Crystallized P38 complexes

This application claims priority from U.S. Provisional Applications Ser. No. 60/112,354 filed Dec. 16, 1998, and U.S. Provisional Application Ser. No. 60/163,373 filed Nov. 3, 1999.

This invention relates to certain crystallized kinase protein-ligand complexes, particularly complexes of crystallized P38 protein, and more particularly complexes of P38γ protein. This invention also relates to crystallizable compositions from which the protein-ligand complexes may be obtained. This invention also relates to computational methods of using structure coordinates of the protein complex to screen for and design compounds that interact with the protein, particularly P38 protein or homologues thereof.

Mammalian cells respond to extracellular stimuli by activating signaling cascades that are mediated by members of the mitogen-activated protein (MAP) kinase family. Mammalian mitogen-activated protein (MAP) kinases are proline-directed serine/threonine kinases that facilitate signal translocation in cells [Davis, Mol. Reprod. Dev. 42, 459-467 (1995); Cobb et al., J. Biol. Chem. 270, 14843-14846 (1995); Marshall, Cell 80, 179-185 (1995)]. MAP kinases include the extracellular-signal regulated kinases (ERKs), the c-Jun NH 2 -terminal kinases (JNKs) and the P38 kinases, which have similar sequences and three-dimensional structures [Taylor & lipopolysaccharides (LPS), UV, anisomycin, or osmotic shock, and by cytokines, such as interleukin-1 (IL-1) and tissue necrosis factor (TNF). Inhibition of P38α kinase leads to a blockade on the production of both IL-1 and TNF. IL-1 and TNF stimulate the production of other proinflammatory cytokines such as IL-6 and IL-8 and have been implicated in acute and chronic inflammatory diseases and in post-menopausal osteoporosis [Kimble et al., Endocrinol., 136, 3054-61 (1995)].

Based upon this finding it is believed that P38α, along with other MAPKs, has a role in mediating cellular response to inflammatory stimuli, such as leukocyte accumulation, macrophage/monocyte activation, tissue resorption, fever, acute phase responses and neutrophilia. In addition, the MAPKs, such as P38α, have been implicated in cancer, thrombin-induced platelet aggregation, immunodeficiency disorders, autoimmune diseases, cell death, allergies, osteoporosis and neurodegenerative disorders. Inhibitors of P38α also appear to be involved in pain management through inhibition of prostaglandin endoperoxide synthase-2 induction. Other diseases associated with Il-1, IL-6, IL-8 or TNF overproduction are set forth in WO 96/21654. P38γ MAP kinase (also known as ERK6 and stress activated protein kinase-3 or SAPK3) is a newly discovered member of the MAP kinase family. However, unlike the other P38 family members which are expressed in many tissues, P38γ is expressed at highest levels in skeletal muscle [Li et al., Biochem Biophys Res Commun 228, 334-340 (1996); Enslen et al., J Biol Chem 273, 1741-1748 (1998); Raingeaud et al., J. Biol. Chem. 270, 7420-7426 (1995)]. Thus P38γ may have a unique function related to muscle morphogenesis, and it may be a potential target for treating degenerative diseases occurring in muscle tissue.

Compounds that selectively inhibit P38γ and not P38α would be highly desirable. It would be useful to have new treatments for muscle degenerative diseases using compounds that do not suppress the inflammatory response or other functions of P38α. However, the design of inhibitors that are selective for any particular MAP kinase, such as P38γ, is challenging due to the structural similarity of the MAP kinases. Therefore, it would be advantageous to have a detailed understanding of the structures of the various MAP kinases in order to exploit any subtle differences that may exist among them.

A general approach to designing inhibitors that are selective for an enzyme target is to determine how a putative inhibitor interacts with the three dimensional structure of the enzyme. For this reason it is useful to obtain the enzyme protein in crystal form and perform X-ray diffraction techniques to determine its three dimensional structure coordinates. If the enzyme is crystallized as a complex with a ligand, one can determine both the shape of the enzyme binding pocket when bound to the ligand, as well as the amino acid residues that are capable of close contact with the ligand. By knowing the shape and amino acid residues in the binding pocket, one may design new ligands that will interact favorably with the enzyme. With such structural information, available computational methods may be used to predict how strong the ligand binding interaction will be. Such methods thus enable the design of inhibitors that bind strongly, as well as selectively to the target enzyme.

Crystal structures are known for some of the MAP kinases; for example, unphosphorylated JNK3, unphosphorylated P38α, and ERK2 in both phosphorylated and unphosphorylated forms. Phosphorylated ERK2 is reported to exist as a dimer in both solution and as a crystal. The unphosphorylated forms of JNK3, ERK2 and P38α, on the other hand, are reported to be monomeric. [Tong et al., Nat Struct Biol 4, 311-316 (1997); Wilson and Su, Chem Biol 4, 423-431 (1997); Xie et al., Structure 6, 983-991 (1998); Zhang et al., Nature 367, 704-711 (1994); Canagarajah et al., Cell 90, 859-869 (1997); Wilson and Su, J Biol Chem 271, 27696-27700 (1996)].

The crystal structure reported for P38α is based on unphosphorylated protein. However, it is the phosphorylated or activated form of the enzyme that is able to phosphorylate its substrate enzyme. In order to disrupt the phosphorylation of the substrate, and produce the desired clinical effect, a small molecule inhibitor would likely act by blocking a phosphorylated form of P38. Thus, the most suitable target for drug design is the active or phosphorylated form. While the structure of the unphosphorylated enzyme is often used for drug design purposes, there is an inherent uncertainty as to whether the phosphorylated and unphosphorylated forms would bind a designed inhibitor with equal affinity.

A class of pyridinylimidazole compounds are known to inhibit P38α MAP kinase [Lee et al., Nature 372, 739-746 (1994)]. These inhibitors have been shown to bind in the ATP binding site of P38α [Young et al., J Biol Chem 272, 12116-12121 (1997); Tong et al., Nat Struct Biol 4, 311-316 (1997); Wilson et al., Chem Biol 4, 423-431 (1997)]. However, the pyridinylimidazoles reportedly do not inhibit the activity of ERK2, JNK3, or P38γ. This observed selectivity is interesting because the amino acid sequence in the ATP binding site of the various kinases are known to be highly conserved [Fox et al., Protein Science 7, 2249-2255 (1998); Xie et al., supra; Wilson and Su, supra; Enslen et al., J Biol Chem 273, 1741-1748 (1998)].

As there is a need for compounds that selectively inhibit a particular MAP kinase, it would be desirable to have improved methods that facilitate the design of such compounds. For this purpose, knowledge of the three dimensional structure coordinates of an activated P38 protein would be useful. Such information would aid in identifying and designing potential inhibitors of particular P38 proteins which, in turn, are expected to have therapeutic utility.

This invention provides certain crystallized, protein kinase-ligand complexes, in particular P38-ligand complexes, and their structure coordinates. The structure coordinates are based on the structure of a phosphorylated P38γ-ligand complex that has now been solved and which reveals new structural information useful for understanding the activated states of other, related kinase proteins as described herein. The key structural features of the proteins, particularly the shape of the substrate binding site, are useful in methods for designing or identifying selective inhibitors of the protein kinases, particularly P38, and in solving the structures of other proteins with similar features.

The invention also provides a computer which which is programmed with the structure coordinates of the activated P38 binding site. Such a computer, appropriately programmed and attached to the necessary viewing device, is capable of displaying a three-dimensional graphical representation of a molecule or molecular complex comprising such binding sites or similarly shaped homologous binding pockets.

The invention also provides a method for determining at least a portion of the three-dimensional structure of other molecules or molecular complexes which contain at least some features that are structurally similar to P38γ, particularly P38α, P38β, P38δ and other P38 isoforms. This is achieved by using at least some of the structural coordinates obtained for a phosphorylated P38 complex.

FIGS. 1 . 1 - 1 . 98 lists the atomic structure coordinates for phosphorylated P38γ in complex with MgAMP-PNP as derived by X-ray diffraction from a crystal of that complex. The following abbreviations are used in FIGS. 1 . 1 - 1 . 98 :

“X, Y, Z” crystallographically define the atomic position of the element measured.

“B” is a thermal factor that measures movement of the atom around its atomic center.

“Occ” is an occupancy factor that refers to the fraction of the molecules in which each atom occupies the position specified by the coordinates. A value of “1” indicates that each atom has the same conformation, i.e., the same position, in all molecules of the crystal.

FIG. 2 is an overview of the phosphorylated P38γ.

FIG. 3 is a superimposition of unphosphorylated P38γ and phosphorylated P38γ.

FIG. 4 is a detailed stereo view of the activation loop.

FIG. 5 is a stereo view of the AMP-PNP bound in the active site.

FIG. 6 is a comparison of the active sites of activated P38γ with P38α (a) and cAPK or cyclic AMP dependent protein kinase(b).

FIG. 7 is a comparison of activated phosphorylation loops from P38γ (dark orange), ERK2 (dark blue), and cAPK (red).

FIG. 8 shows a diagram of a system used to carry out the instructions encoded by the storage medium of FIGS. 9 and 10.

FIG. 9 shows a cross section of a magnetic storage medium.

FIG. 10 shows a cross section of a optically-readable data storage medium.

This invention provides certain crystallized, protein kinase-ligand complexes, in particular P38-ligand complexes, and their structure coordinates. The structure coordinates are based on the structure of a phosphorylated P38γ complex that has now been solved and which reveals new structural information regarding the activated states of other, related kinase proteins as described herein. The key structural features of the protein, particularly the shape of the substrate binding site, are useful in methods for designing inhibitors of the P38 and in solving the structures of other proteins with similar features.

As used herein, unless otherwise indicated, the term “p38γ” refers to the protein that is described by SEQ ID NO:1. In describing protein structure and function, reference is made to amino acids comprising the protein. The amino acids may also be referred to by their conventional abbreviations, as shown in the table below.

This invention also provides a crystallizable composition from which the crystallized protein is obtained. The crystallizable composition preferably comprises a phosphorylated P38 protein complexed with a substrate or ligand. The ligand may be any ligand capable of binding to the P38 protein, and is preferably a ligand that binds to the ATP binding site of the protein. Examples of such ligands are small molecule inhibitors of the particular P38 as well as non-hydrolyzable ATP analogs and suicide substrates. Non-hydrolyzable ATP analogs useful in the crystallizable compositions of this invention include AMP-PCH 2 P, AMP-PSP and AMP-PNP where the oxygen linking the second and third phosphates of the ATP analogs is replaced by CH 2 , S and NH, respectively. An example of a suicidal substrate is 5′-(p-fluorosulfonyl benzoyl)adenosine (FSBA). Preferably, the crystallizable compositions of this invention comprise AMP-PNP as the substrate. It is preferred that the composition further comprise divalent cations, especially magnesium or magnanese cations, which may be introduced in any suitable manner. For example, the cations may be introduced by incubating the desired ligand with a suitable metal salt such as MgCl 2 prior to incubation with the phosphorylated P38 protein.

It has been found that the crystallization of the phosphorylated P38 protein is sensitive to buffer conditions. Thus, in a preferred embodiment, the crystallizable compositions of this invention further comprise a suitable glycol such as ethylene glycol, polyethylene glycol (PEG), PEG-monomethyl ether or mixtures thereof, preferably PEG 4000, as an aqueous solution containing between about 10 to 35% of the glycol by volume of solution, a salt, such as sodium acetate at about 50 to 200 mM, a reducing agent, such as dithiothreitol (DTT) at between about 1 to 10 mM, a detergent such as C12E9 at about 0.01 to 0.05%, and a buffer that maintains pH at between about 8.0 and 9.0. An example of a suitable buffer is 100 mM Tris at pH 8.5.

By applying standard crystallization protocols to the above described crystallizable compositions, crystals of the phosphorylated P38 protein complex may be obtained. Thus, one aspect of this invention relates to a method of preparing phosphorylated P38-containing crystals. The method comprises the steps of

(a) obtaining a crystallizable composition comprising a phosphorylated P38 protein, divalent cations, and a ligand capable of binding to the protein, and

(b) subjecting the composition of step (a) to conditions which promote crystallization.

FIGS. 1 . 1 - 1 . 98 show the structure coordinates of a phosphorylated P38γ protein complexed with MgAMP-PNP. The manner of obtaining these structure coordinates, interpretation of the coordinates and their utility in understanding the protein structure, as described herein, will be understood by those of skill in the art and by reference to standard texts such as Crystal Structure Analysis, Jenny Pickworth Glusker and Kenneth N. Trueblood, 2nd Ed. Oxford University Press, 1985, New York; and Principles of Protein Structure, G. E. Schulz and R. H. Schirmer, Springer-Verlag, 1985, New York.

Those of skill in the art understand that a set of structure coordinates for an enzyme or an enzyme-complex or a portion thereof, is a relative set of points that define a shape in three dimensions. Thus, it is possible that an entirely different set of coordinates could define a similar or identical shape. Moreover, slight variations in the individual coordinates will have little effect on overall shape. In terms of binding pockets, these variations would not be expected to significantly alter the nature of ligands that could associate with those pockets.

These variations in coordinates may be generated because of mathematical manipulations of the P38γ/MgAMP-PNP structure coordinates. For example, the structure coordinates set forth in FIGS. 1 . 1 - 1 . 98 could be manipulated by crystallographic permutations of the structure coordinates, fractionalization of the structure coordinates, integer additions or subtractions to sets of the structure coordinates, inversion of the structure coordinates or any combination of the above.

Alternatively, modifications in the crystal structure due to mutations, additions, substitutions, and/or deletions of amino acids, or other changes in any of the components that make up the crystal could also account for variations in structure coordinates. If such variations are within an acceptable standard error as compared to the original coordinates, the resulting three-dimensional shape is considered to be the same. Thus, for example, a ligand that bound to the active site binding pocket of P38γ would also be expected to bind to another binding pocket whose structure coordinates defined a shape that fell within the acceptable error.

The term “binding pocket” refers to a region of the protein that, as a result of its shape, favorably associates with a ligand or substrate. The term “P38γ-like binding pocket” refers to a portion of a molecule or molecular complex whose shape is sufficiently similar to the P38γ binding pockets as to bind common ligands. This commonality of shape may be quantitatively defined by a root mean square deviation (rmsd) from the structure coordinates of the backbone atoms of the amino acids that make up the binding pockets in P38γ (as set forth in FIGS. 1 . 1 - 1 . 98 ). The method of performing this rmsd calculation is described below.

The “active site binding pockets” or “active site” of P38γ refers to the area on the P38γ enzyme surface where the substrate binds. In resolving the crystal structure of phosphorylated P38γ in complex with MgAMP-PNP, applicants have determined that P38γ amino acids Val33, Ala40, Val41, Ala54, Lys56, Ile87, Met109, Pro110, Phe111, Met112, Gly113, Thr114, Asp115, Lys118, Asp153, Lys155, Gly157, Asn158, Ala160, Leu170, Asp171, Gly173, and Leu174 are within 5 Å of and therefore close enough to interact with MgAMP-PNP. These amino acids are hereinafter referred to as the “SET 5A amino acids.” Thus, a binding pocket defined by the structural coordinates of those amino acids, as set forth in FIGS. 1 . 1 - 1 . 98 ; or a binding pocket whose root mean square deviation from the structure coordinates of the backbone atoms of those amino acids of not more than about 1.15 angstroms (Å) is considered a P38γ-like binding pocket of this invention.

Applicants have also determined that in addition to the P38γ amino acids set forth above, Pro32, Cys42, Ser43, Val53, Ile55, Lys57, Leu58, Thr59, Arg70, Glu74, Gly88, Leu107, Val108, Leu116, Gly117, Pro156, Leu159, Val161, Lys168, Phe172, Ala175, and Thr188 are within 8 Å of bound MgAMP-PNP and therefore are also close enough to interact with that substrate. These amino acids, in addition to the SET 5A amino acids, are hereinafter referred to as the “SET 8A amino acids.” Thus, in a preferred embodiment, a binding pocket defined by the structural coordinates of the amino acids within 8A of bound MgAMP-PNP, as set forth in FIGS. 1 . 1 - 1 . 98 ; or a binding pocket whose root mean square deviation from the structure coordinates of the backbone atoms of those amino acids of not more than about 1.15 Å is considered a preferred P38γ-like binding pocket of this invention.

It will be readily apparent to those of skill in the art that the numbering of amino acids in other isoforms of P38 may be different than that set forth for P38γ. Corresponding amino acids in other isoforms of P38 are easily identified by visual inspection of the amino acid sequences or by using commercially available homology software programs, as further described below.

Various computational analyses may be used to determine whether a protein or the binding pocket portion thereof is sufficiently similar to the P38γ binding pockets described above. Such analyses may be carried out in well known software applications, such as the Molecular Similarity application of QUANTA (Molecular Simulations Inc., San Diego, Calif.) version 4.1, and as described in the accompanying User's Guide.

For the purpose of this invention, a rigid fitting method was conveniently used to compare protein structures. Any molecule or molecular complex or binding pocket thereof having a root mean square deviation of conserved residue backbone atoms (N, Cα, C, O) of less than about 1.15 Å when superimposed on the relevant backbone atoms described by structure coordinates listed in FIGS. 1 . 1 - 1 . 98 are considered identical. More preferably, the root mean square deviation is less than about 1.0 Å.

The P38 X-ray coordinate data, when used in conjunction with a computer programmed with software to translate those coordinates into the 3-dimensional structure of p38γ may be used for a variety of purposes, especially for purposes relating to drug discovery. Such software for generating three-dimensional graphical representations are known and commercially available. The ready use of the coordinate data requires that it be stored in a computer-readable format. Thus, in accordance with the present invention, data capable of being displayed as the three dimensional structure of P38γ and portions thereof and their structurally similar homologues is stored in a machine-readable storage medium, which is capable of displaying a graphical three-dimensional representation of the structure.

Therefore, another embodiment of this invention provides a machine-readable data storage medium, comprising a data storage material encoded with machine readable data which, when used by a machine programmed with instructions for using said data, displays a graphical three-dimensional representation of a molecule or molecular complex comprising a binding pocket defined by structure coordinates of the P38γ SET 5A amino acids, or preferably the P38γ SET 8A amino acids, or a homologue of said molecule or molecular complex, wherein said homologue comprises a binding pocket that has a root mean square deviation from the backbone atoms of said amino acids of not more than about 1.15 Å.

Even more preferred is a machine-readable data storage medium that is capable of displaying a graphical three-dimensional representation of a molecule or molecular complex that is defined by the structure coordinates of all of the amino acids in FIG. 1 a or a homologue of said molecule or molecular complex, wherein said homologue has a root mean square deviation from the backbone atoms of all of the amino acids in FIGS. 1 . 1 - 1 . 98 of not more than about 1.15 Å.

According to an alternate embodiment, the machine-readable data storage medium comprises a data storage material encoded with a first set of machine readable data which comprises the Fourier transform of the structure coordinates set forth in FIGS. 1 . 1 - 1 . 98 , and which, when using a machine programmed with instructions for using said data, can be combined with a second set of machine readable data comprising the X-ray diffraction pattern of another molecule or molecular complex to determine at least a portion of the structure coordinates corresponding to the second set of machine readable data.

For example, the Fourier transform of the structure coordinates set forth in FIG. 1 a may be used to determine at least a portion of the structure coordinates of other P38s, such as P38β, and P38δ and isoforms of P38β, P38δ or P38γ. The structure coordinates in FIGS. 1 . 1 - 1 . 98 and the Fourier transform of the coordinates are especially useful for determining the coordinates of other P38s in phosphorylated form.

According to an alternate embodiment, this invention provides a computer for producing a three-dimensional representation of a molecule or molecular complex, wherein said molecule or molecular complex comprises a binding pocket defined by the P38γ SET 5A amino acids, or preferably the P38γ SET 8A amino acids, or a homologue of said molecule or molecular complex, wherein said homologue comprises a binding pocket that has a root mean square deviation from the backbone atoms of said amino acids of not more than 1.15 Å, wherein said computer comprises:

(a) a machine readable data storage medium comprising a data storage material encoded with machine-readable data, wherein said machine readable data comprises the structure coordinates of P38γ or portions thereof;

(b) a working memory for storing instructions for processing said machine-readable data;

(c) a central-processing unit coupled to said working memory and to said machine-readable data storage medium, for processing said machine-readable data into said three-dimensional representation; and

(d) an output hardware coupled to said central processing unit, for receiving said three Dimensional representation.

FIG. 8 demonstrates one version of these embodiments. System 10 includes a computer 11 comprising a central processing unit (“CPU”) 20 , a working memory 22 which may be, e.g., RAM (random-access memory) or “core” memory, mass storage memory 24 (such as one or more disk drives or CD-ROM drives), one or more cathode-ray tube (“CRT”) display terminals 26 , one or more keyboards 28 , one or more input lines 30 , and one or more output lines 40 , all of which are interconnected by a conventional bi-directional system bus 50 .

Input hardware 36 , coupled to computer 11 by input lines 30 , may be implemented in a variety of ways. Machine-readable data of this invention may be inputted via the use of a modem or modems 32 connected by a telephone line or dedicated data line 34 . Alternatively or additionally, the input hardware 36 may comprise CD-ROM drives or disk drives 24 . In conjunction with display terminal 26 , keyboard 28 may also be used as an input device.

Output hardware 46 , coupled to computer 11 by output lines 40 , may similarly be implemented by conventional devices. By way of example, output hardware 46 may include CRT display terminal 26 for displaying a graphical representation of a binding pocket of this invention using a program such as QUANTA as described herein. Output hardware might also include a printer 42 , so that hard copy output may be produced, or a disk drive 24 , to store system output for later use.

In operation, CPU 20 coordinates the use of the various input and output devices 36 , 46 coordinates data accesses from mass storage 24 and accesses to and from working memory 22 , and determines the sequence of data processing steps. A number of programs may be used to process the machine-readable data of this invention. Such programs are discussed in reference to the computational methods of drug discovery as described herein. Specific references to components of the hardware system 10 are included as appropriate throughout the following description of the data storage medium.

