' SOG A 504 40. 100 27, .534 -12, .771

2577 C2 ' SOG A 504 39. 737 27, .080 -14 , .176

2578 C3' SOG A 504 40. ,410 25 .753 -14 .497

2579 C4 ' SOG A 504 40. .308 25 .451 -15 .985

2580 C5' SOG A 504 41. ,094 24 .199 -16 .352

2581 C6' SOG A 504 41. ,034 23 .934 -17 .854

2582 C7 ' SOG A 504 42. ,384 23 .464 -18 .396

2583 C8' SOG A 504 42. .250 22 .718 -19 .710

2584 06 SOG A 505 40. .759 39 .323 -4 .091

2585 C6 SOG A 505 40. .177 40 .126 -5 .134

2586 C5 SOG A 505 40. .231 39 .405 -6 .486

2587 C4 SOG A 505 38. .923 39 .532 -7 .265

2588 04 SOG A 505 38, .557 40 .907 -7 .459

2589 C3 SOG A 505 39, .067 38 .820 -8 .606

2590 03 SOG A 505 37. .869 38 .954 -9 .384

2591 C2 SOG A 505 39, .376 37 .344 -8 .369

2592 02 SOG A 505 39, .660 36 .714 -9 .627

2593 05 SOG A 505 40, .500 38 .017 -6 .265

2594 CI SOG A 505 40, .562 37 .149 -7 .412

2595 SI SOG A 505 40, .600 35 .470 -6 .832

2596 CI' SOG A 505 39, .996 35 .698 -5 .183

2597 C2 ' SOG A 505 39, .611 34 .367 -4 .556

2598 C3' SOG A 505 39, .377 34 .512 -3 .052

2599 C4 ' SOG A 505 39 .491 33 .158 -2 .355

2600 C5' SOG A 505 40 .944 32 .722 -2 .168

2601 C6' SOG A 505 41 .474 31 .815 -3 .282

2602 C7 ' SOG A 505 42 .154 30 .579 -2 .707

2603 C8 ' SOG A 505 42 .706 29 .691 -3 .801

Table 1

Crystallographic table of statistics

outliers 1 0

Claims

1. A method of predicting a three dimensional structural representation of a target protein of unknown structure, or part thereof, comprising:

providing the coordinates of the human adenosine Aa_, receptor structure listed in Table (i), optionally varied by a root mean square deviation of residue backbone atoms of not more than .08 A, or selected coordinates thereof; and

predicting the three-dimensional structural representation of the target protein, or part thereof, by modelling the structural representation on all or the selected coordinates of the adenosine A_¾ receptor.

2. A method according to Claim 1 further comprising aligning the amino acid sequence of the target protein of unknown structure with the amino acid sequence of adenosine A_¾ receptor listed in Figure 14 (SEQ ID No: 2) to match homologous regions of the amino acid sequences prior to predicting the structural representation, and wherein modelling the structural representation comprises modelling the structural representation of the matched homologous regions of the target protein on the corresponding regions of the adenosine A_¾ receptor to obtain a three dimensional structural representation for the target protein that substantially preserves the structural representation of the matched homologous regions.

3. A method of predicting the three dimensional structural representation of a target protein of unknown structure, or part thereof, comprising:

providing the coordinates of the adenosine A_¾ receptor structure listed in Table (i), optionally varied by a root mean square deviation of residue backbone atoms of not more than 1.08 A, or selected coordinates thereof; and either (a) positioning the coordinates in the crystal unit cell of the protein so as to predict its structural representation, or (b) assigning NMR spectra peaks of the protein by manipulating the coordinates.

4. A method of predicting a three dimensional structural representation of a target protein of unknown structure, or part thereof, comprising:

providing the coordinates of the adenosine A_¾ receptor structure, optionally varied by a root mean square deviation of residue backbone atoms of not more than 1.08 A, or selected coordinates thereof;

providing an X-ray diffraction pattern of the target protein; and using the coordinates to predict at least part of the structure coordinates of the target protein.

5. A method according to any of Claims 1 - 4, wherein the target protein is a GPCR, such as any of an adrenergic receptor, an adenosine receptor, a muscarinic receptor, a purinergic receptor, a dopaminergic receptor or a chemokine receptor.

6. A method for selecting or designing one or more binding partners of adenosine A2A receptor comprising using molecular modelling means to select or design one or more binding partners of adenosine A_¾ receptor, wherein the three-dimensional structural representation of at least part of adenosine A2A receptor , as defined by the coordinates of the human adenosine A2A receptor listed in Table (i), optionally varied by a root mean square deviation of residue backbone atoms of not more than 1.08 A, or selected coordinates thereof, is compared with a three-dimensional structural representation of one or more candidate binding partners, and one or more binding partners that are predicted to interact with adenosine A2A receptor are selected.