FIG. 9 shows a cross section of a magnetic data storage medium 100 which can be encoded with a machine-readable data that can be carried out by a system such as system 10 of FIG. 8 . Medium 100 can be a conventional floppy diskette or hard disk, having a suitable substrate 101 , which may be conventional, and a suitable coating 102 , which may be conventional, on one or both sides, containing magnetic domains (not visible) whose polarity or orientation can be altered magnetically. Medium 100 may also have an opening (not shown) for receiving the spindle of a disk drive or other data storage device 24 . The magnetic domains of coating 102 of medium 100 are polarized or oriented so as to encode in manner which may be conventional, machine readable data such as that described herein, for execution by a system such as system 10 of FIG. 8 .

FIG. 10 shows a cross section of an optically-readable data storage medium 110 which also can be encoded with such a machine-readable data, or set of instructions, which can be carried out by a system such as system 10 of FIG. 8 . Medium 110 can be a conventional compact disk read only memory (CD-ROM) or a rewritable medium such as a magneto-optical disk which is optically readable and magneto-optically writable. Medium 100 preferably has a suitable substrate 111 , which may be conventional, and a suitable coating 112 , which may be conventional, usually of one side of substrate 111 .

In the case of CD-ROM, as is well known, coating 112 is reflective and is impressed with a plurality of pits 113 to encode the machine-readable data. The arrangement of pits is read by reflecting laser light off the surface of coating 112 . A protective coating 114 , which preferably is substantially transparent, is provided on top of coating 112 .

In the case of a magneto-optical disk, as is well known, coating 112 has no pits 113 , but has a plurality of magnetic domains whose polarity or orientation can be changed magnetically when heated above a certain temperature, as by a laser (not shown). The orientation of the domains can be read by measuring the polarization of laser light reflected from coating 112 . The arrangement of the domains encodes the data as described above.

As mentioned above, the P38γ X-ray coordinate data is useful for screening and identifying drugs that inhibit P38, especially phosphorylated P38. For example, the structure encoded by the data may be computationally evaluated for its ability to associate with putative substrates or ligands. Such compounds that associate with p38γ may inhibit p38γ, and are potential drug candidates. Additionally or alternatively, the structure encoded by the data may be displayed in a graphical three-dimensional representation on a computer screen. This allows visual inspection of the structure, as well as visual inspection of the structure's association with the compounds.

Thus, according to another embodiment, this invention relates to a method for evaluating the potential of a compound to associate with a molecule or molecular complex comprising a binding pocket defined by the structure coordinates of the P38γ SET 5A amino acids, or preferably the P38γ SET 8A amino acids, or a homologue of said molecule or molecular complex, wherein said homologue comprises a binding pocket that has a root mean square deviation from the backbone atoms of said amino acids of not more than about 1.15 Å.

This method comprises the steps of:

a) creating a computer model of the binding pocket using structure coordinates wherein the root mean square deviation between said structure coordinates and the structure coordinates of the P38γ amino acids Val33, Ala40, Val41, Ala54, Lys56, Ile87, Met109, Pro110, Phe111, Met112, Gly113, Thr114, Asp115, Lys118, Asp153, Lys155, Gly157, Asn158, Ala160, Leu170, Asp171, Gly173, and Leu174 according to FIGS. 1 . 1 - 1 . 98 is not more than about 1.15 Å;

b) employing computational means to perform a fitting operation between the chemical entity and said computer model of the binding pocket; and

c) analyzing the results of said fitting operation to quantify the association between the chemical entity and the binding pocket model.

The term “chemical entity”, as used herein, refers to chemical compounds or ligands, complexes of at least two chemical compounds, and fragments of such compounds or complexes.

Even more preferably, the method evaluates the potential of a chemical entity to associate with a molecule or molecular complex defined by the structure coordinates of all of the P38γ amino acids, as set forth in FIGS. 1 . 1 - 1 . 98 , or a homologue of said molecule or molecular complex having a root mean square deviation from the backbone atoms of said amino acids of not more than 1.15 Å.

Alternatively, the structural coordinates of the P38γ binding pocket can be utilized in a method for identifying a potential agonist or antagonist of a molecule comprising a P38γ-like binding pocket. This method comprises the steps of:

(a) using atomic coordinates of the P38γ SET 5A amino acids ± a root mean square deviation from the backbone atoms of said amino acids of not more than about 1.15Å, to generate a three-dimensional structure of molecule comprising a P38γ-like binding pocket;

(b) employing said three-dimensional structure to design or select said potential agonist or antagonist;

(c) synthesizing said agonist or antagonist; and

(d) contacting said agonist or antagonist with said molecule to determine the ability of said potential agonist or antagonist to interact with said molecule.

More preferred is the use of the atomic coordinates of the P38γ SET 8A amino acids, ± a root mean square deviation from the backbone atoms of said amino acids of not more than 1.15 Å, to generate a three-dimensional structure of molecule comprising a p38γ-like binding pocket. Most preferred is when the atomic coordinates of all the amino acids of P38γ according to FIGS. 1 . 1 - 1 . 98 ± a root mean square deviation from the backbone atoms of said amino acids of not more than 1.15 Å, are used to generate a three-dimensional structure of molecule comprising a P38γ-like binding pocket.

For the first time, the present invention permits the use of molecular design techniques to identify, select or design potential inhibitors of p38, based on the structure of a phosphorylated p38γ-like binding pocket. Such a predictive model is valuable in light of the high costs associated with the preparation and testing of the many diverse compounds that may possibly bind to the p38 protein.

According to this invention, a potential p38 inhibitor may now be evaluated for its ability to bind a P38γ-like binding pocket prior to its actual synthesis and testing. If a proposed compound is predicted to have insufficient interaction or association with the binding pocket, preparation and testing of the compound is obviated. However, if the computer modeling indicates a strong interaction, the compound may then be obtained and tested for its ability to bind. Testing to confirm binding may be performed using assays such as described in Example 6.

A potential inhibitor of a P38γ-like binding pocket may be computationally evaluated by means of a series of steps in which chemical entities or fragments are screened and selected for their ability to associate with the P38γ-like binding pockets.

One skilled in the art may use one of several methods to screen chemical entities or fragments for their ability to associate with a P38γ-like binding pocket. This process may begin by visual inspection of, for example, a P38γ-like binding pocket on the computer screen based on the P38γ structure coordinates in FIGS. 1 . 1 - 1 . 98 or other coordinates which define a similar shape generated from the machine-readable storage medium. Selected fragments or chemical entities may then be positioned in a variety of orientations, or docked, within that binding pocket as defined above. Docking may be accomplished using software such as Quanta and Sybyl, followed by energy minimization 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 fragments or chemical entities. These include:

1. GRID (P. J. Goodford, “A Computational Procedure for Determining Energetically Favorable Binding Sites on Biologically Important Macromolecules”, J. Med. Chem., 28, pp. 849-857 (1985)). GRID is available from Oxford University, Oxford, UK.

2. MCSS (A. Miranker et al., “Functionality Maps of Binding Sites: A Multiple Copy Simultaneous Search Method.” Proteins: Structure, Function and Genetics, 11, pp. 29-34 (1991)). MCSS is available from Molecular Simulations, San Diego, Calif.

3. AUTODOCK (D. S. Goodsell et al., “Automated Docking of Substrates to Proteins by Simulated Annealing”, Proteins: Structure, Function, and Genetics, 8, pp. 195-202 (1990)). AUTODOCK is available from Scripps Research Institute, La Jolla, Calif.

4. DOCK (I. D. Kuntz et al., “A Geometric Approach to Macromolecule-Ligand Interactions”, J. Mol. Biol., 161, pp. 269-288 (1982)). DOCK is available from University of California, San Francisco, Calif.

Once suitable chemical entities or fragments have been selected, they can be designed or 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 P38γ. This would be followed by manual model building using software such as Quanta or Sybyl [Tripos Associates, St. Louis, Mo].

Useful programs to aid one of skill in the art in connecting the individual chemical entities or fragments include:

1. CAVEAT (P. A. Bartlett et al, “CAVEAT: A Program to Facilitate the Structure-Derived Design of Biologically Active Molecules”, in Molecular Recognition in Chemical and Biological Problems”, Special Pub., Royal Chem. Soc., 78, pp. 182-196 (1989); G. Lauri and P. A. Bartlett, “CAVEAT: a Program to Facilitate the Design of Organic Molecules”, J. Comput. Aided Mol. Des., 8, pp. 51-66 (1994)). CAVEAT is available from the University of California, Berkeley, Calif.

2. 3D Database systems such as ISIS (MDL Information Systems, San Leandro, Calif.). This area is reviewed in Y. C. Martin, “3D Database Searching in Drug Design”, J. Med. Chem., 35, pp. 2145-2154 (1992).

3. HOOK (M. B. Eisen et al, “HOOK: A Program for Finding Novel Molecular Architectures that Satisfy the Chemical and Steric Requirements of a Macromolecule Binding Site”, Proteins: Struct., Funct., Genet., 19, pp. 199-221 (1994). HOOK is available from Molecular Simulations, San Diego, Calif.

Instead of proceeding to build an inhibitor of a P38γ-like binding pocket in a step-wise fashion one fragment or chemical entity at a time as described above, inhibitory or other P38γ binding compounds may be designed as a whole or “de novo” using either an empty binding site or optionally including some portion(s) of a known inhibitor(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.).

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 modeling techniques may also be employed in accordance with this invention [see, e.g., 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)].

Once a compound has been designed or selected by the above methods, the efficiency with which that entity may bind to a P38γ binding pocket may be tested and optimized by computational evaluation. For example, an effective P38γ binding pocket inhibitor must preferably demonstrate a relatively small difference in energy between its bound and free states (i.e., a small deformation energy of binding). Thus, the most efficient P38γ binding pocket inhibitors should preferably be designed with a deformation energy of binding of not greater than about 10 kcal/mole, more preferably, not greater than 7 kcal/mole. P38γ binding pocket inhibitors may interact with the binding pocket in more than one of multiple conformations that are 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 entity and the average energy of the conformations observed when the inhibitor binds to the protein.

An entity designed or selected as binding to a P38γ binding pocket may be further computationally optimized so that in its bound state it would preferably lack repulsive electrostatic interaction with the target enzyme and with the surrounding water molecules. Such non-complementary electrostatic interactions include repulsive charge-charge, dipole-dipole and charge-dipole interactions.

Specific computer software is available in the art to evaluate compound deformation energy and electrostatic interactions. Examples of programs designed for such uses include: Gaussian 94, revision C (M. J. Frisch, Gaussian, Inc., Pittsburgh, Pa. ©1995); AMBER, version 4.1 (P. A. Kollman, University of California at San Francisco, ©1995); QUANTA/CHARMM (Molecular Simulations, Inc., San Diego, Calif. ©1995); Insight II/Discover (Molecular Simulations, Inc., San Diego, Calif. ©1995); DelPhi (Molecular Simulations, Inc., San Diego, Calif. ©1995); and AMSOL (Quantum Chemistry Program Exchange, Indiana University). These programs may be implemented, for instance, using a Silicon Graphics workstation such as an Indigo 2 with “IMPACT” graphics. Other hardware systems and software packages will be known to those skilled in the art.

Another approach enabled by this invention, is the computational screening of small molecule databases for chemical entities or compounds that can bind in whole, or in part, to a P38γ binding pocket. In this screening, the quality of fit of such entities to the binding site may be judged either by shape complementarity or by estimated interaction energy [E. C. Meng et al., J. Comp. Chem., 13, 505-524 (1992)].

According to another embodiment, the invention provides compounds which associate with a P38γ-like binding pocket produced or identified by the method set forth above.

The structure coordinates set forth in FIGS. 1 . 1 - 1 . 98 can also be used to aid in obtaining structural information about another crystallized molecule or molecular complex. This may be achieved by any of a number of well-known techniques, including molecular replacement.

Therefore, in another embodiment this invention provides a method of utilizing molecular replacement to obtain structural information about a molecule or molecular complex whose structure is unknown comprising the steps of:

a) crystallizing said molecule or molecular complex of unknown structure;

b) generating an X-ray diffraction pattern from said crystallized molecule or molecular complex; and

c) applying at least a portion of the structure coordinates set forth in FIGS. 1 . 1 - 1 . 98 to the X-ray diffraction pattern to generate a three-dimensional electron density map of the molecule or molecular complex whose structure is unknown.

By using molecular replacement, all or part of the structure coordinates of the P38γ/MgAMP-PNP complex as provided by this invention (and set forth in FIGS. 1 . 1 - 1 . 98 ) can be used to determine the structure of another crystallized molecule or molecular complex more quickly and efficiently than attempting an ab initio structure determination.

Molecular replacement provides an accurate estimation of the phases for an unknown structure. Phases are a factor in equations used to solve crystal structures that can not be determined directly. Obtaining accurate values for the phases, by methods other than molecular replacement, is a time-consuming process that involves iterative cycles of approximations and refinements and greatly hinders the solution of crystal structures. However, when the crystal structure of a protein containing at least a homologous portion has been solved, the phases from the known structure provide a satisfactory estimate of the phases for the unknown structure.

Thus, this method involves generating a preliminary model of a molecule or molecular complex whose structure coordinates are unknown, by orienting and positioning the relevant portion of the P38γ/MgAMP-PNP complex according to FIGS. 1 . 1 - 1 . 98 within the unit cell of the crystal of the unknown molecule or molecular complex so as best to account for the observed X-ray diffraction pattern of the crystal of the molecule or molecular complex whose structure is unknown. Phases can then be calculated from this model and combined with the observed X-ray diffraction pattern amplitudes to generate an electron density map of the structure whose coordinates are unknown. This, in turn, can be subjected to any well-known model building and structure refinement techniques to provide a final, accurate structure of the unknown crystallized molecule or molecular complex [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)].

The structure of any portion of any crystallized molecule or molecular complex that is sufficiently homologous to any portion of the P38γ/MgAMP-PNP complex can be resolved by this method.

In a preferred embodiment, the method of molecular replacement is utilized to obtain structural information about another P38, such as P38α, P38β, P38δ, or isoforms of P38β, P38δ or P38γ. The structure coordinates of P38γ as provided by this invention are particularly useful in solving the structure of other isoforms of P38γ or P38γ complexes.

Furthermore, the structure coordinates of P38γ as provided by this invention are useful in solving the structure of P38γ proteins that have amino acid substitutions, additions and/or deletions (referred to collectively as “P38γ mutants”, as compared to naturally occurring P38γ isoforms). These P38γ mutants may optionally be crystallized in co-complex with a chemical entity, such as a non-hydrolyzable ATP analogue or a suicide substrate. The crystal structures of a series of such complexes may then be solved by molecular replacement and compared with that of wild-type p38γ. Potential sites for modification within the various binding sites of the enzyme may thus be identified. This information provides an additional tool for determining the most efficient binding interactions such as, for example, increased hydrophobic interactions, between P38γ and a chemical entity or compound.

All of the complexes referred to above may be studied using well-known X-ray diffraction techniques and may be refined versus 1.5-3A resolution X-ray data to an R value of about 0.22 or less using computer software, such as X-PLOR [Yale University, ©1992, distributed by Molecular Simulations, Inc.; see, e.g., Blundell & Johnson, supra; Meth. Enzymol., vol. 114 & 115, H. W. Wyckoff et al., eds., Academic Press (1985)]. This information may thus be used to optimize known P38γ inhibitors, and more importantly, to design new P38γ inhibitors.

The structure coordinates described above may also be used to derive the dihedral angles, φ and ψ, that define the conformation of the amino acids in the protein backbone. As will be understood by those skilled in the art, the φ n angle refers to the rotation around the bond between the alpha carbon and the nitrogen, and the ψ n angle refers to the rotation around the bond between the carbonyl carbon and the alpha carbon. The subscript “n” identifies the amino acid whose conformation is being described [for a general reference, see Blundell and Johnson, Protein Crystallography, Academic Press, London, 1976].

Surprisingly, it has now been found that for the crystalline P38γ-ligand complex, the conformation of Gly113 is very different from the conformations reported for corresponding amino acids in other protein kinases. In order to compare the conformations of P38γ and other protein kinases at a particular amino acid site, such as Gly113, along the polypeptide backbone well-known procedures may be used for doing sequence alignments of the amino acids. Such sequence alignments allow for the equivalent or corresponding sites to be compared. One such method for doing a sequence alignment is the “bestfit” program available from Genetics Computer Group which uses the local homology algorithm described by Smith and Waterman in Advances in Applied Mathematics 2; 482 (1981).

A suitable amino acid sequence alignment will require that the proteins being aligned share a minimum percentage of identical amino acids. Generally, a first protein being aligned with a second protein should share in excess of about 35% identical amino acids with the second protein. Hanks et al., Science, 241, 42 (1988); Hanks and Quinn, Methods in Enzymology, 200, 38 (1991).

Equivalents of the Gly113 residue of p38γ may also be identified by its functional position. Gly113 is the amino acid residue that immediately follows sequentially the amino acid residue that donates, or is capable of donating, a hydrogen bond to the N1 nitrogen of the adenosine ring of ATP or an ATP analog, if such ATP or ATP analog were to be in the binding pocket comprising the Gly113 residue. The ability of the amino acid to donate such a hydrogen bond occurs as the result of the spatial position of the amino acid in the binding pocket of the protein. As used herein, the term “corresponding amino acid” or “equivalent amino acid” refers to a particular amino acid in a protein kinase that corresponds to another, particular amino acid in a different protein kinase as determined by sequence alignment and/or its functional position.

Table 1 shows the sequence alignments for selected protein kinases where corresponding amino acids are shown in the same column. The amino acid numbering is based on the assignments given in the Swiss-Prot database which is an international protein sequence database distributed by the European Bioinformatics Institute (EBI) in Geneva, Switzerland. The database can be found at www.ebi.ac.uk/swissprot. Erk6_HUMAN is the database protein name for P38γ. The ten amino acids immediately preceding G113 of P38γ are given starting with T103. Thus, for example, Gly113 of P38γ corresponds or is equivalent to the following: Gly110 of P38α (MP38_HUMAN), Glu107 of mouse ERK2, and Asp150 of human JNK3. The last column of Table 1 shows the Swiss-Prot database accession number.

As noted above, the conformation of Gly113 is very different from the conformations reported for corresponding amino acids in other protein kinases. For Gly113 of the P38γ-AMPPNP complex, ψ 112 was found to be about 24 degrees and φ 113 was found to be about 96 degrees. Table 2 shows the dihedral angles for Met112 and Gly113 of P38γ-AMPPNP complex and how these angles compare to those of the corresponding amino acids in other MAP kinases whose crystal structures have been reported. The protein names for the known proteins are provided as their Protein Data Banks™ (pdb) accession numbers. The Protein Data Bank is an international repository for three dimensional structures and can be located at www.rcsb.org/pdb/.

It is well-recognized that there will be some variability in the conformations of corresponding amino acids in similar or identical proteins when the protein crystallization and structure determination are repeated. This variability in the φ and ψ dihedral angles may be approximated by reference to Ramachandran plots comparing the conformations obtained for two or more identical or similar proteins [Blundell and Johnson, Protein Crystallography, Academic Press, London, 1976]. It may be expected that the dihedral angles of equivalent amino acid residues in identical or similar proteins will vary as much as about 45° or more.

It should be noted that the amino acid numbering defined in the Protein Data Banks™ may be offset from the numbering given in the Swiss-Prot database. This offset, when it occurs, will be readily understood by those skilled in the art. Thus, the sequences of those proteins that are listed in both databases may be easily compared despite offsets in amino acid numbering that may occur. Examples of such offsets occur for INSR_HUMAN where A1107 according to Swiss-Prot numbering is the same as A1080 in the PDB database and for LCK_HUMAN where E319 according to Swiss-Prot numbering is the same as E320 by PDB numbering.

The ψ 112 and φ 113 dihedral angles of the P38γ-AMPPNP complex shown in Table 2 indicate that the conformation of Gly113 in this complex is “flipped” or rotated considerably relative to corresponding amino acids in other MAP kinases. Therefore, the structure coordinates of P38γ set forth in FIGS. 1 . 1 - 1 . 98 represent, inter alia, what is believed to be a conformation at Met 112 and Gly113 that had not been observed for other crystalline protein kinases, especially other MAP kinases.

Accordingly, another embodiment of this invention relates to a crystalline protein kinase-ligand complex, said kinase comprising amino acid residues that correspond by functional and/or sequence alignment to the Met112 and Gly113 residues of P38γ or that correspond by functional and/or sequence alignment to the Met112 and Gly113 equivalent residues of one or more proteins listed in Table 1, wherein the ψ angle of the residue corresponding to Met112 is in the range of about −60° to 60° and the ψ angle of the residue corresponding to Gly113 is in the range of about 30° to 150°. Preferably, the ψ angle of the crystalline protein kinase-ligand complex is in the range of about −45° to 45° and most preferably in the range of about −30° to 30°. Preferably, the φ angle is in the range of about 45° to 135°, and most preferably is in the range of about 60° to 120°. Examples of kinases that may provide such a crystalline protein kinase when complexed with a ligand are described by Hanks et al., Science, 241, 42 (1988) and Hanks and Quinn, Methods in Enzymology, 200, 38 (1991). Other examples of such kinases may be found at www.sdsc.edu/Kinases/pkr/pk_catalytic/pk_hanks_seq_align_long.html, where the kinases are listed with their corresponding sequence alignments.

Another embodiment of this invention relates to a crystalline protein kinase-ligand complex, said kinase selected from the proteins listed in Table 1, wherein the ψ angle of the residue corresponding to Met112 is in the range of about −60° to 60° and the φ angle of the residue corresponding to Gly113 is in the range of about 30° to 150°. Preferably, the ψ angle of the crystalline protein kinase-ligand complex is in the range of about −45° to 45° and most preferably in the range of about −30° to 30°. Preferably, the φ angle is in the range of about 45° to 135°, and most preferably is in the range of about 60° to 120°.

Structural information regarding the conformation of the Met112 and Gly113 residues of the crystalline P38γ complex may be encoded in a machine-readable data storage medium as described above for encoding the other structural coordinates of the protein. Accordingly, another embodiment of this invention relates to a computer for producing a three-dimensional representation of an ATP binding site of a protein kinase-ligand complex, or portion thereof, wherein said computer comprises:

a) a machine-readable data storage medium comprising a data storage material encoded with machine-readable data, wherein said machine-readable data comprises the structure coordinates of a kinase, or portion thereof, said kinase or portion thereof comprising amino acid residues that correspond by functional and/or sequence alignment to the Met112 and Gly113 residues of P38γ or that correspond by functional and/or sequence alignment to the Met112 and Gly113 equivalent residues of one or more proteins listed in Table 1, wherein the ψ angle of the residue corresponding to Met112 is in the range of about −60° to 60° and the φ angle of the residue corresponding to Gly113 is in the range of about 30° to 150°;

b) a working memory for storing instructions for processing said machine-readable data;

c) a central-processing unit coupled to said working memory and to said machine-readable data storage medium, for processing said machine readable data into said three-dimensional representation; and

d) an output hardware coupled to said central-processing unit, for receiving said three-dimensional representation. Preferably, the machine-readable data comprises the structure coordinates of a kinase, or portion thereof, said kinase comprising amino acid residues corresponding to the Met112 and Gly113 amino acids of P38γ or corresponding to the Met112 and Gly113 equivalent residues of one or more proteins listed in Table 1, wherein the ψ angle is in the range of about −45° to 45° and most preferably in the range of about −30° to 30°, and the φ angle is in the range of about 45° to 135° and most preferably in the range of about 60° to 120°. In a more preferred embodiment of this computer, the machine readable data comprises the structure coordinates of a crystalline protein kinase-ligand complex, or portion thereof, where said kinase is selected from a protein listed in Table 1.

For designing new compounds that associate with a protein kinase binding pocket, it is useful to employ information that includes the conformations of the Met112 and Gly113 residues, or their equivalents, along with other structural information regarding amino acids in the binding pocket. For example, to evaluate the ability of a chemical entity to bind to a protein kinase, the conformations of Met112 and Gly113, or equivalents, may be used along with structure coordinates of the backbone atoms of amino acids in the protein kinase binding pocket. These structure coordinates and the structure coordinates of the p38γ amino acids Val33, Ala40, Val41, Ala54, Lys56, Ile87, Met109, Pro110, Phe111, Met112, Gly113, Thr114, Asp115, Lys118, Asp153, Lys155, Gly157, Asn158, Ala160, Leu170, Asp171, Gly173, and Leu174 according to FIGS. 1 . 1 - 1 . 98 should not differ by more than about 3.0 angstroms in root mean square deviation, preferably the root mean square deviation is within about 2.7 angstroms, and most preferably within about 2.5 angstroms. For example, the root mean square deviation between the structure coordinates of the p38γ amino acids and those of a p38γ complex (see Table 2) was found by applicants to be 2.41 angstroms. Resolution error may account for variation in the root mean square deviation of a few tenths of an angstrom.

Accordingly, another embodiment of this invention provides a method for evaluating the ability of a chemical entity to associate with a protein kinase binding pocket, said method comprising the steps of:

a) creating a computer model of the binding pocket using structure coordinates wherein:

(i) the root mean square deviation between said structure coordinates and the structure coordinates of the P38γ amino acids Val33, Ala40, Val41, Ala54, Lys56, Ile87, Met109, Pro110, Phe111, Met112, Gly113, Thr114, Asp115, Lys118, Asp153, Lys155, Gly157, Asn158, Ala160, Leu170, Asp171, Gly173, and Leu174 according to FIGS. 1 . 1 - 1 . 98 is within about 3.0 angstroms,

(ii) said binding pocket model depicts amino acid residues that correspond by functional and/or sequence alignment to the Met112 and Gly113 residues of P38γ or that correspond by functional and/or sequence alignment to the Met112 and Gly113 equivalent residues of one or more proteins listed in Table 1, and

(iii) said binding pocket model depicts the ψ angle of the residue corresponding to Met112 to be in the range of about −60° to 60° and the φ angle of the residue corresponding to Gly113 to be in the range of about 30° to 150°;

b) employing computational means to perform a fitting operation between the chemical entity and the binding pocket model; and

c) analyzing the results of said fitting operation to quantify the association between the chemical entity and the binding pocket model.

A useful root mean square deviation between the structure coordinates of a particular binding pocket and the structure coordinates of the binding pocket of another protein kinase may be readily determined by one skilled in the art. For example, when the protein kinase is selected from a protein listed in Table 1, the root mean square deviation is preferably within about 2.7 angstroms, and is more preferably within about 2.5 angstroms.

This invention also provides a method for identifying a potential agonist or antagonist of a molecule comprising a P38γ-like binding pocket, comprising the steps of:

a) creating a computer model of the binding pocket using structure coordinates wherein:

(i) the root mean square deviation between said structure coordinates and the structure coordinates of the P38γ amino acids Val33, Ala40, Val41, Ala54, Lys56, Ile87, Met109, Pro110, Phe111, Met112, Gly113, Thr114, Asp115, Lys118, Asp153, Lys155, Gly157, Asn158, Ala160, Leu170, Asp170, Gly173, and Leu174 according to FIGS. 1 . 1 - 1 . 98 is within about 3.0 angstroms,

(ii) said binding pocket model depicts amino acid residues that correspond by functional and/or sequence alignment to the Met112 and Gly113 residues of P38γ or that correspond by functional and/or sequence alignment to the Met112 and Gly113 equivalent residues of one or more proteins listed in Table 1, and

(iii) said binding pocket model depicts the ψ angle of the residue corresponding to Met112 to be in the range of about −60° to 60° and the φ angle of the residue corresponding to Gly113 to be in the range of about 30° to 150°;

b) employing said model of the binding pocket to design or select said potential agonist or antagonist;

c) synthesizing said agonist or antagonist; and

d) contacting said agonist or antagonist with said molecule to determine the ability of said potential agonist or antagonist to interact with said molecule.

A preferred embodiment of this method uses the structure coordinates of the Met112 and Gly113 amino acids of p38γ or the Met112 and Gly113 equivalent residues of a protein listed in Table 1.

In order that this invention be more fully understood, the following examples are set forth. These examples are for the purpose of illustration only and are not to be construed as limiting the scope of the invention in any way.