7. A method for producing a binding partner of adenosine A2A receptor comprising: identifying a binding partner according to the method of Claim 6, and

synthesising the binding partner. 8. A binding partner produced by the method of Claim 7 .

9. A method of predicting the three dimensional structure of a binding partner of unknown structure, or part thereof, which binds to adenosine A2A receptor , comprising:

providing the coordinates of the human adenosine A_¾ receptor structure, listed in Table (i), optionally varied by a root mean square deviation of residue backbone atoms of not more than 1.08 A, or selected coordinates thereof;

providing an X-ray diffraction pattern of adenosine A2A receptor complexed with the binding partner; and

using said coordinates to predict at least part of the structure coordinates of the binding partner.

10. A method according to Claim 9, wherein the X-ray diffraction pattern is obtained from a crystal formed either by (a) soaking a crystal of adenosine A_M receptor with the binding partner to form a complex, or (b) mixing adenosine A_∑A receptor with the binding partner and crystallising an adenosine A2A receptor -binding partner complex.

11. A method for producing a medicament, pharmaceutical composition or drug, the process comprising: (a) providing a binding partner according to Claim 8 and (b) preparing a medicament, pharmaceutical composition or drug containing the binding partner. 12. A method of obtaining a three dimensional structural representation of a crystal of a adenosine A2A receptor , which method comprises providing the coordinates of the human adenosine A¾ receptor structure, listed in Table (i), optionally varied by a root mean square deviation of residue backbone atoms of not more than 1.08 A, or selected coordinates thereof, and generating a three-dimensional structural representation of said coordinates. 3. A method for assessing the activation state of a structure for adenosine A2A receptor, comprising: providing the coordinates of the adenosine A2A receptor structure, listed in Table (i), optionally varied by a root mean square deviation of residue backbone atoms of not more than 1.08 A, or selected coordinates thereof; performing a statistical and/or a topological analysis on the coordinates; and comparing the results of the analysis with the results of an analysis of coordinates of proteins of known activation states.

14. A computer system, intended to generate three dimensional structural representations of adenosine A_¾ receptor , adenosine A¾ receptor homologues or analogues, complexes of adenosine A¾ receptor with binding partners, or complexes of adenosine A_¾ receptor homologues or analogues with binding partners, or, to analyse or optimise binding of binding partners to said adenosine A¾ receptor or homologues or analogues, or complexes thereof, the system containing computer-readable data comprising one or more of:

(a) the coordinates of the human adenosine A₂A receptor structure, listed in Table (i), optionally varied by a root mean square deviation of residue backbone atoms of not more than 1.08 A, or selected coordinates thereof;

(c) the coordinates of a binding partner generated by interpreting X-ray crystallographic data or N R data by reference to the coordinates of the adenosine A¾ receptor structure, listed in Table (i), optionally varied by a root mean square deviation of residue backbone atoms of not more than 1.08 A, or selected coordinates thereof, and

(d) structure factor data derivable from the coordinates of (a), (b) or (c).

15. A computer-readable storage medium, comprising a data storage material encoded with computer readable data, wherein the data comprises one or more of

(a) the coordinates of the human adenosine A_¾ receptor structure, listed in Table (i), optionally varied by a root mean square deviation of residue backbone atoms of not more than 1.08 A, or selected coordinates thereof;

(b) the coordinates of a target adenosine A_¾ receptor homologue or analogue generated by homology modelling of the target based on the data in (a);

(c) the coordinates of a binding partner generated by interpreting X-ray crystallographic data or NMR data by reference to the coordinates of the human adenosine A2A receptor structure, listed in Table (i), optionally varied by a root mean square deviation of residue backbone atoms of not more than 1.08 A, or selected coordinates thereof, and

(d) structure factor data derivable from the coordinates of (a), (b) or (c).

16. A computer-readable storage medium comprising a data storage material encoded with a first set of computer-readable data comprising a Fourier transform of at least a portion of the structural coordinates of human adenosine A2A receptor listed in Table (i), optionally varied by a root mean square deviation of residue backbone atoms of not more than 1.08 A, or selected coordinates thereof; which data, 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 structure coordinates corresponding to the second set of machine readable data.