Human

1

Met Ser Ser Pro Pro Pro Ala Arg Ser Gly Phe Tyr Arg Gln Glu Val

1 5 10 15

Thr Lys Thr Ala Trp Glu Val Arg Ala Val Tyr Arg Asp Leu Gln Pro

20 25 30

Val Gly Ser Gly Ala Tyr Gly Ala Val Cys Ser Ala Val Asp Gly Arg

35 40 45

Thr Gly Ala Lys Val Ala Ile Lys Lys Leu Tyr Arg Pro Phe Gln Ser

50 55 60

Glu Leu Phe Ala Lys Arg Ala Tyr Arg Glu Leu Arg Leu Leu Lys His

65 70 75 80

Met Arg His Glu Asn Val Ile Gly Leu Leu Asp Val Phe Thr Pro Asp

85 90 95

Glu Thr Leu Asp Asp Phe Thr Asp Phe Tyr Leu Val Met Pro Phe Met

100 105 110

Gly Thr Asp Leu Gly Lys Leu Met Lys His Glu Lys Leu Gly Glu Asp

115 120 125

Arg Ile Gln Phe Leu Val Tyr Gln Met Leu Lys Gly Leu Arg Tyr Ile

130 135 140

His Ala Ala Gly Ile Ile His Arg Asp Leu Lys Pro Gly Asn Leu Ala

145 150 155 160

Val Asn Glu Asp Cys Glu Leu Lys Ile Leu Asp Phe Gly Leu Ala Arg

165 170 175

Gln Ala Asp Ser Glu Met Thr Gly Tyr Val Val Thr Arg Trp Tyr Arg

180 185 190

Ala Pro Glu Val Ile Leu Asn Trp Met Arg Tyr Thr Gln Thr Val Asp

195 200 205

Ile Trp Ser Val Gly Cys Ile Met Ala Glu Met Ile Thr Gly Lys Thr

210 215 220

Leu Phe Lys Gly Ser Asp His Leu Asp Gln Leu Lys Glu Ile Met Lys

225 230 235 240

Val Thr Gly Thr Pro Pro Ala Glu Phe Val Gln Arg Leu Gln Ser Asp

245 250 255

Glu Ala Lys Asn Tyr Met Lys Gly Leu Pro Glu Leu Glu Lys Lys Asp

260 265 270

Phe Ala Ser Ile Leu Thr Asn Ala Ser Pro Leu Ala Val Asn Leu Leu

275 280 285

Glu Lys Met Leu Val Leu Asp Ala Glu Gln Arg Val Thr Ala Gly Glu

290 295 300

Ala Leu Ala His Pro Tyr Phe Glu Ser Leu His Asp Thr Glu Asp Glu

305 310 315 320

Pro Gln Val Gln Lys Tyr Asp Asp Ser Phe Asp Asp Val Asp Arg Thr

325 330 335

Leu Asp Glu Trp Lys Arg Val Thr Tyr Lys Glu Val Leu Ser Phe Lys

340 345 350

Pro Pro Arg Gln Leu Gly Ala Arg Val Ser Lys Glu Thr Pro Leu

355 360 365

2

360

Human

2

Met Ser Gln Glu Arg Pro Thr Phe Tyr Arg Gln Glu Leu Asn Lys Thr

1 5 10 15

Ile Trp Glu Val Pro Glu Arg Tyr Gln Asn Leu Ser Pro Val Gly Ser

20 25 30

Gly Ala Tyr Gly Ser Val Cys Ala Ala Phe Asp Thr Lys Thr Gly Leu

35 40 45

Arg Val Ala Val Lys Lys Leu Ser Arg Pro Phe Gln Ser Ile Ile His

50 55 60

Ala Lys Arg Thr Tyr Arg Glu Leu Arg Leu Leu Lys His Met Lys His

65 70 75 80

Glu Asn Val Ile Gly Leu Leu Asp Val Phe Thr Pro Ala Arg Ser Leu

85 90 95

Glu Glu Phe Asn Asp Val Tyr Leu Val Thr His Leu Met Gly Ala Asp

100 105 110

Leu Asn Asn Ile Val Lys Cys Gln Lys Leu Thr Asp Asp His Val Gln

115 120 125

Phe Leu Ile Tyr Gln Ile Leu Arg Gly Leu Lys Tyr Ile His Ser Ala

130 135 140

Asp Ile Ile His Arg Asp Leu Lys Pro Ser Asn Leu Ala Val Asn Glu

145 150 155 160

Asp Cys Glu Leu Lys Ile Leu Asp Phe Gly Leu Ala Arg His Thr Asp

165 170 175

Asp Glu Met Thr Gly Tyr Val Ala Thr Arg Trp Tyr Arg Ala Pro Glu

180 185 190

Ile Met Leu Asn Trp Met His Tyr Asn Gln Thr Val Asp Ile Trp Ser

195 200 205

Val Gly Cys Ile Met Ala Glu Leu Leu Thr Gly Arg Thr Leu Phe Pro

210 215 220

Gly Thr Asp His Ile Asp Gln Leu Lys Leu Ile Leu Arg Leu Val Gly

225 230 235 240

Thr Pro Gly Ala Glu Leu Leu Lys Lys Ile Ser Ser Glu Ser Ala Arg

245 250 255

Asn Tyr Ile Gln Ser Leu Thr Gln Met Pro Lys Met Asn Phe Ala Asn

260 265 270

Val Phe Ile Gly Ala Asn Pro Leu Ala Val Asp Leu Leu Glu Lys Met

275 280 285

Leu Val Leu Asp Ser Asp Lys Arg Ile Thr Ala Ala Gln Ala Leu Ala

290 295 300

His Ala Tyr Phe Ala Gln Tyr His Asp Pro Asp Asp Glu Pro Val Ala

305 310 315 320

Asp Pro Tyr Asp Gln Ser Phe Glu Ser Arg Asp Leu Leu Ile Asp Glu

325 330 335

Trp Lys Ser Leu Thr Tyr Asp Glu Val Ile Ser Phe Val Pro Pro Pro

340 345 350

Leu Asp Gln Glu Glu Met Glu Ser

355 360

3

360

Human

3

Met Ala Ala Ala Ala Ala Ala Gly Ala Gly Pro Glu Met Val Arg Gly

1 5 10 15

Gln Val Phe Asp Val Gly Pro Arg Tyr Thr Asn Leu Ser Tyr Ile Gly

20 25 30

Glu Gly Ala Tyr Gly Met Val Cys Ser Ala Tyr Asp Asn Val Asn Lys

35 40 45

Val Arg Val Ala Ile Lys Lys Ile Ser Pro Phe Glu His Gln Thr Tyr

50 55 60

Cys Gln Arg Thr Leu Arg Glu Ile Lys Ile Leu Leu Arg Phe Arg His

65 70 75 80

Glu Asn Ile Ile Gly Ile Asn Asp Ile Ile Arg Ala Pro Thr Ile Glu

85 90 95

Gln Met Lys Asp Val Tyr Ile Val Gln Asp Leu Met Glu Thr Asp Leu

100 105 110

Tyr Lys Leu Leu Lys Thr Gln His Leu Ser Asn Asp His Ile Cys Tyr

115 120 125

Phe Leu Tyr Gln Ile Leu Arg Gly Leu Lys Tyr Ile His Ser Ala Asn

130 135 140

Val Leu His Arg Asp Leu Lys Pro Ser Asn Leu Leu Leu Asn Thr Thr

145 150 155 160

Cys Asp Leu Lys Ile Cys Asp Phe Gly Leu Ala Arg Val Ala Asp Pro

165 170 175

Asp His Asp His Thr Gly Phe Leu Thr Glu Tyr Val Ala Thr Arg Trp

180 185 190

Tyr Arg Ala Pro Glu Ile Met Leu Asn Ser Lys Gly Tyr Thr Lys Ser

195 200 205

Ile Asp Ile Trp Ser Val Gly Cys Ile Leu Ala Glu Met Leu Ser Asn

210 215 220

Arg Pro Ile Phe Pro Gly Lys His Tyr Leu Asp Gln Leu Asn His Ile

225 230 235 240

Leu Gly Ile Leu Gly Ser Pro Ser Gln Glu Asp Leu Asn Cys Ile Ile

245 250 255

Asn Leu Lys Ala Arg Asn Tyr Leu Leu Ser Leu Pro His Lys Asn Lys

260 265 270

Val Pro Trp Asn Arg Leu Phe Pro Asn Ala Asp Ser Lys Ala Leu Asp

275 280 285

Leu Leu Asp Lys Met Leu Thr Phe Asn Pro His Lys Arg Ile Glu Val

290 295 300

Glu Gln Ala Leu Ala His Pro Tyr Leu Glu Gln Tyr Tyr Asp Pro Ser

305 310 315 320

Asp Glu Pro Ile Ala Glu Ala Pro Phe Lys Phe Asp Met Glu Leu Asp

325 330 335

Asp Leu Pro Lys Glu Lys Leu Lys Glu Leu Ile Phe Glu Glu Thr Ala

340 345 350

Arg Phe Gln Pro Gly Tyr Arg Ser

355 360

4

358

Mouse

4

Met Ala Ala Ala Ala Ala Ala Gly Pro Glu Met Val Arg Gly Gln Val

1 5 10 15

Phe Asp Val Gly Pro Arg Tyr Thr Asn Leu Ser Tyr Ile Gly Glu Gly

20 25 30

Ala Tyr Gly Met Val Cys Ser Ala Tyr Asp Asn Leu Asn Lys Val Arg

35 40 45

Val Ala Ile Lys Lys Ile Ser Pro Phe Glu His Gln Thr Tyr Cys Gln

50 55 60

Arg Thr Leu Arg Glu Ile Lys Ile Leu Leu Arg Phe Arg His Glu Asn

65 70 75 80

Ile Ile Gly Ile Asn Asp Ile Ile Arg Ala Pro Thr Ile Glu Gln Met

85 90 95

Lys Asp Val Tyr Ile Val Gln Asp Leu Met Glu Thr Asp Leu Tyr Lys

100 105 110

Leu Leu Lys Thr Gln His Leu Ser Asn Asp His Ile Cys Tyr Phe Leu

115 120 125

Tyr Gln Ile Leu Arg Gly Leu Lys Tyr Ile His Ser Ala Asn Val Leu

130 135 140

His Arg Asp Leu Lys Pro Ser Asn Leu Leu Leu Asn Thr Thr Cys Asp

145 150 155 160

Leu Lys Ile Cys Asp Phe Gly Leu Ala Arg Val Ala Asp Pro Asp His

165 170 175

Asp His Thr Gly Phe Leu Thr Glu Tyr Val Ala Thr Arg Trp Tyr Arg

180 185 190

Ala Pro Glu Ile Met Leu Asn Ser Lys Gly Tyr Thr Lys Ser Ile Asp

195 200 205

Ile Trp Ser Val Gly Cys Ile Leu Ala Glu Met Leu Ser Asn Arg Pro

210 215 220

Ile Phe Pro Gly Lys His Tyr Leu Asp Gln Leu Asn His Ile Leu Gly

225 230 235 240

Ile Leu Gly Ser Pro Ser Gln Glu Asp Leu Asn Cys Ile Ile Asn Leu

245 250 255

Lys Ala Arg Asn Tyr Leu Leu Ser Leu Pro His Lys Asn Lys Val Pro

260 265 270

Trp Asn Arg Leu Phe Pro Asn Ala Asp Ser Lys Ala Leu Asp Leu Leu

275 280 285

Asp Lys Met Leu Thr Phe Asn Pro His Lys Arg Ile Glu Val Glu Gln

290 295 300

Ala Leu Ala His Pro Tyr Leu Glu Gln Tyr Tyr Asp Pro Ser Asp Glu

305 310 315 320

Pro Ile Ala Glu Ala Pro Phe Lys Phe Asp Met Glu Leu Asp Asp Leu

325 330 335

Pro Lys Glu Lys Leu Lys Glu Leu Ile Phe Glu Glu Thr Ala Arg Phe

340 345 350

Gln Pro Gly Tyr Arg Ser

355

5

464

Human

5

Met Ser Leu His Phe Leu Tyr Tyr Cys Ser Glu Pro Thr Leu Asp Val

1 5 10 15

Lys Ile Ala Phe Cys Gln Gly Phe Asp Lys Gln Val Asp Val Ser Tyr

20 25 30

Ile Ala Lys His Tyr Asn Met Ser Lys Ser Lys Val Asp Asn Gln Phe

35 40 45

Tyr Ser Val Glu Val Gly Asp Ser Thr Phe Thr Val Leu Lys Arg Tyr

50 55 60

Gln Asn Leu Lys Pro Ile Gly Ser Gly Ala Gln Gly Ile Val Cys Ala

65 70 75 80

Ala Tyr Asp Ala Val Leu Asp Arg Asn Val Ala Ile Lys Lys Leu Ser

85 90 95

Arg Pro Phe Gln Asn Gln Thr His Ala Lys Arg Ala Tyr Arg Glu Leu

100 105 110

Val Leu Met Lys Cys Val Asn His Lys Asn Ile Ile Ser Leu Leu Asn

115 120 125

Val Phe Thr Pro Gln Lys Thr Leu Glu Glu Phe Gln Asp Val Tyr Leu

130 135 140

Val Met Glu Leu Met Asp Ala Asn Leu Cys Gln Val Ile Gln Met Glu

145 150 155 160

Leu Asp His Glu Arg Met Ser Tyr Leu Leu Tyr Gln Met Leu Cys Gly

165 170 175

Ile Lys His Leu His Ser Ala Gly Ile Ile His Arg Asp Leu Lys Pro

180 185 190

Ser Asn Ile Val Val Lys Ser Asp Cys Thr Leu Lys Ile Leu Asp Phe

195 200 205

Gly Leu Ala Arg Thr Ala Gly Thr Ser Phe Met Met Thr Pro Tyr Val

210 215 220

Val Thr Arg Tyr Tyr Arg Ala Pro Glu Val Ile Leu Gly Met Gly Tyr

225 230 235 240

Lys Glu Asn Val Asp Ile Trp Ser Val Gly Cys Ile Met Gly Glu Met

245 250 255

Val Arg His Lys Ile Leu Phe Pro Gly Arg Asp Tyr Ile Asp Gln Trp

260 265 270

Asn Lys Val Ile Glu Gln Leu Gly Thr Pro Cys Pro Glu Phe Met Lys

275 280 285

Lys Leu Gln Pro Thr Val Arg Asn Tyr Val Glu Asn Arg Pro Lys Tyr

290 295 300

Ala Gly Leu Thr Phe Pro Lys Leu Phe Pro Asp Ser Leu Phe Pro Ala

305 310 315 320

Asp Ser Glu His Asn Lys Leu Lys Ala Ser Gln Ala Arg Asp Leu Leu

325 330 335

Ser Lys Met Leu Val Ile Asp Pro Ala Lys Arg Ile Ser Val Asp Asp

340 345 350

Ala Leu Gln His Pro Tyr Ile Asn Val Trp Tyr Asp Pro Ala Glu Val

355 360 365

Glu Ala Pro Pro Pro Gln Ile Tyr Asp Lys Gln Leu Asp Glu Arg Glu

370 375 380

His Thr Ile Glu Glu Trp Lys Glu Leu Ile Tyr Lys Glu Val Met Asn

385 390 395 400

Ser Glu Glu Lys Thr Lys Asn Gly Val Val Lys Gly Gln Pro Ser Pro

405 410 415

Ser Gly Ala Ala Val Asn Ser Ser Glu Ser Leu Pro Pro Ser Ser Ser

420 425 430

Val Asn Asp Ile Ser Ser Met Ser Thr Asp Gln Thr Leu Ala Ser Asp

435 440 445

Thr Asp Ser Ser Leu Glu Ala Ser Ala Gly Pro Leu Gly Cys Cys Arg

450 455 460

6

351

Mouse

6

Met Gly Asn Ala Ala Ala Ala Lys Lys Gly Ser Glu Gln Glu Ser Val

1 5 10 15

Lys Glu Phe Leu Ala Lys Ala Lys Glu Asp Phe Leu Lys Lys Trp Glu

20 25 30

Thr Pro Ser Gln Asn Thr Ala Gln Leu Asp Gln Phe Asp Arg Ile Lys

35 40 45

Thr Leu Gly Thr Gly Ser Phe Gly Arg Val Met Leu Val Lys His Lys

50 55 60

Glu Ser Gly Asn His Tyr Ala Met Lys Ile Leu Asp Lys Gln Lys Val

65 70 75 80

Val Lys Leu Lys Gln Ile Glu His Thr Leu Asn Glu Lys Arg Ile Leu

85 90 95

Gln Ala Val Asn Phe Pro Phe Leu Val Lys Leu Glu Phe Ser Phe Lys

100 105 110

Asp Asn Ser Asn Leu Tyr Met Val Met Glu Tyr Val Ala Gly Gly Glu

115 120 125

Met Phe Ser His Leu Arg Arg Ile Gly Arg Phe Ser Glu Pro His Ala

130 135 140

Arg Phe Tyr Ala Ala Gln Ile Val Leu Thr Phe Glu Tyr Leu His Ser

145 150 155 160

Leu Asp Leu Ile Tyr Arg Asp Leu Lys Pro Glu Asn Leu Leu Ile Asp

165 170 175

Gln Gln Gly Tyr Ile Gln Val Thr Asp Phe Gly Phe Ala Lys Arg Val

180 185 190

Lys Gly Arg Thr Trp Thr Leu Cys Gly Thr Pro Glu Tyr Leu Ala Pro

195 200 205

Glu Ile Ile Leu Ser Lys Gly Tyr Asn Lys Ala Val Asp Trp Trp Ala

210 215 220

Leu Gly Val Leu Ile Tyr Glu Met Ala Ala Gly Tyr Pro Pro Phe Phe

225 230 235 240

Ala Asp Gln Pro Ile Gln Ile Tyr Glu Lys Ile Val Ser Gly Lys Val

245 250 255

Arg Phe Pro Ser His Phe Ser Ser Asp Leu Lys Asp Leu Leu Arg Asn

260 265 270

Leu Leu Gln Val Asp Leu Thr Lys Arg Phe Gly Asn Leu Lys Asn Gly

275 280 285

Val Asn Asp Ile Lys Asn His Lys Trp Phe Ala Thr Thr Asp Trp Ile

290 295 300

Ala Ile Tyr Gln Arg Lys Val Glu Ala Pro Phe Ile Pro Lys Phe Lys

305 310 315 320

Gly Pro Gly Asp Thr Ser Asn Phe Asp Asp Tyr Glu Glu Glu Glu Ile

325 330 335

Arg Val Ser Ile Asn Glu Lys Cys Gly Lys Glu Phe Thr Glu Phe

340 345 350

7

1382

Human

7

Met Gly Thr Gly Gly Arg Arg Gly Ala Ala Ala Ala Pro Leu Leu Val

1 5 10 15

Ala Val Ala Ala Leu Leu Leu Gly Ala Ala Gly His Leu Tyr Pro Gly

20 25 30

Glu Val Cys Pro Gly Met Asp Ile Arg Asn Asn Leu Thr Arg Leu His

35 40 45

Glu Leu Glu Asn Cys Ser Val Ile Glu Gly His Leu Gln Ile Leu Leu

50 55 60

Met Phe Lys Thr Arg Pro Glu Asp Phe Arg Asp Leu Ser Phe Pro Lys

65 70 75 80

Leu Ile Met Ile Thr Asp Tyr Leu Leu Leu Phe Arg Val Tyr Gly Leu

85 90 95

Glu Ser Leu Lys Asp Leu Phe Pro Asn Leu Thr Val Ile Arg Gly Ser

100 105 110

Arg Leu Phe Phe Asn Tyr Ala Leu Val Ile Phe Glu Met Val His Leu

115 120 125

Lys Glu Leu Gly Leu Tyr Asn Leu Met Asn Ile Thr Arg Gly Ser Val

130 135 140

Arg Ile Glu Lys Asn Asn Glu Leu Cys Tyr Leu Ala Thr Ile Asp Trp

145 150 155 160

Ser Arg Ile Leu Asp Ser Val Glu Asp Asn His Ile Val Leu Asn Lys

165 170 175

Asp Asp Asn Glu Glu Cys Gly Asp Ile Cys Pro Gly Thr Ala Lys Gly

180 185 190

Lys Thr Asn Cys Pro Ala Thr Val Ile Asn Gly Gln Phe Val Glu Arg

195 200 205

Cys Trp Thr His Ser His Cys Gln Lys Val Cys Pro Thr Ile Cys Lys

210 215 220

Ser His Gly Cys Thr Ala Glu Gly Leu Cys Cys His Ser Glu Cys Leu

225 230 235 240

Gly Asn Cys Ser Gln Pro Asp Asp Pro Thr Lys Cys Val Ala Cys Arg

245 250 255

Asn Phe Tyr Leu Asp Gly Arg Cys Val Glu Thr Cys Pro Pro Pro Tyr

260 265 270

Tyr His Phe Gln Asp Trp Arg Cys Val Asn Phe Ser Phe Cys Gln Asp

275 280 285

Leu His His Lys Cys Lys Asn Ser Arg Arg Gln Gly Cys His Gln Tyr

290 295 300

Val Ile His Asn Asn Lys Cys Ile Pro Glu Cys Pro Ser Gly Tyr Thr

305 310 315 320

Met Asn Ser Ser Asn Leu Leu Cys Thr Pro Cys Leu Gly Pro Cys Pro

325 330 335

Lys Val Cys His Leu Leu Glu Gly Glu Lys Thr Ile Asp Ser Val Thr

340 345 350

Ser Ala Gln Glu Leu Arg Gly Cys Thr Val Ile Asn Gly Ser Leu Ile

355 360 365

Ile Asn Ile Arg Gly Gly Asn Asn Leu Ala Ala Glu Leu Glu Ala Asn

370 375 380

Leu Gly Leu Ile Glu Glu Ile Ser Gly Tyr Leu Lys Ile Arg Arg Ser

385 390 395 400

Tyr Ala Leu Val Ser Leu Ser Phe Phe Arg Lys Leu Arg Leu Ile Arg

405 410 415

Gly Glu Thr Leu Glu Ile Gly Asn Tyr Ser Phe Tyr Ala Leu Asp Asn

420 425 430

Gln Asn Leu Arg Gln Leu Trp Asp Trp Ser Lys His Asn Leu Thr Thr

435 440 445

Thr Gln Gly Lys Leu Phe Phe His Tyr Asn Pro Lys Leu Cys Leu Ser

450 455 460

Glu Ile His Lys Met Glu Glu Val Ser Gly Thr Lys Gly Arg Gln Glu

465 470 475 480

Arg Asn Asp Ile Ala Leu Lys Thr Asn Gly Asp Lys Ala Ser Cys Glu

485 490 495

Asn Glu Leu Leu Lys Phe Ser Tyr Ile Arg Thr Ser Phe Asp Lys Ile

500 505 510

Leu Leu Arg Trp Glu Pro Tyr Trp Pro Pro Asp Phe Arg Asp Leu Leu

515 520 525

Gly Phe Met Leu Phe Tyr Lys Glu Ala Pro Tyr Gln Asn Val Thr Glu

530 535 540

Phe Asp Gly Gln Asp Ala Cys Gly Ser Asn Ser Trp Thr Val Val Asp

545 550 555 560

Ile Asp Pro Pro Leu Arg Ser Asn Asp Pro Lys Ser Gln Asn His Pro

565 570 575

Gly Trp Leu Met Arg Gly Leu Lys Pro Trp Thr Gln Tyr Ala Ile Phe

580 585 590

Val Lys Thr Leu Val Thr Phe Ser Asp Glu Arg Arg Thr Tyr Gly Ala

595 600 605

Lys Ser Asp Ile Ile Tyr Val Gln Thr Asp Ala Thr Asn Pro Ser Val

610 615 620

Pro Leu Asp Pro Ile Ser Val Ser Asn Ser Ser Ser Gln Ile Ile Leu

625 630 635 640

Lys Trp Lys Pro Pro Ser Asp Pro Asn Gly Asn Ile Thr His Tyr Leu

645 650 655

Val Phe Trp Glu Arg Gln Ala Glu Asp Ser Glu Leu Phe Glu Leu Asp

660 665 670

Tyr Cys Leu Lys Gly Leu Lys Leu Pro Ser Arg Thr Trp Ser Pro Pro

675 680 685

Phe Glu Ser Glu Asp Ser Gln Lys His Asn Gln Ser Glu Tyr Glu Asp

690 695 700

Ser Ala Gly Glu Cys Cys Ser Cys Pro Lys Thr Asp Ser Gln Ile Leu

705 710 715 720

Lys Glu Leu Glu Glu Ser Ser Phe Arg Lys Thr Phe Glu Asp Tyr Leu

725 730 735

His Asn Val Val Phe Val Pro Arg Lys Thr Ser Ser Gly Thr Gly Ala

740 745 750

Glu Asp Pro Arg Pro Ser Arg Lys Arg Arg Ser Leu Gly Asp Val Gly

755 760 765

Asn Val Thr Val Ala Val Pro Thr Val Ala Ala Phe Pro Asn Thr Ser

770 775 780

Ser Thr Ser Val Pro Thr Ser Pro Glu Glu His Arg Pro Phe Glu Lys

785 790 795 800

Val Val Asn Lys Glu Ser Leu Val Ile Ser Gly Leu Arg His Phe Thr

805 810 815

Gly Tyr Arg Ile Glu Leu Gln Ala Cys Asn Gln Asp Thr Pro Glu Glu

820 825 830

Arg Cys Ser Val Ala Ala Tyr Val Ser Ala Arg Thr Met Pro Glu Ala

835 840 845

Lys Ala Asp Asp Ile Val Gly Pro Val Thr His Glu Ile Phe Glu Asn

850 855 860

Asn Val Val His Leu Met Trp Gln Glu Pro Lys Glu Pro Asn Gly Leu

865 870 875 880

Ile Val Leu Tyr Glu Val Ser Tyr Arg Arg Tyr Gly Asp Glu Glu Leu

885 890 895

His Leu Cys Val Ser Arg Lys His Phe Ala Leu Glu Arg Gly Cys Arg

900 905 910

Leu Arg Gly Leu Ser Pro Gly Asn Tyr Ser Val Arg Ile Arg Ala Thr

915 920 925

Ser Leu Ala Gly Asn Gly Ser Trp Thr Glu Pro Thr Tyr Phe Tyr Val

930 935 940

Thr Asp Tyr Leu Asp Val Pro Ser Asn Ile Ala Lys Ile Ile Ile Gly

945 950 955 960

Pro Leu Ile Phe Val Phe Leu Phe Ser Val Val Ile Gly Ser Ile Tyr

965 970 975

Leu Phe Leu Arg Lys Arg Gln Pro Asp Gly Pro Leu Gly Pro Leu Tyr

980 985 990

Ala Ser Ser Asn Pro Glu Tyr Leu Ser Ala Ser Asp Val Phe Pro Cys

995 1000 1005

Ser Val Tyr Val Pro Asp Glu Trp Glu Val Ser Arg Glu Lys Ile

1010 1015 1020

Thr Leu Leu Arg Glu Leu Gly Gln Gly Ser Phe Gly Met Val Tyr

1025 1030 1035

Glu Gly Asn Ala Arg Asp Ile Ile Lys Gly Glu Ala Glu Thr Arg

1040 1045 1050

Val Ala Val Lys Thr Val Asn Glu Ser Ala Ser Leu Arg Glu Arg

1055 1060 1065

Ile Glu Phe Leu Asn Glu Ala Ser Val Met Lys Gly Phe Thr Cys

1070 1075 1080

His His Val Val Arg Leu Leu Gly Val Val Ser Lys Gly Gln Pro

1085 1090 1095

Thr Leu Val Val Met Glu Leu Met Ala His Gly Asp Leu Lys Ser

1100 1105 1110

Tyr Leu Arg Ser Leu Arg Pro Glu Ala Glu Asn Asn Pro Gly Arg

1115 1120 1125

Pro Pro Pro Thr Leu Gln Glu Met Ile Gln Met Ala Ala Glu Ile

1130 1135 1140

Ala Asp Gly Met Ala Tyr Leu Asn Ala Lys Lys Phe Val His Arg

1145 1150 1155

Asp Leu Ala Ala Arg Asn Cys Met Val Ala His Asp Phe Thr Val

1160 1165 1170

Lys Ile Gly Asp Phe Gly Met Thr Arg Asp Ile Tyr Glu Thr Asp

1175 1180 1185

Tyr Tyr Arg Lys Gly Gly Lys Gly Leu Leu Pro Val Arg Trp Met

1190 1195 1200

Ala Pro Glu Ser Leu Lys Asp Gly Val Phe Thr Thr Ser Ser Asp

1205 1210 1215

Met Trp Ser Phe Gly Val Val Leu Trp Glu Ile Thr Ser Leu Ala

1220 1225 1230

Glu Gln Pro Tyr Gln Gly Leu Ser Asn Glu Gln Val Leu Lys Phe

1235 1240 1245

Val Met Asp Gly Gly Tyr Leu Asp Gln Pro Asp Asn Cys Pro Glu

1250 1255 1260

Arg Val Thr Asp Leu Met Arg Met Cys Trp Gln Phe Asn Pro Lys

1265 1270 1275

Met Arg Pro Thr Phe Leu Glu Ile Val Asn Leu Leu Lys Asp Asp

1280 1285 1290

Leu His Pro Ser Phe Pro Glu Val Ser Phe Phe His Ser Glu Glu

1295 1300 1305

Asn Lys Ala Pro Glu Ser Glu Glu Leu Glu Met Glu Phe Glu Asp

1310 1315 1320

Met Glu Asn Val Pro Leu Asp Arg Ser Ser His Cys Gln Arg Glu

1325 1330 1335

Glu Ala Gly Gly Arg Asp Gly Gly Ser Ser Leu Gly Phe Lys Arg

1340 1345 1350

Ser Tyr Glu Glu His Ile Pro Tyr Thr His Met Asn Gly Gly Lys

1355 1360 1365

Lys Asn Gly Arg Ile Leu Thr Leu Pro Arg Ser Asn Pro Ser

1370 1375 1380

8

509

Human

8

Met Gly Cys Gly Cys Ser Ser His Pro Glu Asp Asp Trp Met Glu Asn

1 5 10 15

Ile Asp Val Cys Glu Asn Cys His Tyr Pro Ile Val Pro Leu Asp Gly

20 25 30

Lys Gly Thr Leu Leu Ile Arg Asn Gly Ser Glu Val Arg Asp Pro Leu

35 40 45

Val Thr Tyr Glu Gly Ser Asn Pro Pro Ala Ser Pro Leu Gln Asp Asn

50 55 60

Leu Val Ile Ala Leu His Ser Tyr Glu Pro Ser His Asp Gly Asp Leu

65 70 75 80

Gly Phe Glu Lys Gly Glu Gln Leu Arg Ile Leu Glu Gln Ser Gly Glu

85 90 95

Trp Trp Lys Ala Gln Ser Leu Thr Thr Gly Gln Glu Gly Phe Ile Pro

100 105 110

Phe Asn Phe Val Ala Lys Ala Asn Ser Leu Glu Pro Glu Pro Trp Phe

115 120 125

Phe Lys Asn Leu Ser Arg Lys Asp Ala Glu Arg Gln Leu Leu Ala Pro

130 135 140

Gly Asn Thr His Gly Ser Phe Leu Ile Arg Glu Ser Glu Ser Thr Ala

145 150 155 160

Gly Ser Phe Ser Leu Ser Val Arg Asp Phe Asp Gln Asn Gln Gly Glu

165 170 175

Val Val Lys His Tyr Lys Ile Arg Asn Leu Asp Asn Gly Gly Phe Tyr

180 185 190

Ile Ser Pro Arg Ile Thr Phe Pro Gly Leu His Glu Leu Val Arg His

195 200 205

Tyr Thr Asn Ala Ser Asp Gly Leu Cys Thr Arg Leu Ser Arg Pro Cys

210 215 220

Gln Thr Gln Lys Pro Gln Lys Pro Trp Trp Glu Asp Glu Trp Glu Val

225 230 235 240

Pro Arg Glu Thr Leu Lys Leu Val Glu Arg Leu Gly Ala Gly Gln Phe

245 250 255

Gly Glu Val Trp Met Gly Tyr Tyr Asn Gly His Thr Lys Val Ala Val

260 265 270

Lys Ser Leu Lys Gln Gly Ser Met Ser Pro Asp Ala Phe Leu Ala Glu

275 280 285

Ala Asn Leu Met Lys Gln Leu Gln His Gln Arg Leu Val Arg Leu Tyr

290 295 300

Ala Val Val Thr Gln Glu Pro Ile Tyr Ile Ile Thr Glu Tyr Met Glu

305 310 315 320

Asn Gly Ser Leu Val Asp Phe Leu Lys Thr Pro Ser Gly Ile Lys Leu

325 330 335

Thr Ile Asn Lys Leu Leu Asp Met Ala Ala Gln Ile Ala Glu Gly Met

340 345 350

Ala Phe Ile Glu Glu Arg Asn Tyr Ile His Arg Asp Leu Arg Ala Ala

355 360 365

Asn Ile Leu Val Ser Asp Thr Leu Ser Cys Lys Ile Ala Asp Phe Gly

370 375 380

Leu Ala Arg Leu Ile Glu Asp Asn Glu Tyr Thr Ala Arg Glu Gly Ala

385 390 395 400

Lys Phe Pro Ile Lys Trp Thr Ala Pro Glu Ala Ile Asn Tyr Gly Thr

405 410 415

Phe Thr Ile Lys Ser Asp Val Trp Ser Phe Gly Ile Leu Leu Thr Glu

420 425 430

Ile Val Thr His Gly Arg Ile Pro Tyr Pro Gly Met Thr Asn Pro Glu

435 440 445

Val Ile Gln Asn Leu Glu Arg Gly Tyr Arg Met Val Arg Pro Asp Asn

450 455 460

Cys Pro Glu Glu Leu Tyr Gln Leu Met Arg Leu Cys Trp Lys Glu Arg

465 470 475 480

Pro Glu Asp Arg Pro Thr Phe Asp Tyr Leu Arg Ser Val Leu Glu Asp

485 490 495

Phe Phe Thr Ala Thr Glu Gly Gln Tyr Gln Pro Gln Pro

500 505

9

619

Human

9

Met Pro Asp Pro Ala Ala His Leu Pro Phe Phe Tyr Gly Ser Ile Ser

1 5 10 15

Arg Ala Glu Ala Glu Glu His Leu Lys Leu Ala Gly Met Ala Asp Gly

20 25 30

Leu Phe Leu Leu Arg Gln Cys Leu Arg Ser Leu Gly Gly Tyr Val Leu

35 40 45

Ser Leu Val His Asp Val Arg Phe His His Phe Pro Ile Glu Arg Gln

50 55 60

Leu Asn Gly Thr Tyr Ala Ile Ala Gly Gly Lys Ala His Cys Gly Pro

65 70 75 80

Ala Glu Leu Cys Glu Phe Tyr Ser Arg Asp Pro Asp Gly Leu Pro Cys

85 90 95

Asn Leu Arg Lys Pro Cys Asn Arg Pro Ser Gly Leu Glu Pro Gln Pro

100 105 110

Gly Val Phe Asp Cys Leu Arg Asp Ala Met Val Arg Asp Tyr Val Arg

115 120 125

Gln Thr Trp Lys Leu Glu Gly Glu Ala Leu Glu Gln Ala Ile Ile Ser

130 135 140

Gln Ala Pro Gln Val Glu Lys Leu Ile Ala Thr Thr Ala His Glu Arg

145 150 155 160

Met Pro Trp Tyr His Ser Ser Leu Thr Arg Glu Glu Ala Glu Arg Lys

165 170 175

Leu Tyr Ser Gly Ala Gln Thr Asp Gly Lys Phe Leu Leu Arg Pro Arg

180 185 190

Lys Glu Gln Gly Thr Tyr Ala Leu Ser Leu Ile Tyr Gly Lys Thr Val

195 200 205

Tyr His Tyr Leu Ile Ser Gln Asp Lys Ala Gly Lys Tyr Cys Ile Pro

210 215 220

Glu Gly Thr Lys Phe Asp Thr Leu Trp Gln Leu Val Glu Tyr Leu Lys

225 230 235 240

Leu Lys Ala Asp Gly Leu Ile Tyr Cys Leu Lys Glu Ala Cys Pro Asn

245 250 255

Ser Ser Ala Ser Asn Ala Ser Gly Ala Ala Ala Pro Thr Leu Pro Ala

260 265 270

His Pro Ser Thr Leu Thr His Pro Gln Arg Arg Ile Asp Thr Leu Asn

275 280 285

Ser Asp Gly Tyr Thr Pro Glu Pro Ala Arg Ile Thr Ser Pro Asp Lys

290 295 300

Pro Arg Pro Met Pro Met Asp Thr Ser Val Tyr Glu Ser Pro Tyr Ser

305 310 315 320

Asp Pro Glu Glu Leu Lys Asp Lys Lys Leu Phe Leu Lys Arg Asp Asn

325 330 335

Leu Leu Ile Ala Asp Ile Glu Leu Gly Cys Gly Asn Phe Gly Ser Val

340 345 350

Arg Gln Gly Val Tyr Arg Met Arg Lys Lys Gln Ile Asp Val Ala Ile

355 360 365

Lys Val Leu Lys Gln Gly Thr Glu Lys Ala Asp Thr Glu Glu Met Met

370 375 380

Arg Glu Ala Gln Ile Met His Gln Leu Asp Asn Pro Tyr Ile Val Arg

385 390 395 400

Leu Ile Gly Val Cys Gln Ala Glu Ala Leu Met Leu Val Met Glu Met

405 410 415

Ala Gly Gly Gly Pro Leu His Lys Phe Leu Val Gly Lys Arg Glu Glu

420 425 430

Ile Pro Val Ser Asn Val Ala Glu Leu Leu His Gln Val Ser Met Gly

435 440 445

Met Lys Tyr Leu Glu Glu Lys Asn Phe Val His Arg Asp Leu Ala Ala

450 455 460

Arg Asn Val Leu Leu Val Asn Arg His Tyr Ala Lys Ile Ser Asp Phe

465 470 475 480

Gly Leu Ser Lys Ala Leu Gly Ala Asp Asp Ser Tyr Tyr Thr Ala Arg

485 490 495

Ser Ala Gly Lys Trp Pro Leu Lys Trp Tyr Ala Pro Glu Cys Ile Asn

500 505 510

Phe Arg Lys Phe Ser Ser Arg Ser Asp Val Trp Ser Tyr Gly Val Thr

515 520 525

Met Trp Glu Ala Leu Ser Tyr Gly Gln Lys Pro Tyr Lys Lys Met Lys

530 535 540

Gly Pro Glu Val Met Ala Phe Ile Glu Gln Gly Lys Arg Met Glu Cys

545 550 555 560

Pro Pro Glu Cys Pro Pro Glu Leu Tyr Ala Leu Met Ser Asp Cys Trp

565 570 575

Ile Tyr Lys Trp Glu Asp Arg Pro Asp Phe Leu Thr Val Glu Gln Arg

580 585 590

Met Arg Ala Cys Tyr Tyr Ser Leu Ala Ser Lys Val Glu Gly Pro Pro

595 600 605

Gly Ser Thr Gln Lys Ala Glu Ala Ala Cys Ala

610 615

10

416

Human

10

Leu Phe His Lys Ala Lys Phe Gln Glu Cys His Lys Asn Leu Ile Ser

1 5 10 15

Ile Val His Tyr Val Ala Phe Lys Val Leu Thr Trp Lys Arg Thr Lys

20 25 30

Ile Ser Asp Phe Asn Phe Tyr Gly Ser Leu Gly Ser Gly Ser Phe Gly

35 40 45

Thr Ala Lys Leu Cys Arg His Arg Gly Ser Gly Leu Phe Phe Cys Ser

50 55 60

Lys Thr Leu Arg Arg Glu Thr Ile Val His Glu Lys His Lys Glu His

65 70 75 80

Val Asn Asn Glu Ile Asn Ile Met Leu Asn Ile Ser His Pro Tyr Ile

85 90 95

Val Lys Thr Tyr Ser Thr Phe Asn Thr Pro Thr Lys Ile His Phe Ile

100 105 110

Met Glu Tyr Ala Gly Lys Lys Asp Leu Phe His His Leu Arg Ala Asn

115 120 125

Lys Cys Phe Thr Glu Gln Thr Thr Lys Leu Ile Val Ala Glu Ile Val

130 135 140

Leu Ala Ile Glu Tyr Leu His Ala Glu Asn Ile Ile Tyr Arg Asp Leu

145 150 155 160

Lys Pro Glu Asn Ile Leu Ile Asp Glu Lys Gly His Ile Lys Leu Thr

165 170 175

Asp Phe Gly Phe Ser Lys Lys Thr Val Gly Gly Lys Asn Thr Ser Ser

180 185 190

Val Cys Gly Thr Phe Asp Tyr Met Ala Pro Glu Ile Leu Asn Ser Ser

195 200 205

Asn Gly His Gly Lys Pro Val Asp Trp Trp Ala Leu Gly Val Val Val

210 215 220

Tyr Glu Leu Val Thr Gly Lys Leu Pro Phe Ser Asn Ser Lys Glu Ser

225 230 235 240

Leu Leu Asn Arg Lys Ala Asp Phe Gln Leu Ile Phe Gln Asn Ser Tyr

245 250 255

Leu Ser Asp Glu Ile Lys Asp Phe Ile Phe Gln Leu Leu Ser Val Asp

260 265 270

Pro Ser Lys Arg Leu Gly Thr Phe Asp Ser Cys Ser Ile Arg Asn His

275 280 285

Lys Trp Phe Ser Asp Ile Asn Trp Leu His Leu Glu Ser Lys Tyr Gln

290 295 300

Ile Asp Gly Pro Leu Ser Thr Leu Asn Ser Phe Ile Asn Cys Asp Phe

305 310 315 320

Asn Ile Asn Leu Leu Lys Lys Ser Lys Ser Tyr Thr Glu Gln Gln Gln

325 330 335

Gln Gln Gln Gln Leu Pro Gln Gln Gln Gln Gln Gln Gln Gln Asn Asn

340 345 350

Gln Leu Phe Asn Gln Thr Leu Gln Gln Gln Asn Phe Asn Phe His Pro

355 360 365

Ile Gln Pro Gln Gln Gln Gln Gln Gln Gln Phe Phe Asn Phe Gln Phe

370 375 380

Asn Asn Asn Asn Phe Asn Asn Asn Asn Asn Asn Asn Asn Asn Phe Asn

385 390 395 400

Glu Ala Cys Thr Ser Asn Thr Cys Gly Gly Thr Thr Ala Ser Ile Phe

405 410 415

11

1151

Yeast

11

Met Ser Phe Ser Gln Leu Glu Gln Asn Ile Lys Lys Lys Ile Ala Val

1 5 10 15

Glu Glu Asn Ile Ile Arg Gly Ala Ser Ala Leu Lys Lys Lys Thr Ser

20 25 30

Asn Val Met Val Ile Gln Lys Cys Asn Thr Asn Ile Arg Glu Ala Arg

35 40 45

Gln Asn Leu Glu Tyr Leu Glu Asp Ser Leu Lys Lys Leu Arg Leu Lys

50 55 60

Thr Ala Gln Gln Ser Gln Gly Glu Asn Gly Ser Glu Asp Asn Glu Arg

65 70 75 80

Phe Asn Ser Lys Glu Tyr Gly Phe Leu Ser Thr Lys Ser Pro Asn Glu

85 90 95

His Ile Phe Ser Arg Leu Asp Leu Val Lys Tyr Asp Cys Pro Ser Leu

100 105 110

Ala Gln Arg Ile Gln Tyr Met Leu Gln Gln Leu Glu Phe Lys Leu Gln

115 120 125

Val Glu Lys Gln Tyr Gln Glu Ala Asn Thr Lys Leu Thr Lys Leu Tyr

130 135 140

Gln Ile Asp Gly Asp Gln Arg Ser Ser Ser Ala Ala Glu Gly Gly Ala

145 150 155 160

Met Glu Ser Lys Tyr Arg Ile Gln Met Leu Asn Lys Ala Leu Lys Lys

165 170 175

Tyr Gln Ala Ile Asn Val Asp Phe Asp Gln Phe Lys His Gln Pro Asn

180 185 190

Asp Ile Met Asp Asn Gln Gln Pro Lys Phe Arg Arg Lys Gln Leu Thr

195 200 205

Gly Val Leu Thr Ile Gly Ile Thr Ala Ala Arg Asp Val Asp His Ile

210 215 220

Gln Ser Pro Met Phe Ala Arg Lys Pro Glu Ser Tyr Val Thr Ile Lys

225 230 235 240

Ile Asp Asp Thr Ile Lys Ala Arg Thr Lys Pro Ser Arg Asn Asp Arg

245 250 255

Trp Ser Glu Asp Phe Gln Ile Pro Val Glu Lys Gly Asn Glu Ile Glu

260 265 270

Ile Thr Val Tyr Asp Lys Val Asn Asp Ser Leu Ile Pro Val Ala Ile

275 280 285

Met Trp Leu Leu Leu Ser Asp Ile Ala Glu Glu Ile Arg Lys Lys Lys

290 295 300

Ala Gly Gln Thr Asn Glu Gln Gln Gly Trp Val Asn Ala Ser Asn Ile

305 310 315 320

Asn Gly Gly Ser Ser Leu Ala Ser Glu Glu Gly Ser Thr Leu Thr Ser

325 330 335

Thr Tyr Ser Asn Ser Ala Ile Gln Ser Thr Ser Ala Lys Asn Val Gln

340 345 350

Gly Glu Asn Thr Ser Thr Ser Gln Ile Ser Thr Asn Ser Trp Phe Val

355 360 365

Leu Glu Pro Ser Gly Gln Ile Leu Leu Thr Leu Gly Phe His Lys Ser

370 375 380

Ser Gln Ile Glu Arg Lys Gln Leu Met Gly Gly Leu His Arg His Gly

385 390 395 400

Ala Ile Ile Asn Arg Lys Glu Glu Ile Phe Glu Gln His Gly His His

405 410 415

Phe Val Gln Lys Ser Phe Tyr Asn Ile Met Cys Cys Ala Tyr Cys Gly

420 425 430

Asp Phe Leu Arg Tyr Thr Gly Phe Gln Cys Gln Asp Cys Lys Phe Leu

435 440 445

Cys His Lys Lys Cys Tyr Thr Asn Val Val Thr Lys Cys Ile Ala Lys

450 455 460

Thr Ser Thr Asp Thr Asp Pro Asp Glu Ala Lys Leu Asn His Arg Ile

465 470 475 480

Pro His Arg Phe Leu Pro Thr Ser Asn Arg Gly Thr Lys Trp Cys Cys

485 490 495

His Cys Gly Tyr Ile Leu Pro Trp Gly Arg His Lys Val Arg Lys Cys

500 505 510

Ser Glu Cys Gly Ile Met Cys His Ala Gln Cys Ala His Leu Val Pro

515 520 525

Asp Phe Cys Gly Met Ser Met Glu Met Ala Asn Lys Ile Leu Lys Thr

530 535 540

Ile Gln Asp Thr Lys Arg Asn Gln Glu Lys Lys Lys Arg Thr Val Pro

545 550 555 560

Ser Ala Gln Leu Gly Ser Ser Ile Gly Thr Ala Asn Gly Ser Asp Leu

565 570 575

Ser Pro Ser Lys Leu Ala Glu Arg Ala Asn Ala Pro Leu Pro Pro Gln

580 585 590

Pro Arg Lys His Asp Lys Thr Pro Ser Pro Gln Lys Val Gly Arg Asp

595 600 605

Ser Pro Thr Lys Gln His Asp Pro Ile Ile Asp Lys Lys Ile Ser Leu

610 615 620

Gln Thr His Gly Arg Glu Lys Leu Asn Lys Phe Ile Asp Glu Asn Glu

625 630 635 640

Ala Tyr Leu Asn Phe Thr Glu Gly Ala Gln Gln Thr Ala Glu Phe Ser

645 650 655

Ser Pro Glu Lys Thr Leu Asp Pro Thr Ser Asn Arg Arg Ser Leu Gly

660 665 670

Leu Thr Asp Leu Ser Ile Glu His Ser Gln Thr Trp Glu Ser Lys Asp

675 680 685

Asp Leu Met Arg Asp Glu Leu Glu Leu Trp Lys Ala Gln Arg Glu Glu

690 695 700

Met Glu Leu Glu Ile Lys Gln Asp Ser Gly Glu Ile Gln Glu Asp Leu

705 710 715 720

Glu Val Asp His Ile Asp Leu Glu Thr Lys Gln Lys Leu Asp Trp Glu

725 730 735

Asn Lys Asn Asp Phe Arg Glu Ala Asp Leu Thr Ile Asp Ser Thr His

740 745 750

Thr Asn Pro Phe Arg Asp Met Asn Ser Glu Thr Phe Gln Ile Glu Gln

755 760 765

Asp His Ala Ser Lys Glu Val Leu Gln Glu Thr Val Ser Leu Ala Pro

770 775 780

Thr Ser Thr His Pro Ser Arg Thr Thr Asp Gln Gln Ser Pro Gln Lys

785 790 795 800

Ser Gln Thr Ser Thr Ser Ala Lys His Lys Lys Arg Ala Ala Lys Arg

805 810 815