17. A method of producing a protein with a binding region that has substrate specificity substantially identical to that of adenosine A_¾ receptor , the method comprising:

a) aligning the amino acid sequence of a target protein with the amino acid sequence of an adenosine A¾ receptor;

b) identifying the amino acid residues in the target protein that contribute to the adenosine binding site of the A_2A receptor and correspond to any one or more of the following positions according to the numbering of the adenosine A2A receptor as set out in Figure 13: Ile66, Ala63, lie 274, Val84, His278, Ser277, Leu249, Trp246, Thr88, Asn181 , His250, Met177, Asn253, Met 270, Glu169 and Phe168; or identifying the amino acid residues in the target protein that contribute to the NECA binding site of the A2A receptorand correspond to any one or more of the following positions according to the numbering of the adenosine AZA receptor as set out in Figure 13 (SEQ ID No: 1): Ile66, Ala63, lie 274, Val84, His278, Ser277, Leu249, Trp246, Thr88, Asn181 , His250, et177, Asn253, Met 270, Glu169, Phe168, Leu85 and Tyr9;

c) making one or more mutations in the amino acid sequence of the target protein to replace one or more identified amino acid residues with the corresponding residue in the adenosine A2A receptor.

18. A mutant adenosine A2A receptor which, when compared to the corresponding wild- type adenosine receptor, has a different amino acid at a position which corresponds to any one or more of the following positions according to the numbering of the human adenosine AZA receptor as set out in Figure 13 (SEQ ID No: 1): Ile66, Ala63, lie 274, Val84, His278, Ser277, Leu249, Trp246, Thr88, Asn181 , His250, Met177, Asn253, Met 270, Glu169 and Phe168.

19. A mutant adenosine A_¾ receptor which, when compared to the corresponding wild- type adenosine receptor, has a different amino acid at a position which corresponds to any one or more of the following positions according to the numbering of the human adenosine A2A receptor as set out in Figure 13 (SEQ ID No: 1): Ile66, Ala63, lie 274, Val84, His278, Ser277, Leu249, Trp246, Thr88, Asn181 , His250, Met177, Asn253, Met 270, Glu169, Phe168, Leu85 and Tyr9.

20. A method of making an adenosine A_¾ receptor crystal comprising:

providing purified adenosine A2A receptor ; and

crystallising the adenosine A^ receptor either by using the sitting drop or hanging drop vapour diffusion technique, using a precipitant solution comprising 0.05 M ADA NaOH, pH 6.4, 23.6% PEG 400, 4% v/v 2- propanol.

21. A method according to Claim19, wherein the precipitant solution comprises 0.05 M TrisHCI, pH 7.6, 9.6% PEG 200, 22.9%. PEG 300. Crystals were cryo-protected by soaking in 0.05 M TrisHCI, pH 7.5, 15% PEG 200, 30% PEG 300.

22. A crystal of adenosine A_2A receptor having the structure defined by the coordinates of the human adenosine A2A receptor structure listed in Table (i) optionally varied by a root mean square deviation of residue backbone atoms of not more than 1.08 A, or selected coordinates thereof.

23. A crystal according to Claim 22, which has C2 symmetry with unit cell dimensions a=76.9 (± 10) A, b=99.6 (± 10) A, c=79.7 (± 10) A, wherein a= 90 β = 93.3 (± 10)° and γ = 90 for NECA; a=76.5 (± 15) A, b=98.9 (± 15) A, c=79.5 (+ 15) A, wherein a= 90 )°, β = 93.5 (± 10)° and y = 90 (± 10)°.

24. A co-crystal of adenosine A2A receptor having the structure defined by the coordinates of the adenosine A2A receptor structure listed in Table (i) optionally varied by a root mean square deviation of residue backbone atoms of not more than 1.08 A, or selected coordinates thereof, and a binding partner.

25. A crystal according to any of Claims 22-24 having a resolution of 3.0 A or better.

26. A method of predicting a three dimensional structural representation of an active state of a target protein, or part thereof, comprising:

providing the coordinates of the human adenosine A_¾ receptor structure listed in Table (i), optionally varied by a root mean square deviation of residue backbone atoms of not more than 1.08 A, or selected coordinates thereof; and

predicting the three-dimensional structural representation of an active state or partially active state of the target protein, or part thereof, by modelling a structural representation of the target protein in a non-active state on all or the selected coordinates of the adenosine A2A receptor.

27. A method of predicting a three dimensional structural representation of an inactive state of a target protein, or part thereof, comprising:

predicting the three-dimensional structural representation of the inactive state of the target protein, or part thereof, by modelling a structural representation of the target protein in an active state on all or the selected coordinates of the adenosine A_¾ receptor.