Arg Lys Val Ser Leu Asp Asn Phe Val Leu Leu Lys Val Leu Gly Lys

820 825 830

Gly Asn Phe Gly Lys Val Ile Leu Ser Lys Ser Lys Asn Thr Asp Arg

835 840 845

Leu Cys Ala Ile Lys Val Leu Lys Lys Asp Asn Ile Ile Gln Asn His

850 855 860

Asp Ile Glu Ser Ala Arg Ala Glu Lys Lys Val Phe Leu Leu Ala Thr

865 870 875 880

Lys Thr Lys His Pro Phe Leu Thr Asn Leu Tyr Cys Ser Phe Gln Thr

885 890 895

Glu Asn Arg Ile Tyr Phe Ala Met Glu Phe Ile Gly Gly Gly Asp Leu

900 905 910

Met Trp His Val Gln Asn Gln Arg Leu Ser Val Arg Arg Ala Lys Phe

915 920 925

Tyr Ala Ala Glu Val Leu Leu Ala Leu Lys Tyr Phe His Asp Asn Gly

930 935 940

Val Ile Tyr Arg Asp Leu Lys Leu Glu Asn Ile Leu Leu Thr Pro Glu

945 950 955 960

Gly His Ile Lys Ile Ala Asp Tyr Gly Leu Cys Lys Asp Glu Met Trp

965 970 975

Tyr Gly Asn Arg Thr Ser Thr Phe Cys Gly Thr Pro Glu Phe Met Ala

980 985 990

Pro Glu Ile Leu Lys Glu Gln Glu Tyr Thr Lys Ala Val Asp Trp Trp

995 1000 1005

Ala Phe Gly Val Leu Leu Tyr Gln Met Leu Leu Cys Gln Ser Pro

1010 1015 1020

Phe Ser Gly Asp Asp Glu Asp Glu Val Phe Asn Ala Ile Leu Thr

1025 1030 1035

Asp Glu Pro Leu Tyr Pro Ile Asp Met Ala Gly Glu Ile Val Gln

1040 1045 1050

Ile Phe Gln Gly Leu Leu Thr Lys Asp Pro Glu Lys Arg Leu Gly

1055 1060 1065

Ala Gly Pro Arg Asp Ala Asp Glu Val Met Glu Glu Pro Phe Phe

1070 1075 1080

Arg Asn Ile Asn Phe Asp Asp Ile Leu Asn Leu Arg Val Lys Pro

1085 1090 1095

Pro Tyr Ile Pro Glu Ile Lys Ser Pro Glu Asp Thr Ser Tyr Phe

1100 1105 1110

Glu Gln Glu Phe Thr Ser Ala Pro Pro Thr Leu Thr Pro Leu Pro

1115 1120 1125

Ser Val Leu Thr Thr Ser Gln Gln Glu Glu Phe Arg Gly Phe Ser

1130 1135 1140

Phe Met Pro Asp Asp Leu Asp Leu

1145 1150

12

484

Human

12

Met Arg His Ser Lys Arg Thr Tyr Cys Pro Asp Trp Asp Asp Lys Asp

1 5 10 15

Trp Asp Tyr Gly Lys Trp Arg Ser Ser Ser Ser His Lys Arg Arg Lys

20 25 30

Arg Ser His Ser Ser Ala Gln Glu Asn Lys Arg Cys Lys Tyr Asn His

35 40 45

Ser Lys Met Cys Asp Ser His Tyr Leu Glu Ser Arg Ser Ile Asn Glu

50 55 60

Lys Asp Tyr His Ser Arg Arg Tyr Ile Asp Glu Tyr Arg Asn Asp Tyr

65 70 75 80

Thr Gln Gly Cys Glu Pro Gly His Arg Gln Arg Asp His Glu Ser Arg

85 90 95

Tyr Gln Asn His Ser Ser Lys Ser Ser Gly Arg Ser Gly Arg Ser Ser

100 105 110

Tyr Lys Ser Lys His Arg Ile His His Ser Thr Ser His Arg Arg Ser

115 120 125

His Gly Lys Ser His Arg Arg Lys Arg Thr Arg Ser Val Glu Asp Asp

130 135 140

Glu Glu Gly His Leu Ile Cys Gln Ser Gly Asp Val Leu Ser Ala Arg

145 150 155 160

Tyr Glu Ile Val Asp Thr Leu Gly Glu Gly Ala Phe Gly Lys Val Val

165 170 175

Glu Cys Ile Asp His Lys Ala Gly Gly Arg His Val Ala Val Lys Ile

180 185 190

Val Lys Asn Val Asp Arg Tyr Cys Glu Ala Ala Arg Ser Glu Ile Gln

195 200 205

Val Leu Glu His Leu Asn Thr Thr Asp Pro Asn Ser Thr Phe Arg Cys

210 215 220

Val Gln Met Leu Glu Trp Phe Glu His His Gly His Ile Cys Ile Val

225 230 235 240

Phe Glu Leu Leu Gly Leu Ser Thr Tyr Asp Phe Ile Lys Glu Asn Gly

245 250 255

Phe Leu Pro Phe Arg Leu Asp His Ile Arg Lys Met Ala Tyr Gln Ile

260 265 270

Cys Lys Ser Val Asn Phe Leu His Ser Asn Lys Leu Thr His Thr Asp

275 280 285

Leu Lys Pro Glu Asn Ile Leu Phe Val Gln Ser Asp Tyr Thr Glu Ala

290 295 300

Tyr Asn Pro Lys Ile Lys Arg Asp Glu Arg Thr Leu Ile Asn Pro Asp

305 310 315 320

Ile Lys Val Val Asp Phe Gly Ser Ala Thr Tyr Asp Asp Glu His His

325 330 335

Ser Thr Leu Val Ser Thr Arg His Tyr Arg Ala Pro Glu Val Ile Leu

340 345 350

Ala Leu Gly Trp Ser Gln Pro Cys Asp Val Trp Ser Ile Gly Cys Ile

355 360 365

Leu Ile Glu Tyr Tyr Leu Gly Phe Thr Val Phe Pro Thr His Asp Ser

370 375 380

Lys Glu His Leu Ala Met Met Glu Arg Ile Leu Gly Pro Leu Pro Lys

385 390 395 400

His Met Ile Gln Lys Thr Arg Lys Arg Lys Tyr Phe His His Asp Arg

405 410 415

Leu Asp Trp Asp Glu His Ser Ser Ala Gly Arg Tyr Val Ser Arg Ala

420 425 430

Cys Lys Pro Leu Lys Glu Phe Met Leu Ser Gln Asp Val Glu His Glu

435 440 445

Arg Leu Phe Asp Leu Ile Gln Lys Met Leu Glu Tyr Asp Pro Ala Lys

450 455 460

Arg Ile Thr Leu Arg Glu Ala Leu Lys His Pro Phe Phe Asp Leu Leu

465 470 475 480

Lys Lys Ser Ile

13

499

Human

13

Met Pro His Pro Arg Arg Tyr His Ser Ser Glu Arg Gly Ser Arg Gly

1 5 10 15

Ser Tyr Arg Glu His Tyr Arg Ser Arg Lys His Lys Arg Arg Arg Ser

20 25 30

Arg Ser Trp Ser Ser Ser Ser Asp Arg Thr Arg Arg Arg Arg Arg Glu

35 40 45

Asp Ser Tyr His Val Arg Ser Arg Ser Ser Tyr Asp Asp Arg Ser Ser

50 55 60

Asp Arg Arg Val Tyr Asp Arg Arg Tyr Cys Gly Ser Tyr Arg Arg Asn

65 70 75 80

Asp Tyr Ser Arg Asp Arg Gly Asp Ala Tyr Tyr Asp Thr Asp Tyr Arg

85 90 95

His Ser Tyr Glu Tyr Gln Arg Glu Asn Ser Ser Tyr Arg Ser Gln Arg

100 105 110

Ser Ser Arg Arg Lys His Arg Arg Arg Arg Arg Arg Ser Arg Thr Phe

115 120 125

Ser Arg Ser Ser Ser Gln His Ser Ser Arg Arg Ala Lys Ser Val Glu

130 135 140

Asp Asp Ala Glu Gly His Leu Ile Tyr His Val Gly Asp Trp Leu Gln

145 150 155 160

Glu Arg Tyr Glu Ile Val Ser Thr Leu Gly Glu Gly Thr Phe Gly Arg

165 170 175

Val Val Gln Cys Val Asp His Arg Arg Gly Gly Ala Arg Val Ala Leu

180 185 190

Lys Ile Ile Lys Asn Val Glu Lys Tyr Lys Glu Ala Ala Arg Leu Glu

195 200 205

Ile Asn Val Leu Glu Lys Ile Asn Glu Lys Asp Pro Asp Asn Lys Asn

210 215 220

Leu Cys Val Gln Met Phe Asp Trp Phe Asp Tyr His Gly His Met Cys

225 230 235 240

Ile Ser Phe Glu Leu Leu Gly Leu Ser Thr Phe Asp Phe Leu Lys Asp

245 250 255

Asn Asn Tyr Leu Pro Tyr Pro Ile His Gln Val Arg His Met Ala Phe

260 265 270

Gln Leu Cys Gln Ala Val Lys Phe Leu His Asp Asn Lys Leu Thr His

275 280 285

Thr Asp Leu Lys Pro Glu Asn Ile Leu Phe Val Asn Ser Asp Tyr Glu

290 295 300

Leu Thr Tyr Asn Leu Glu Lys Lys Arg Asp Glu Arg Ser Val Lys Ser

305 310 315 320

Thr Ala Val Arg Val Val Asp Phe Gly Ser Ala Thr Phe Asp His Glu

325 330 335

His His Ser Thr Ile Val Ser Thr Arg His Tyr Arg Ala Pro Glu Val

340 345 350

Ile Leu Glu Leu Gly Trp Ser Gln Pro Cys Asp Val Trp Ser Ile Gly

355 360 365

Cys Ile Ile Phe Glu Tyr Tyr Val Gly Phe Thr Leu Phe Gln Thr His

370 375 380

Asp Asn Arg Glu His Leu Ala Met Met Glu Arg Ile Leu Gly Pro Ile

385 390 395 400

Pro Ser Arg Met Ile Arg Lys Thr Arg Lys Gln Lys Tyr Phe Tyr Arg

405 410 415

Gly Arg Leu Asp Trp Asp Glu Asn Thr Ser Ala Gly Arg Tyr Val Arg

420 425 430

Glu Asn Cys Lys Pro Leu Arg Arg Tyr Leu Thr Ser Glu Ala Glu Glu

435 440 445

His His Gln Leu Phe Asp Leu Ile Glu Ser Met Leu Glu Tyr Glu Pro

450 455 460

Ala Lys Arg Leu Thr Leu Gly Glu Ala Leu Gln His Pro Phe Phe Ala

465 470 475 480

Arg Leu Arg Ala Glu Pro Pro Asn Lys Leu Trp Asp Ser Ser Arg Asp

485 490 495

Ile Ser Arg

14

517

DROME - Drosophila Melanogaster Fruit Fly

14

Met Cys Val Arg Phe Gln Met Pro Arg Thr Arg Arg Leu His His Ser

1 5 10 15

Arg Asp Arg Ser Ser Ala Gly Thr Arg Asp Lys Arg Arg Arg His Asp

20 25 30

Thr Ala Asp His Ser Pro Pro Leu Ala Glu Ala Pro Ser Pro Pro Arg

35 40 45

Ile Thr Asn Thr His His Thr Arg Ser Ala Ala Lys Arg Arg Arg His

50 55 60

Glu Leu Asp Ala Lys Lys Ala Gln Ile Ser Lys Glu Pro Thr Phe Asp

65 70 75 80

Asp Ser Ile Ser Thr Arg Arg Arg Lys Glu Arg Ser Lys Arg Ser His

85 90 95

Arg Lys Ser Pro Ala Ala Ser Arg Arg Gln His Lys Tyr Arg Tyr Arg

100 105 110

Asp Glu Thr Ser His Ser Ser Ser Arg Arg Arg His Arg Asp Arg Ala

115 120 125

Lys Asp Glu Arg Asp Ser Gly Arg Asn Asn Arg Gln Ser Gln Ala Lys

130 135 140

Thr Ala Lys Pro Val Ile Gln Asp Asp Ala Asp Gly His Leu Ile Tyr

145 150 155 160

His Thr Gly Asp Ile Leu His His Arg Tyr Lys Ile Met Ala Thr Leu

165 170 175

Gly Glu Gly Thr Phe Gly Arg Val Val Lys Val Lys Asp Met Glu Arg

180 185 190

Asp Tyr Cys Met Ala Leu Lys Ile Ile Lys Asn Val Glu Lys Tyr Arg

195 200 205

Glu Ala Ala Lys Leu Glu Ile Asn Ala Leu Glu Lys Ile Ala Gln Lys

210 215 220

Asp Pro His Cys Asp His Leu Cys Val Lys Met Ile Asp Trp Phe Asp

225 230 235 240

Tyr His Gly His Met Cys Ile Val Phe Glu Met Leu Gly Leu Ser Val

245 250 255

Phe Asp Phe Leu Arg Glu Asn Asn Tyr Glu Pro Tyr Pro Leu Asp Gln

260 265 270

Val Arg His Met Ala Tyr Gln Leu Cys Tyr Ser Val Lys Phe Leu His

275 280 285

Asp Asn Arg Leu Thr His Thr Asp Leu Lys Pro Glu Asn Ile Leu Phe

290 295 300

Val Asp Ser Asp Tyr Thr Ser His Tyr Asn His Lys Ile Asn Arg Glu

305 310 315 320

Val Arg Arg Val Lys Asn Thr Asp Val Arg Leu Ile Asp Phe Gly Ser

325 330 335

Ala Thr Phe Asp His Glu His His Ser Thr Ile Val Ser Thr Arg His

340 345 350

Tyr Arg Ala Pro Glu Val Ile Leu Glu Leu Gly Trp Ser Gln Pro Cys

355 360 365

Asp Val Trp Ser Ile Gly Cys Ile Leu Phe Glu Leu Tyr Leu Gly Ile

370 375 380

Thr Leu Phe Gln Thr His Asp Asn Arg Glu His Leu Ala Met Met Glu

385 390 395 400

Arg Ile Leu Gly Gln Ile Pro Tyr Arg Met Ala Arg Lys Thr Lys Thr

405 410 415

Lys Tyr Phe Tyr His Gly Lys Leu Asp Trp Asp Glu Lys Ser Ser Ala

420 425 430

Gly Arg Tyr Val Arg Asp His Cys Lys Pro Leu Phe Leu Cys Gln Leu

435 440 445

Ser Asp Ser Glu Asp His Cys Glu Leu Phe Ser Leu Ile Lys Lys Met

450 455 460

Leu Glu Tyr Glu Pro Ser Ser Arg Ile Thr Leu Gly Glu Ala Leu His

465 470 475 480

His Pro Phe Phe Asp Arg Leu Pro Pro His His Arg Val Gly Glu Val

485 490 495

Ser Asn Lys Gln Pro Leu Ser Ser Gly Ser Ser Ser Arg Glu Arg Ser

500 505 510

His Ser Leu Ser Arg

515

15

544

Human

15

Met Gly Ser Asp Gly Ser Ser Leu Ser Pro Lys Val Ser Gln Pro Gly

1 5 10 15

His Thr Glu Ile Val Asp His Val Ser Glu Lys Val Ile Thr Asn Gly

20 25 30

Lys Asn Val Asn Lys Lys Val Asn Ser Glu Val Asp Gly Lys Ser Met

35 40 45

Val Glu Lys Val Lys Thr His Glu Glu Asn Ala Glu Asp Tyr His Tyr

50 55 60

Gly Gly Tyr His Pro Val Tyr Ile Gly Glu Glu Phe His His Arg Arg

65 70 75 80

Tyr Val Val Glu Arg Lys Leu Gly Trp Gly His Phe Ser Thr Val Trp

85 90 95

Leu Ala Tyr Asp Arg Ala Ala Lys Arg Arg Val Ala Leu Lys Val Val

100 105 110

Arg Ser Ala Glu His Tyr Arg Glu Thr Ser Ile Asp Glu Ile Arg Ile

115 120 125

Leu Gln Lys Ile Arg Glu Gly Asp Glu Lys His Leu Gly Lys Lys His

130 135 140

Ile Ile Ser Leu Leu Asp Tyr Phe Val His Arg Gly Pro Asn Gly Ala

145 150 155 160

His Val Cys Met Val Phe Glu Val Leu Gly Glu Asn Leu Leu Ser Leu

165 170 175

Ile Gln Ser Tyr Gly His Arg Gly Val Pro Val Gly Ile Val Lys Gln

180 185 190

Ile Ala Tyr Gln Leu Leu Ile Ala Leu Asp Tyr Leu His Arg Glu Cys

195 200 205

Gly Ile Ile His Thr Asp Leu Lys Pro Glu Asn Val Leu Ile Cys Ile

210 215 220

Asp Gln Asp Ala Leu Gln His Ile Glu Ala Pro Ala Thr Thr Ser Ser

225 230 235 240

Pro Thr Ser Asn Thr Ser Ser Ser Lys Thr Arg Asn Asn Thr Gly Tyr

245 250 255

Thr Ala Lys Ala Pro Ile Ile Lys Arg Gly Gln Ser Val Asp Asn Ser

260 265 270

Ala Gln Glu Arg Lys Thr Phe Ala Lys Asn Pro Thr Lys Asn Ser Lys

275 280 285

Pro Ala Gly Gln Val Ile Pro Ser Ser Pro Phe Thr Ser Thr Leu Ser

290 295 300

Arg Phe Pro Ser Leu Glu Gly Ala Val Ser Glu Ile Ser Leu Arg Asp

305 310 315 320

Ser Gln Lys His Asn Ser His Pro Asn Ser Pro Phe Ser Ser Gly Asp

325 330 335

Asn Ser Leu Ile Leu Asp Gly Val Asn Gly Ser Gln Glu Pro Val Pro

340 345 350

Lys Ile Thr Val Lys Ile Ala Asp Leu Gly Asn Ala Cys Trp Thr Arg

355 360 365

Lys His Phe Thr Asn Asp Val Gln Thr Arg Gln Tyr Arg Ser Pro Glu

370 375 380

Val Ile Leu Gly Cys Arg Trp Gly Ala Ser Ala Asp Cys Trp Ser Phe

385 390 395 400

Ala Cys Ile Ile Phe Glu Leu Leu Thr Gly Asp Tyr Leu Phe Asp Pro

405 410 415

Arg Asn Gly Asn Ser Tyr Ser Lys Glu Asp Asp His Ile Ala Gln Ile

420 425 430

Ile Glu Leu Leu Val Asn Tyr Pro Lys Gln Met Ala Leu Ser Gly Lys

435 440 445

His Ser Arg Asp Leu Phe Asn Arg Arg Gly Glu Leu Arg Asn Ile His

450 455 460

Lys Leu Lys Phe Trp Pro Leu Lys Asp Val Leu Glu Gln Lys Tyr His

465 470 475 480

Phe Ser Ala Glu Leu Ala Gln Gln Ile Ser Asp Phe Leu Ser Pro Met

485 490 495

Leu Cys Phe Asp Pro Ala Lys Arg Thr Asn Ala Gly Tyr Met Ser Asn

500 505 510

Ser Pro Trp Leu Arg Glu Val Ala Asp Pro Thr Phe Lys Ile Glu Thr

515 520 525

Thr Gly Ala Thr Gly Glu Asp Val Pro Gly Trp Ala Thr Glu Ile Arg

530 535 540

16

508

Yeast

16

Met Ala Ser Leu Phe Arg Pro Pro Glu Ser Ala Lys Cys Asn Pro Asn

1 5 10 15

Ser Pro Arg Leu Lys Leu Pro Leu Leu Arg Asn Asn Gln Val Asp Glu

20 25 30

Asn Asn Ile Tyr Leu Thr Ser Asn Gly Ser Ser Thr Thr Ala Tyr Ser

35 40 45

Ser His Thr Pro Glu Pro Leu Thr Ser Ser Thr Ser Thr Leu Phe Ser

50 55 60

Gln Thr Arg Leu His Pro Ser Asp Ser Ser Met Thr Leu Asn Thr Met

65 70 75 80

Lys Lys Arg Pro Ala Pro Pro Ser Leu Pro Ser Leu Ser Ile Asn Ser

85 90 95

Gln Ser Lys Cys Lys Thr Leu Pro Glu Leu Val Pro Ile Ala Asp Val

100 105 110

Ser Asp Gly Lys His Asp Leu Gly Leu Lys Gln Arg Val Ile Ala Glu

115 120 125

Asn Glu Leu Ser Gly Asn Ser Asp Leu Thr Pro Ser Ser Met Ala Ser

130 135 140

Pro Phe Ser His Thr Asn Thr Ser Ser Pro Tyr Leu Arg Asn Asp Leu

145 150 155 160

Ser Asn Ser Val Gly Ser Asp Phe Ser Asn Leu Ile Ser Ala Tyr Glu

165 170 175

Gln Ser Ser Ser Pro Ile Lys Ser Ser Ser Gln Pro Lys Ser Ser Ser

180 185 190

Glu Ser Tyr Ile Asp Leu Asn Ser Val Arg Asp Val Asp Gln Leu Asp

195 200 205

Glu Asn Gly Trp Lys Tyr Ala Asn Leu Lys Asp Arg Ile Glu Thr Leu

210 215 220

Gly Ile Leu Gly Glu Gly Ala Gly Gly Ser Val Ser Lys Cys Lys Leu

225 230 235 240

Lys Asn Gly Ser Lys Ile Phe Ala Leu Lys Val Ile Asn Thr Leu Asn

245 250 255

Thr Asp Pro Glu Tyr Gln Lys Gln Ile Phe Arg Glu Leu Gln Phe Asn

260 265 270

Arg Ser Phe Gln Ser Glu Tyr Ile Val Arg Tyr Tyr Gly Met Phe Thr

275 280 285

Asp Asp Glu Asn Ser Ser Ile Tyr Ile Ala Met Glu Tyr Met Gly Gly

290 295 300

Arg Ser Leu Asp Ala Ile Tyr Lys Asn Leu Leu Glu Arg Gly Gly Arg

305 310 315 320

Ile Ser Glu Lys Val Leu Gly Lys Ile Ala Glu Ala Val Leu Arg Gly

325 330 335

Leu Ser Tyr Leu His Glu Lys Lys Val Ile His Arg Asp Ile Lys Pro

340 345 350

Gln Asn Ile Leu Leu Asn Glu Asn Gly Gln Val Lys Leu Cys Asp Phe

355 360 365

Gly Val Ser Gly Glu Ala Val Asn Ser Leu Ala Thr Thr Phe Thr Gly

370 375 380

Thr Ser Phe Tyr Met Ala Pro Glu Arg Ile Gln Gly Gln Pro Tyr Ser

385 390 395 400

Val Thr Ser Asp Val Trp Ser Leu Gly Leu Thr Ile Leu Glu Val Ala

405 410 415

Asn Gly Lys Phe Pro Cys Ser Ser Glu Lys Met Ala Ala Asn Ile Ala

420 425 430

Pro Phe Glu Leu Leu Met Trp Ile Leu Thr Phe Thr Pro Glu Leu Lys

435 440 445

Asp Glu Pro Glu Ser Asn Ile Ile Trp Ser Pro Ser Phe Lys Ser Phe

450 455 460

Ile Asp Tyr Cys Leu Lys Lys Asp Ser Arg Glu Arg Pro Ser Pro Arg

465 470 475 480

Gln Met Ile Asn His Pro Trp Ile Lys Gly Gln Met Lys Lys Asn Val

485 490 495

Asn Met Glu Lys Phe Val Arg Lys Cys Trp Lys Asp

500 505

17

506

Yeast

17

Met Ala Ser Met Phe Arg Pro Pro Glu Ser Asn Arg Ser His Gln Lys

1 5 10 15

Thr Pro Lys Leu Thr Leu Pro Val Asn Leu Val Gln Asn Ala Lys Ser

20 25 30

Thr Asn Asp Gly Gln His Leu Asn Arg Ser Pro Tyr Ser Ser Val Asn

35 40 45

Glu Ser Pro Tyr Ser Asn Asn Ser Thr Ser Ala Thr Ser Thr Thr Ser

50 55 60

Ser Met Ala Ser Asn Ser Thr Leu Leu Tyr Asn Arg Ser Ser Thr Thr

65 70 75 80

Thr Ile Lys Asn Arg Pro Val Pro Pro Pro Leu Pro Pro Leu Val Leu

85 90 95

Thr Gln Lys Lys Asp Gly Ile Glu Tyr Arg Val Ala Gly Asp Ser Gln

100 105 110

Leu Ser Glu Arg Phe Ser Asn Leu His Val Asp Ile Thr Tyr Lys Glu

115 120 125

Leu Leu Ser Ser Ala Pro Ile Ser Thr Lys Leu Ser Asn Ile Asp Thr

130 135 140

Thr Phe Ile Lys Lys Asp Leu Asp Thr Pro Glu Gly Glu Asp Ser Tyr

145 150 155 160

Pro Ser Thr Leu Leu Ser Ala Tyr Asp Phe Ser Ser Ser Gly Ser Asn

165 170 175

Ser Ala Pro Leu Ser Ala Asn Asn Ile Ile Ser Cys Ser Asn Leu Ile

180 185 190

Gln Gly Lys Asp Val Asp Gln Leu Glu Glu Glu Ala Trp Arg Phe Gly

195 200 205

His Leu Lys Asp Glu Ile Thr Thr Leu Gly Ile Leu Gly Glu Gly Ala

210 215 220

Gly Gly Ser Val Ala Lys Cys Arg Leu Lys Asn Gly Lys Lys Val Phe

225 230 235 240

Ala Leu Lys Thr Ile Asn Thr Met Asn Thr Asp Pro Glu Tyr Gln Lys

245 250 255

Gln Ile Phe Arg Glu Leu Gln Phe Asn Lys Ser Phe Lys Ser Asp Tyr

260 265 270

Ile Val Gln Tyr Tyr Gly Met Phe Thr Asp Glu Gln Ser Ser Ser Ile

275 280 285

Tyr Ile Ala Met Glu Tyr Met Gly Gly Lys Ser Leu Glu Ala Thr Tyr

290 295 300

Lys Asn Leu Leu Lys Arg Gly Gly Arg Ile Ser Glu Arg Val Ile Gly

305 310 315 320

Lys Ile Ala Glu Ser Val Leu Arg Gly Leu Ser Tyr Leu His Glu Arg

325 330 335

Lys Val Ile His Arg Asp Ile Lys Pro Gln Asn Ile Leu Leu Asn Glu

340 345 350

Lys Gly Glu Ile Lys Leu Cys Asp Phe Gly Val Ser Gly Glu Ala Val

355 360 365

Asn Ser Leu Ala Met Thr Phe Thr Gly Thr Ser Phe Tyr Met Ala Pro

370 375 380

Glu Arg Ile Gln Gly Gln Pro Tyr Ser Val Thr Cys Asp Val Trp Ser

385 390 395 400

Leu Gly Leu Thr Leu Leu Glu Val Ala Gly Gly Arg Phe Pro Phe Glu

405 410 415

Ser Asp Lys Ile Thr Gln Asn Val Ala Pro Ile Glu Leu Leu Thr Met

420 425 430

Ile Leu Thr Phe Ser Pro Gln Leu Lys Asp Glu Pro Glu Leu Asp Ile

435 440 445

Ser Trp Ser Lys Thr Phe Arg Ser Phe Ile Asp Tyr Cys Leu Lys Lys

450 455 460

Asp Ala Arg Glu Arg Pro Ser Pro Arg Gln Met Leu Lys His Pro Trp

465 470 475 480

Ile Val Gly Gln Met Lys Lys Lys Val Asn Met Glu Arg Phe Val Lys

485 490 495

Lys Cys Trp Glu Lys Glu Lys Asp Gly Ile

500 505

18

699

EMENI - Aspergillus Nidulans

18

Met Ala Ile Ala Leu Ala Glu Ala Asp Lys Tyr Glu Val Leu Glu Lys

1 5 10 15

Ile Gly Cys Gly Ser Phe Gly Ile Ile Arg Lys Val Lys Arg Lys Ser

20 25 30

Asp Gly Phe Ile Leu Cys Arg Lys Glu Ile Asn Tyr Ile Lys Met Ser

35 40 45

Thr Lys Glu Arg Glu Gln Leu Thr Ala Glu Phe Asn Ile Leu Ser Ser

50 55 60

Leu Arg His Pro Asn Ile Val Ala Tyr Tyr His Arg Glu His Leu Lys

65 70 75 80

Ala Ser Gln Asp Leu Tyr Leu Tyr Met Glu Tyr Cys Gly Gly Gly Asp

85 90 95

Leu Ser Met Val Ile Lys Asn Leu Lys Arg Thr Asn Lys Tyr Ala Glu

100 105 110

Glu Asp Phe Val Trp Arg Ile Leu Ser Gln Leu Val Thr Ala Leu Tyr

115 120 125

Arg Cys His Tyr Gly Thr Asp Pro Ala Glu Val Gly Ser Asn Leu Leu

130 135 140

Gly Pro Ala Pro Lys Pro Ser Gly Leu Lys Gly Lys Gln Ala Gln Met

145 150 155 160

Thr Ile Leu His Arg Asp Leu Lys Pro Glu Asn Ile Phe Leu Gly Ser

165 170 175

Asp Asn Thr Val Lys Leu Gly Asp Phe Gly Leu Ser Lys Leu Met His

180 185 190

Ser His Asp Phe Ala Ser Thr Tyr Val Gly Thr Pro Phe Tyr Met Ser

195 200 205

Pro Glu Ile Cys Ala Ala Glu Lys Tyr Thr Leu Arg Ser Asp Ile Trp

210 215 220

Ala Val Gly Cys Ile Met Tyr Glu Leu Cys Gln Arg Glu Pro Pro Phe

225 230 235 240

Asn Ala Arg Thr His Ile Gln Leu Val Gln Lys Ile Arg Glu Gly Lys

245 250 255

Phe Ala Pro Leu Pro Asp Phe Tyr Ser Ser Glu Leu Lys Asn Val Ile

260 265 270

Ala Ser Cys Leu Arg Val Asn Pro Asp His Arg Pro Asp Thr Ala Thr

275 280 285

Leu Ile Asn Thr Pro Val Ile Arg Leu Met Arg Arg Glu Val Glu Leu

290 295 300

Asn Asn Leu Ser Arg Ala Ala Arg Lys Arg Glu Glu Ala Thr Met Gln

305 310 315 320

Lys Ala Lys Asp Val Glu Gln Ala Phe Ala Lys Leu Glu Lys Glu Lys

325 330 335

Gln Gln Ile Arg Ser Glu Leu Glu Asn Ser Ile Arg Arg Glu Trp Glu

340 345 350

Val Lys Ala Arg Leu Glu Ile Asp Arg Gln Val Gln Asn Glu Leu Asp

355 360 365

Lys Leu Arg Lys Arg Phe Glu Cys Glu Val Gln Asp Arg Val Ala Gln

370 375 380

Glu Val Glu Lys Gln Arg Arg Asn Ala Asn Tyr Arg Glu Asp Ala Ser

385 390 395 400

Leu Arg Ser Ser Gly His Ser Ser Gln Met Ser Ser Ser Asn Ser Glu

405 410 415

Asp Ser Asp Phe Pro Ser Ser Thr Asp Ile Ser Gln Leu Ser Leu Glu

420 425 430

Ser Pro Thr Asn Lys Ala Ala Lys Leu Pro Lys Lys Glu Ser Arg Thr

435 440 445

Pro Phe Thr Arg Ser Lys Thr Val Val Asp Ser Pro Met Asp Ile Gln

450 455 460

Met Ala Glu Pro Ser Pro Ile Ser Ile Ala Ser Leu Ser Leu Ser Pro

465 470 475 480

Arg Arg Thr Ser Ala Thr Tyr Ser Gly Lys Asn Ile Phe Ala Glu Gly

485 490 495

Glu Arg Lys Arg Pro Lys Phe Glu Pro Thr Leu Ala Tyr Ser Asp Asp

500 505 510

Glu Asp Asp Thr Pro Glu Leu Pro Ser Pro Thr Arg Pro Lys Val Lys

515 520 525

Pro Asp Pro Phe Lys Ala Pro Ser Arg Pro Leu Leu Arg Gln Asn Thr

530 535 540

Thr Ala Leu Met Gln Lys Leu Ser Thr Gln Pro Pro Ile Phe Pro Ala

545 550 555 560

Asn Pro Ser Arg Leu Pro Gln Met Ser Ala Pro Asp Val Arg Glu Ser

565 570 575

Lys Ser Arg Ser Pro His Arg Arg Leu Ser Lys Ile Pro Ser Ser Ala

580 585 590

Asn Leu Ala Ala Asp Ala Gly Ser Pro Thr Arg Lys Asn Gly Val Lys

595 600 605

Ser Ser Pro Ser Lys Met Asn Gly Gly Asp Glu Met Phe Lys Ala Val

610 615 620

Met Gln Arg Asn Met Gly Gly Arg Thr Leu Val Glu Leu Ala Gln Ala

625 630 635 640

Arg Ala Gly Gly Arg Pro Ile Asp Glu Val Lys Arg Cys Ala Ser Asp

645 650 655

Ser Arg Ser Gly Cys Ser Val Pro Met Lys Ser Ala Asp Arg Asp Pro

660 665 670

Pro Ala Val Trp Asp Pro Glu Arg Asp Glu Met Pro Ser Pro Phe Leu

675 680 685

Ala Arg Gly Arg Lys Val Ile Arg Asn Leu Arg

690 695

19

346

Human

19

Met Pro Ser Pro Leu Ala Leu Arg Pro Tyr Leu Arg Ser Glu Phe Ser

1 5 10 15

Pro Ser Val Asp Ala Arg Pro Cys Ser Ser Pro Ser Glu Leu Pro Ala

20 25 30

Lys Leu Leu Leu Gly Ala Thr Leu Pro Arg Ala Pro Arg Leu Pro Arg

35 40 45

Arg Leu Ala Trp Cys Ser Ile Asp Trp Glu Gln Val Cys Leu Leu Gln

50 55 60

Arg Leu Gly Ala Gly Gly Phe Gly Ser Val Tyr Lys Ala Thr Tyr Arg

65 70 75 80

Gly Val Pro Val Ala Ile Lys Gln Val Asn Lys Cys Thr Lys Asn Arg

85 90 95

Leu Ala Ser Arg Arg Ser Phe Trp Ala Glu Leu Asn Val Ala Arg Leu

100 105 110

Arg His Asp Asn Ile Val Arg Val Val Ala Ala Ser Thr Arg Thr Pro

115 120 125

Ala Gly Ser Asn Ser Leu Gly Thr Ile Ile Met Glu Phe Gly Gly Asn

130 135 140

Val Thr Leu His Gln Val Ile Tyr Gly Ala Ala Gly His Pro Glu Gly

145 150 155 160

Asp Ala Gly Glu Pro His Cys Arg Thr Gly Gly Gln Leu Ser Leu Gly

165 170 175

Lys Cys Leu Lys Tyr Ser Leu Asp Val Val Asn Gly Leu Leu Phe Leu

180 185 190

His Ser Gln Ser Ile Val His Leu Asp Leu Lys Pro Ala Asn Ile Leu

195 200 205

Ile Ser Glu Gln Asp Val Cys Lys Ile Ser Asp Phe Gly Cys Ser Glu

210 215 220

Lys Leu Glu Asp Leu Leu Cys Phe Gln Thr Pro Ser Tyr Pro Leu Gly

225 230 235 240

Gly Thr Tyr Thr His Arg Ala Pro Glu Leu Leu Lys Gly Glu Gly Val

245 250 255

Thr Pro Lys Ala Asp Ile Tyr Ser Phe Ala Ile Thr Leu Trp Gln Met

260 265 270

Thr Thr Lys Gln Ala Pro Tyr Ser Gly Glu Arg Gln His Ile Leu Tyr

275 280 285

Ala Val Val Ala Tyr Asp Leu Arg Pro Ser Leu Ser Ala Ala Val Phe

290 295 300

Glu Asp Ser Leu Pro Gly Gln Arg Leu Gly Asp Val Ile Gln Arg Cys

305 310 315 320

Trp Arg Pro Ser Ala Ala Gln Arg Pro Ser Ala Arg Leu Leu Leu Val

325 330 335

Asp Leu Thr Ser Leu Lys Ala Glu Leu Gly

340 345

20

337

Human

20

Met Ala Ser Ser Ser Gly Ser Lys Ala Glu Phe Ile Val Gly Gly Lys

1 5 10 15

Tyr Lys Leu Val Arg Lys Ile Gly Ser Gly Ser Phe Gly Asp Ile Tyr

20 25 30

Leu Ala Ile Asn Ile Thr Asn Gly Glu Glu Val Ala Leu Lys Leu Glu

35 40 45

Ser Gln Lys Ala Arg His Pro Gln Leu Leu Tyr Glu Ser Lys Leu Tyr

50 55 60

Lys Ile Leu Gln Gly Gly Val Gly Ile Pro His Ile Arg Trp Tyr Gly

65 70 75 80

Gln Glu Lys Asp Tyr Asn Val Leu Val Met Asp Leu Leu Gly Pro Ser

85 90 95

Leu Glu Asp Leu Phe Asn Phe Cys Ser Arg Arg Phe Thr Met Lys Thr

100 105 110

Val Leu Met Leu Ala Asp Gln Met Ile Ser Arg Ile Glu Tyr Val His

115 120 125

Thr Lys Asn Phe Ile His Arg Asp Ile Lys Pro Asp Asn Phe Leu Met

130 135 140

Gly Ile Gly Arg His Cys Asn Lys Leu Phe Leu Ile Asp Phe Gly Leu

145 150 155 160

Ala Lys Lys Tyr Arg Asp Asn Arg Thr Arg Gln His Ile Pro Tyr Arg

165 170 175

Glu Asp Lys Asn Leu Thr Gly Thr Ala Arg Tyr Ala Ser Ile Asn Ala

180 185 190

His Leu Gly Ile Glu Gln Ser Arg Arg Asp Asp Met Glu Ser Leu Gly

195 200 205

Tyr Val Leu Met Tyr Phe Asn Arg Thr Ser Leu Pro Trp Gln Gly Leu

210 215 220

Lys Ala Ala Thr Lys Lys Gln Lys Tyr Glu Lys Ile Ser Glu Lys Lys

225 230 235 240

Met Ser Thr Pro Val Glu Val Leu Cys Lys Gly Phe Pro Ala Glu Phe

245 250 255

Ala Met Tyr Leu Asn Tyr Cys Arg Gly Leu Arg Phe Glu Glu Ala Pro

260 265 270

Asp Tyr Met Tyr Leu Arg Gln Leu Phe Arg Ile Leu Phe Arg Thr Leu

275 280 285

Asn His Gln Tyr Asp Tyr Thr Phe Asp Trp Thr Met Leu Lys Gln Lys

290 295 300

Ala Ala Gln Gln Ala Ala Ser Ser Ser Gly Gln Gly Gln Gln Ala Gln

305 310 315 320

Thr Pro Thr Gly Lys Gln Thr Asp Lys Ser Lys Ser Asn Met Lys Gly

325 330 335

Phe

21

336

Bovine

21

Met Ala Ser Ser Ser Arg Pro Lys Thr Asp Val Leu Val Gly Gly Arg

1 5 10 15

Tyr Lys Leu Val Arg Glu Ile Gly Phe Gly Ser Phe Gly His Val Tyr

20 25 30

Leu Ala Ile Asp Leu Thr Asn His Glu Gln Val Ala Val Lys Leu Glu

35 40 45

Ser Glu Asn Thr Arg Gln Pro Arg Leu Leu His Glu Lys Glu Leu Tyr

50 55 60

Asn Phe Leu Gln Gly Gly Val Gly Ile Pro Gln Ile Arg Trp Tyr Gly

65 70 75 80

Gln Glu Thr Asp Tyr Asn Val Leu Val Met Asp Leu Leu Gly Pro Ser

85 90 95

Leu Glu Asp Leu Phe Asn Phe Cys Ser Arg Arg Phe Ser Met Lys Thr

100 105 110

Val Leu Met Leu Ala Asp Gln Met Ile Ser Arg Ile Glu Tyr Val His

115 120 125

Ser Arg Asn Leu Ile His Arg Asp Ile Lys Pro Asp Asn Phe Leu Met

130 135 140

Gly Thr Gly Pro Gln Trp Lys Lys Leu Phe Leu Val Asp Phe Gly Leu

145 150 155 160

Ala Lys Lys Tyr Arg Asp Asn Arg Thr Gly Gln His Ile Pro His Arg

165 170 175

Ser Gly Lys Ser Phe Ile Gly Thr Pro Phe Cys Ala Ser Ile Ser Ala

180 185 190

His Leu Gly Ile Glu Gln Ser Arg Arg Asp Asp Met Glu Ser Ile Gly

195 200 205

Tyr Val Leu Met Tyr Phe Asn Arg Gly Ser Leu Pro Trp Gln Gly Leu

210 215 220

Lys Ala Ala Thr Leu Lys Gln Lys Cys Glu Lys Ile Ser Glu Met Lys

225 230 235 240

Met Thr Thr Pro Val Asp Val Leu Cys Lys Gly Phe Pro Ile Glu Phe

245 250 255

Ala Met Tyr Leu Lys Tyr Cys Leu Arg Leu Ser Phe Glu Glu Ala Pro

260 265 270

Asp Tyr Arg Tyr Leu Arg Gln Leu Phe Arg Leu Leu Phe Arg Lys Leu

275 280 285

Ser Tyr Gln His Asp Tyr Ala Phe Asp Trp Ile Val Leu Lys Gln Lys

290 295 300

Ala Glu Gln Gln Ala Ser Ser Ser Ser Gly Glu Gly Gln Gln Ala Gln

305 310 315 320

Thr Pro Thr Gly Lys Ser Asp Asn Thr Lys Ser Glu Met Lys His Ser

325 330 335

22

415

Human

22

Met Glu Leu Arg Val Gly Asn Arg Tyr Arg Leu Gly Arg Lys Ile Gly

1 5 10 15

Ser Gly Ser Phe Gly Asp Ile Tyr Leu Gly Thr Asp Ile Ala Ala Gly

20 25 30

Glu Glu Val Ala Ile Lys Leu Glu Cys Val Lys Thr Lys His Pro Gln

35 40 45

Leu His Ile Glu Ser Lys Ile Tyr Lys Met Met Gln Gly Gly Val Gly

50 55 60

Ile Pro Thr Ile Arg Trp Cys Gly Ala Glu Gly Asp Tyr Asn Val Met

65 70 75 80

Val Met Glu Leu Leu Gly Pro Ser Leu Glu Asp Leu Phe Asn Phe Cys

85 90 95

Ser Arg Lys Phe Ser Leu Lys Thr Val Leu Leu Leu Ala Asp Gln Met

100 105 110

Ile Ser Arg Ile Glu Tyr Ile His Ser Lys Asn Phe Ile His Arg Asp

115 120 125

Val Lys Pro Asp Asn Phe Leu Met Gly Leu Gly Lys Lys Gly Asn Leu

130 135 140

Val Tyr Ile Ile Asp Phe Gly Leu Ala Lys Lys Tyr Arg Asp Ala Arg

145 150 155 160

Thr His Gln His Ile Pro Tyr Arg Glu Asn Lys Asn Leu Thr Gly Thr

165 170 175

Ala Arg Tyr Ala Ser Ile Asn Thr His Leu Gly Ile Glu Gln Ser Arg

180 185 190

Arg Asp Asp Leu Glu Ser Leu Gly Tyr Val Leu Met Tyr Phe Asn Leu

195 200 205

Gly Ser Leu Pro Trp Gln Gly Leu Lys Ala Ala Thr Lys Arg Gln Lys

210 215 220

Tyr Glu Arg Ile Ser Glu Lys Lys Met Ser Thr Pro Ile Glu Val Leu

225 230 235 240

Cys Lys Gly Tyr Pro Ser Glu Phe Ala Thr Tyr Leu Asn Phe Cys Arg

245 250 255

Ser Leu Arg Phe Asp Asp Lys Pro Asp Tyr Ser Tyr Leu Arg Gln Leu

260 265 270

Phe Arg Asn Leu Phe His Arg Gln Gly Phe Ser Tyr Asp Tyr Val Phe

275 280 285

Asp Trp Asn Met Leu Lys Phe Gly Ala Ser Arg Ala Ala Asp Asp Ala

290 295 300

Glu Arg Glu Arg Arg Asp Arg Glu Glu Arg Leu Arg His Ser Arg Asn

305 310 315 320

Pro Ala Thr Arg Gly Leu Pro Ser Thr Asp Ser Gly Arg Leu Arg Gly

325 330 335

Thr Gln Glu Val Ala Pro Pro Thr Pro Leu Thr Pro Thr Ser His Thr

340 345 350

Ala Asn Thr Ser Pro Arg Pro Val Ser Gly Met Glu Arg Glu Arg Lys

355 360 365

Val Ser Met Arg Leu His Arg Gly Ala Pro Val Asn Ile Ser Ser Ser

370 375 380

Asp Leu Thr Gly Arg Gln Asp Thr Ser Arg Met Ser Thr Ser Gln Ile

385 390 395 400

Pro Gly Arg Val Ala Ser Ser Gly Leu Gln Ser Val Val His Arg

405 410 415

23

538

Yeast

23

Met Ser Met Pro Ile Ala Ser Thr Thr Leu Ala Val Asn Asn Leu Thr

1 5 10 15

Asn Ile Asn Gly Asn Ala Asn Phe Asn Val Gln Ala Asn Lys Gln Leu

20 25 30

His His Gln Ala Val Asp Ser Pro Ala Arg Ser Ser Met Thr Ala Thr

35 40 45

Thr Ala Ala Asn Ser Asn Ser Asn Ser Ser Arg Asp Asp Ser Thr Ile

50 55 60

Val Gly Leu His Tyr Lys Ile Gly Lys Lys Ile Gly Glu Gly Ser Phe

65 70 75 80

Gly Val Leu Phe Glu Gly Thr Asn Met Ile Asn Gly Val Pro Val Ala

85 90 95

Ile Lys Phe Glu Pro Arg Lys Thr Glu Ala Pro Gln Leu Arg Asp Glu

100 105 110

Tyr Lys Thr Tyr Lys Ile Leu Asn Gly Thr Pro Asn Ile Pro Tyr Ala

115 120 125

Tyr Tyr Phe Gly Gln Glu Gly Leu His Asn Ile Leu Val Ile Asp Leu

130 135 140

Leu Gly Pro Ser Leu Glu Asp Leu Phe Asp Trp Cys Gly Arg Lys Phe

145 150 155 160

Ser Val Lys Thr Val Val Gln Val Ala Val Gln Met Ile Thr Leu Ile

165 170 175

Glu Asp Leu His Ala His Asp Leu Ile Tyr Arg Asp Ile Lys Pro Asp

180 185 190

Asn Phe Leu Ile Gly Arg Pro Gly Gln Pro Asp Ala Asn Asn Ile His

195 200 205

Leu Ile Asp Phe Gly Met Ala Lys Gln Tyr Arg Asp Pro Lys Thr Lys

210 215 220

Gln His Ile Pro Tyr Arg Glu Lys Lys Ser Leu Ser Gly Thr Ala Arg

225 230 235 240

Tyr Met Ser Ile Asn Thr His Leu Gly Arg Glu Gln Ser Arg Arg Asp

245 250 255

Asp Met Glu Ala Leu Gly His Val Phe Phe Tyr Phe Leu Arg Gly His

260 265 270

Leu Pro Trp Gln Gly Leu Lys Ala Pro Asn Asn Lys Gln Lys Tyr Glu

275 280 285

Lys Ile Gly Glu Lys Lys Arg Ser Thr Asn Val Tyr Asp Leu Ala Gln

290 295 300

Gly Leu Pro Val Gln Phe Gly Arg Tyr Leu Glu Ile Val Arg Ser Leu

305 310 315 320

Ser Phe Glu Glu Cys Pro Asp Tyr Glu Gly Tyr Arg Lys Leu Leu Leu

325 330 335

Ser Val Leu Asp Asp Leu Gly Glu Thr Ala Asp Gly Gln Tyr Asp Trp

340 345 350

Met Lys Leu Asn Asp Gly Arg Gly Trp Asp Leu Asn Ile Asn Lys Lys

355 360 365

Pro Asn Leu His Gly Tyr Gly His Pro Asn Pro Pro Asn Glu Lys Ser

370 375 380

Arg Lys His Arg Asn Lys Gln Leu Gln Met Gln Gln Leu Gln Met Gln

385 390 395 400

Gln Leu Gln Gln Gln Gln Gln Gln Gln Gln Tyr Ala Gln Lys Thr Glu

405 410 415

Ala Asp Met Arg Asn Ser Gln Tyr Lys Pro Lys Leu Asp Pro Thr Ser

420 425 430

Tyr Glu Ala Tyr Gln His Gln Thr Gln Gln Lys Tyr Leu Gln Glu Gln

435 440 445

Gln Lys Arg Gln Gln Gln Gln Lys Leu Gln Glu Gln Gln Leu Gln Glu

450 455 460

Gln Gln Leu Gln Gln Gln Gln Gln Gln Gln Gln Gln Leu Arg Ala Thr

465 470 475 480

Gly Gln Pro Pro Ser Gln Pro Gln Ala Gln Thr Gln Ser Gln Gln Phe

485 490 495

Gly Ala Arg Tyr Gln Pro Gln Gln Gln Pro Ser Ala Ala Leu Arg Thr

500 505 510

Pro Glu Gln His Pro Asn Asp Asp Asn Ser Ser Leu Ala Ala Ser His

515 520 525

Lys Gly Phe Phe Gln Lys Leu Gly Cys Cys

530 535

24

546

Yeast

24

Met Ser Gln Val Gln Ser Pro Leu Thr Ala Thr Asn Ser Gly Leu Ala

1 5 10 15

Val Asn Asn Asn Thr Met Asn Ser Gln Met Pro Asn Arg Ser Asn Val

20 25 30

Arg Leu Val Asn Gly Thr Leu Pro Pro Ser Leu His Val Ser Ser Asn

35 40 45

Leu Asn His Asn Thr Gly Asn Ser Ser Ala Ser Tyr Ser Gly Ser Gln

50 55 60

Ser Arg Asp Asp Ser Thr Ile Val Gly Leu His Tyr Lys Ile Gly Lys

65 70 75 80

Lys Ile Gly Glu Gly Ser Phe Gly Val Leu Phe Glu Gly Thr Asn Met

85 90 95

Ile Asn Gly Leu Pro Val Ala Ile Lys Phe Glu Pro Arg Lys Thr Glu

100 105 110

Ala Pro Gln Leu Lys Asp Glu Tyr Arg Thr Tyr Lys Ile Leu Ala Gly

115 120 125

Thr Pro Gly Ile Pro Gln Glu Tyr Tyr Phe Gly Gln Glu Gly Leu His

130 135 140

Asn Ile Leu Val Ile Asp Leu Leu Gly Pro Ser Leu Glu Asp Leu Phe

145 150 155 160

Asp Trp Cys Gly Arg Arg Phe Ser Val Lys Thr Val Val Gln Val Ala

165 170 175

Val Gln Met Ile Thr Leu Ile Glu Asp Leu His Ala His Asp Leu Ile

180 185 190

Tyr Arg Asp Ile Lys Pro Asp Asn Phe Leu Ile Gly Arg Pro Gly Gln

195 200 205

Pro Asp Ala Asn Lys Val His Leu Ile Asp Phe Gly Met Ala Lys Gln

210 215 220

Tyr Arg Asp Pro Lys Thr Lys Gln His Ile Pro Tyr Arg Glu Lys Lys

225 230 235 240

Ser Leu Ser Gly Thr Ala Arg Tyr Met Ser Ile Asn Thr His Leu Gly

245 250 255

Arg Glu Gln Ser Arg Arg Asp Asp Met Glu Ala Met Gly His Val Phe

260 265 270

Phe Tyr Phe Leu Arg Gly Gln Leu Pro Trp Gln Gly Leu Lys Ala Pro

275 280 285

Asn Asn Lys Gln Lys Tyr Glu Lys Ile Gly Glu Lys Lys Arg Leu Thr

290 295 300

Asn Val Tyr Asp Leu Ala Gln Gly Leu Pro Ile Gln Phe Gly Arg Tyr

305 310 315 320

Leu Glu Ile Val Arg Asn Leu Ser Phe Glu Glu Thr Pro Asp Tyr Glu

325 330 335

Gly Tyr Arg Met Leu Leu Leu Ser Val Leu Asp Asp Leu Gly Glu Thr

340 345 350

Ala Asp Gly Gln Tyr Asp Trp Met Lys Leu Asn Gly Gly Arg Gly Trp

355 360 365

Asp Leu Ser Ile Asn Lys Lys Pro Asn Leu His Gly Tyr Gly His Pro

370 375 380

Asn Pro Pro Asn Glu Lys Ser Lys Arg His Arg Ser Lys Asn His Gln

385 390 395 400

Tyr Ser Ser Pro Asp His His His His Tyr Asn Gln Gln Gln Gln Gln

405 410 415

Gln Gln Ala Gln Ala Gln Ala Gln Ala Gln Ala Gln Ala Lys Val Gln

420 425 430

Gln Gln Gln Leu Gln Gln Ala Gln Ala Gln Gln Gln Ala Asn Arg Tyr

435 440 445

Gln Leu Gln Pro Asp Asp Ser His Tyr Asp Glu Glu Arg Glu Ala Ser

450 455 460

Lys Leu Asp Pro Thr Ser Tyr Glu Ala Tyr Gln Gln Gln Thr Gln Gln

465 470 475 480

Lys Tyr Ala Gln Gln Gln Gln Lys Gln Met Gln Gln Lys Ser Lys Gln

485 490 495

Phe Ala Asn Thr Gly Ala Asn Gly Gln Thr Asn Lys Tyr Pro Tyr Asn

500 505 510

Ala Gln Pro Thr Ala Asn Asp Glu Gln Asn Ala Lys Asn Ala Ala Gln

515 520 525

Asp Arg Asn Ser Asn Lys Ser Ser Lys Gly Phe Phe Ser Lys Leu Gly

530 535 540

Cys Cys

545

25

494

Yeast

25

Met Asp Leu Arg Val Gly Arg Lys Phe Arg Ile Gly Arg Lys Ile Gly

1 5 10 15

Ser Gly Ser Phe Gly Asp Ile Tyr His Gly Thr Asn Leu Ile Ser Gly

20 25 30

Glu Glu Val Ala Ile Lys Leu Glu Ser Ile Arg Ser Arg His Pro Gln

35 40 45

Leu Asp Tyr Glu Ser Arg Val Tyr Arg Tyr Leu Ser Gly Gly Val Gly

50 55 60

Ile Pro Phe Ile Arg Trp Phe Gly Arg Glu Gly Glu Tyr Asn Ala Met

65 70 75 80

Val Ile Asp Leu Leu Gly Pro Ser Leu Glu Asp Leu Phe Asn Tyr Cys

85 90 95

His Arg Arg Phe Ser Phe Lys Thr Val Ile Met Leu Ala Leu Gln Met

100 105 110

Phe Cys Arg Ile Gln Tyr Ile His Gly Arg Ser Phe Ile His Arg Asp

115 120 125

Ile Lys Pro Asp Asn Phe Leu Met Gly Val Gly Arg Arg Gly Ser Thr

130 135 140

Val His Val Ile Asp Phe Gly Leu Ser Lys Lys Tyr Arg Asp Phe Asn

145 150 155 160

Thr His Arg His Ile Pro Tyr Arg Glu Asn Lys Ser Leu Thr Gly Thr

165 170 175

Ala Arg Tyr Ala Ser Val Asn Thr His Leu Gly Ile Glu Gln Ser Arg

180 185 190

Arg Asp Asp Leu Glu Ser Leu Gly Tyr Val Leu Ile Tyr Phe Cys Lys

195 200 205

Gly Ser Leu Pro Trp Gln Gly Leu Lys Ala Thr Thr Lys Lys Gln Lys

210 215 220

Tyr Asp Arg Ile Met Glu Lys Lys Leu Asn Val Ser Val Glu Thr Leu

225 230 235 240

Cys Ser Gly Leu Pro Leu Glu Phe Gln Glu Tyr Met Ala Tyr Cys Lys

245 250 255

Asn Leu Lys Phe Asp Glu Lys Pro Asp Tyr Leu Phe Leu Ala Arg Leu

260 265 270

Phe Lys Asp Leu Ser Ile Lys Leu Glu Tyr His Asn Asp His Leu Phe

275 280 285

Asp Trp Thr Met Leu Arg Tyr Thr Lys Ala Met Val Glu Lys Gln Arg

290 295 300

Asp Leu Leu Ile Glu Lys Gly Asp Leu Asn Ala Asn Ser Asn Ala Ala

305 310 315 320

Ser Ala Ser Asn Ser Thr Asp Asn Lys Ser Glu Thr Phe Asn Lys Ile

325 330 335

Lys Leu Leu Ala Met Lys Lys Phe Pro Thr His Phe His Tyr Tyr Lys

340 345 350

Asn Glu Asp Lys His Asn Pro Ser Pro Glu Glu Ile Lys Gln Gln Thr

355 360 365

Ile Leu Asn Asn Asn Ala Ala Ser Ser Leu Pro Glu Glu Leu Leu Asn

370 375 380

Ala Leu Asp Lys Gly Met Glu Asn Leu Arg Gln Gln Gln Pro Gln Gln

385 390 395 400

Gln Val Gln Ser Ser Gln Pro Gln Pro Gln Pro Gln Gln Leu Gln Gln

405 410 415

Gln Pro Asn Gly Gln Arg Pro Asn Tyr Tyr Pro Glu Pro Leu Leu Gln

420 425 430

Gln Gln Gln Arg Asp Ser Gln Glu Gln Gln Gln Gln Val Pro Met Ala

435 440 445

Thr Thr Arg Ala Thr Gln Tyr Pro Pro Gln Ile Asn Ser Asn Asn Phe

450 455 460

Asn Thr Asn Gln Ala Ser Val Pro Pro Gln Met Arg Ser Asn Pro Gln

465 470 475 480

Gln Pro Pro Gln Asp Lys Pro Ala Gly Gln Ser Ile Trp Leu

485 490

26

736

Yeast

26

Met Ser Gln Asn Ile Gln Ile Gly Thr Arg Lys Arg Ser Arg Ala Asn

1 5 10 15

Met Asn Asn Ser Thr Thr Thr Gly Pro Ala Asn Asn Thr Ser Ser Asn

20 25 30

Lys Thr Phe Leu Asp Asn Phe Glu Glu Thr Arg Thr Asn Lys Leu Leu

35 40 45

Asp Glu Met Phe Ala Arg Gln Asn Ser Phe Leu Thr Asp Asn Leu Arg

50 55 60

Asn Ser Leu Asp Leu Asn Gln Ala Asp Asn Pro Leu Arg Pro Arg Gln

65 70 75 80

His Gln His Gln Leu Phe Leu Asp Asn Glu Asn Ala Ile Glu Leu Asp

85 90 95

Glu Glu Pro Arg Ile Ile Asn Thr Thr Ile Asn Asn Ser Asn Asn His

100 105 110

Asn Ser Ser Arg Val Asp Glu Asp Ala Asp Asp Asp Ile Ile Phe Ile

115 120 125

Lys Glu Gln Pro Ile Gln Phe Ser Ser Pro Leu Ile Leu Pro Ser Ser

130 135 140

Ser Ser Ile Asn Asn Asn Asn Asn Ile Val Thr Ser Asn Asn Pro Gly

145 150 155 160

Cys Gly Thr Ala Ala Thr Ser Asn Ser Thr Tyr Ile Thr Thr Pro Lys

165 170 175

Lys Phe Lys Lys Gln Arg Thr Ile Ser Leu Pro Gln Leu Pro Leu Ser

180 185 190

Lys Leu Ser Tyr Gln Ser Asn Tyr Phe Asn Val Pro Asp Gln Thr Asn

195 200 205

Ala Ile Val Pro Arg Val Thr Gln Thr Glu Asn Glu Leu Leu His Leu

210 215 220

Thr Gly Ser Cys Ala Lys Thr Leu Glu Gly Asn Lys Ala Val Asn Leu

225 230 235 240

Thr Ile Ala His Ser Thr Ser Pro Phe Ser Asn Pro Pro Ala Gln Ile

245 250 255

Ala Ser Leu Pro Gln Ser Asn Leu Lys Lys Gln Ile Gly Ser Ser Leu

260 265 270

Arg Lys Phe Thr Ser Asn Gly Ser Ser Glu Ser Ala Ser Ser Asn Lys

275 280 285

Ser Asn Phe Lys Thr Asp Lys Asp Gly His Tyr Val Tyr Gln Glu Asn

290 295 300

Asp Ile Phe Gly Ser Gly Gly Arg Phe Val Val Lys Asp Leu Leu Gly

305 310 315 320

Gln Gly Thr Phe Gly Lys Val Leu Lys Cys Ile Asp Asn Lys Tyr Glu

325 330 335

Pro Asn Tyr Val Ala Val Lys Val Ile Arg Ala Val Asp Arg Tyr Arg

340 345 350

Glu Ala Ala Lys Thr Glu Leu Arg Ile Leu Gln Thr Ile Leu Asn Asn

355 360 365

Asp Pro Gln Gly Gln Phe Gln Cys Leu Leu Leu Arg Glu Cys Phe Asp

370 375 380

Tyr Lys Asn His Ile Cys Leu Val Thr Asp Leu Tyr Gly Arg Ser Ile

385 390 395 400

Tyr Asp Phe Met Cys Ser Asn Gly Ile Ala Gly Ser Pro Ala Leu Ile

405 410 415

Ser Gly His Cys Arg Gln Leu Ile Arg Ser Val Cys Phe Leu His Asp

420 425 430

Leu Gly Ile Ile His Thr Asp Leu Lys Pro Glu Asn Ile Leu Ile Cys

435 440 445

Asp Glu Thr His Ile Ala Gln Lys Leu Pro Leu Lys Thr Val Gln Ser

450 455 460

Leu Ser Lys Arg Arg Arg Glu Ala Ser Lys Gly Lys Arg Lys Ile Leu

465 470 475 480

Lys Asn Pro Glu Ile Lys Ile Ile Asp Phe Gly Ser Ala Ile Phe His

485 490 495

Tyr Glu Tyr His Pro Pro Val Ile Ser Thr Arg His Tyr Arg Ala Pro

500 505 510

Glu Ile Val Leu Gly Leu Gly Trp Ser Phe Pro Cys Asp Ile Trp Ser

515 520 525

Ile Ala Cys Val Leu Val Glu Leu Val Ile Gly Glu Ser Leu Tyr Pro

530 535 540

Ile His Glu Asn Leu Glu His Met Ala Met Met Gln Arg Ile Asn Gly

545 550 555 560

Thr Pro Phe Pro Thr Asp Ile Ile Asp Lys Met Phe Tyr Lys Ser Lys

565 570 575

His Lys Leu Gly Asn Ser Pro Ser Asp Leu Asn Ser Thr Val Ile Lys

580 585 590

His Phe Asp Arg Lys Thr Leu Ser Leu Gln Trp Pro Glu Lys Asn Lys

595 600 605

Arg Gly Asp Thr Ile Thr Thr Glu Lys Ser Met Lys Arg Val Leu Gln

610 615 620

Ser Cys Asp Arg Leu Asp Ile Tyr Ile Ser Lys Val Leu Lys Gln Asp

625 630 635 640

Tyr Gly Asp Ser Leu Ser Ile Asn Trp Asn Leu Pro Pro Glu Lys Asn

645 650 655

Trp Ser Leu Ile Asn Ser Lys Leu Ala Trp Lys Arg Gln Thr His Ser

660 665 670

Ser Ser Ser Ser Thr Thr Asp Glu Leu Asp Lys Glu Thr Phe Leu Phe

675 680 685

Trp Tyr Trp Phe Ile Asp Leu Leu Arg Lys Met Phe Glu Phe Asp Pro

690 695 700

Thr Lys Arg Ile Thr Ala Lys Asp Ala Leu Asp His Glu Trp Phe Asn

705 710 715 720

Leu Gly Ile Leu Asp Asp Gly Ile Ala Thr Tyr Asn Asn Thr Gln Gly

725 730 735

27

1584

DICDI - Dictyostelium Discoideum

27

Met Ser Phe Lys Phe Phe Lys Lys Lys Leu Val Pro Ser Asn Glu Tyr

1 5 10 15

Arg Trp Asp Leu Arg Lys Ser Asn Ser Leu Thr Leu Asn Ile Glu Asp

20 25 30

Lys Ser Arg Cys Ser Tyr Arg Leu Pro Thr Ser Gly Ser Lys Gly Ile

35 40 45

Ala Lys Ser Thr Gln Pro Phe Ser Ser Ser Phe Thr Tyr Phe Glu Leu

50 55 60

Phe Ile Thr Asn Gly Asn Gly Asp Lys Ile Cys Phe Gly Leu Thr Thr

65 70 75 80

Asn Asp His Pro Ile Glu Val Tyr Pro Gly Asn Tyr Gln Gly Ser Tyr

85 90 95

Gly Tyr Ser Gly Asp Gly Lys Cys Tyr Phe Gly Thr Asn Glu Gly Arg

100 105 110

Val Tyr Gly Pro Ser Phe Ser Ser Gly Asp Val Val Gly Cys Gly Tyr

115 120 125

Asp Ser Ser Ser Lys Thr Leu Tyr Phe Thr Lys Asn Gly Val Tyr Leu

130 135 140

Gly Val Ala Ala Gln Lys Val Asn Leu Ile Gly Leu Tyr Pro Thr Val

145 150 155 160

Gly Leu Gln Asn Pro Gly Glu Ser Val Val Ile Asn Phe Phe Gly Pro

165 170 175

Phe Ser Tyr Arg Gly Ala Pro Glu Lys Pro Ser Lys Gln Ser Thr Ile

180 185 190

Lys Asp Ser Gly Gly Ser Ser Ile Ile Pro Ser Glu Asp Leu Ile Pro

195 200 205

Lys Glu Glu Phe Glu Val Cys Arg Trp Ser Glu Lys Lys Asn Tyr His

210 215 220

Gly Lys His Val Val Val Arg Asn Arg Thr Ala Phe Leu Pro Leu Asp

225 230 235 240

Ser Pro Lys Asp Thr Ile Gly Gly Val Arg Ala Thr Gln Pro Phe Gly

245 250 255

Glu Gly Phe Cys Tyr Phe Glu Val Ile Ile Asp Gln Leu Asp Lys Gly

260 265 270

Gln Leu Ser Ile Gly Leu Ala Asn Leu Glu Tyr Pro Thr Phe Tyr His

275 280 285

Val Gly Trp Met Pro Arg Ser Tyr Gly Tyr His Asn Asp Asp Gly Arg

290 295 300

Lys Phe Arg Trp Arg Glu Glu Pro Gly Val Asn Glu Gly Glu Ser Tyr

305 310 315 320

Gly Ser Ser Tyr Lys Lys Gly Asp Ile Ile Gly Cys Gly Leu Ser Phe

325 330 335

Thr Ser Arg Glu Ile Phe Phe Thr Lys Asn Gly Met Tyr Leu Gly Thr

340 345 350

Ala Phe Ser Asn Val Tyr Gly Val Phe Tyr Pro Ser Val Ala Phe Asn

355 360 365

Glu Pro Gly Ile Ser Ile Thr Gly Val Phe Gly Pro Pro Phe Lys Phe

370 375 380

Ser Gln Val Thr Leu Met Leu Lys Asn Val Asn Ser Thr Ser Ile Leu

385 390 395 400

Val Pro Asn Gly Asn Asn Asn Asn Asn Ser Asn Asn Asn Asn Asn Asn

405 410 415

Asn Asn Asn Asn Ile Ile Gly Asn Gly Lys Ile Thr Thr Thr Thr Thr

420 425 430

Thr Ser Thr Ser Pro Ser Ser Ile Asn Asn Asn Glu Asp Ile Ser Ser

435 440 445

Asn Asn Asn Asn Asn Asn Asn Asn Asn Asn Asn Asn Asn Asn Asn Asn

450 455 460

Asn Asn Asn Asn Asn Asn Asn Asn Asn Asn Asn Asn Asn Asn Ser Asn

465 470 475 480

Ser Ser Asn Thr Asn Asn Asn Asn Ile Asn Asn Thr Thr Asn Asn Asn

485 490 495

Asn Ser Asn Ser Asn Asn Asn Asn Asn Asn Asn Asn Ser Asn Ser Asn

500 505 510

Ser Asn Ser Asn Asn Asn Asn Ile Asn Asn Asn Asn Asn Asn Asn Asn

515 520 525

Asn Asn Asn Asn Ile Tyr Leu Thr Lys Lys Pro Ser Ile Gly Ser Thr

530 535 540

Asp Glu Ser Ser Thr Gly Ser Leu Gly Gly Asn Asn Ser Ser Gly Asn

545 550 555 560

Asn Asn Ser Ser Ser Gly Ser Ile Gly Asn Asn Ser Ser Ile Ile Lys

565 570 575

Gln Arg Ser Pro Pro His Ser Ile Asn Gly Pro Leu Met Leu Pro Pro

580 585 590

Ser Ser Thr Asn Asn Asn Asn Asn Ile Tyr Ser Ser Tyr Asn Ser Thr

595 600 605

Thr Ala Gly Ser Ser Thr Thr Ile Leu Pro Thr Leu Asn His Pro Ile

610 615 620

Phe Gly Asn Thr Thr Ser Asn Asn Asn Ser Ser Ser Thr Leu Ser Val

625 630 635 640

Gly Gly Asn Asn Asn Leu Leu Gly Arg His Cys Gln Ser Leu Pro Ile

645 650 655

Thr Ala Ser Thr Asn His Thr Leu Ser Ser Ser Leu Gly Val Ser Phe

660 665 670

Ser Ser Pro Ser Ser Ser Pro Lys Thr Ser Pro Arg Lys Ile Val Asn

675 680 685

Ser Ser Glu Asp Leu Gly Phe Val Gln Thr Phe Gln Asp Gln Asp Gly

690 695 700

Gln Pro Pro Ser Ala Trp Arg Arg Cys Gly Lys Ser Ile Lys Thr Lys

705 710 715 720

Asp Asp Ile Thr Leu Thr Ile Ile Lys Lys Lys Thr Ser Val Ala Met

725 730 735

Ala Asp Arg Pro Phe Ser Ser Asn Ser Ser Ser Thr Ile Cys Tyr Phe

740 745 750

Glu Val Tyr Leu Glu Gly His Asp Lys Lys Gly Ser Ile Thr Val Gly

755 760 765

Leu Ser His Ser Thr Tyr Pro Phe Ile Lys His Ile Gly Arg Glu Pro

770 775 780

Lys Ser Tyr Gly Phe Ser Ser Glu Gly Glu Lys Tyr Gly Gly Ser Glu

785 790 795 800

Ile Gly Glu Pro Tyr Gly Pro Phe Phe Phe Phe Asp Gly Asp Ser Ile

805 810 815

Ala Ser Ser Cys Val Ile Gly Cys Gly Ile Asn Thr Ser Thr Arg Asp

820 825 830

Ile Phe Phe Thr Lys Asn Gly His Tyr Leu Gly Val Ala Phe Ser Arg

835 840 845

Val Thr Ser Asp Pro Leu Tyr Pro Ser Ile Ser Phe Arg Gly Val Val

850 855 860

Gly Gly Leu Cys Val Ala Thr Phe Pro Gly Gly His Phe Arg Phe Asn

865 870 875 880

Ile Glu Asp Leu Pro Gly Ile Ser Pro Ser Val Trp Thr Glu Ala Leu

885 890 895

Gly Pro Asp Arg Gln Gly Ser Gly Phe Lys Asn Trp Ala Pro Asn Asp

900 905 910

Val Ala Ile Trp Leu Glu Ser Phe Asn Tyr Gly Gln Tyr Arg Lys Asn

915 920 925

Phe Arg Asp Asn Asn Ile Ser Gly Arg His Leu Glu Gly Ile Thr His

930 935 940

Ala Met Leu Lys Asn Asp Leu Gly Ile Glu Pro Tyr Gly His Arg Glu

945 950 955 960

Asp Ile Ile Asn Arg Leu Asn Arg Met Ile Gln Ile Trp Asn Asp Lys

965 970 975

Ser Pro Asp Ser Tyr Pro Lys Ile Ala Ile Asp Ser Ser Asp Lys Ile

980 985 990

Arg Trp Pro Ala Ser Gly Gly Ser Ser Gly Gly Ile Asn Ile Ser Gly

995 1000 1005

Gly Val Val Ile Gly Ser Ser Ser Gly Ser Asp Asp Gly Ile Thr

1010 1015 1020

Glu Ile Ser Ser Ser Ser Lys Asn Ile Arg Pro Tyr Lys Ser Tyr

1025 1030 1035

Thr Gln Lys Glu Ile Glu Asp Arg Asn Arg Arg Ser Thr Ile Ser

1040 1045 1050

Gly Gly Glu Lys Lys Asn Lys Tyr Tyr Ile Asp Asn Gln Met Asp

1055 1060 1065

Pro His Gln Ile Gly Ser Met Asp Ser Asp Gly Leu Leu Pro Asp

1070 1075 1080

Phe Gly Gln Gly Pro Pro Asp Glu Lys Asn Ser Ser Lys Thr Leu

1085 1090 1095

Ser Asn Glu Gln Ile Arg Tyr Leu Gln Gln Arg Lys Asp Glu Pro

1100 1105 1110

Pro Ile Ala Ile Ser Ser Thr Gly Asn Gly Gly Ser Val Ser Ser

1115 1120 1125

Thr Gly Gly Ser Ser Gly Phe Leu Thr Phe Pro Ser Ser Asn Ser

1130 1135 1140

Leu Thr His Pro Pro Gln Arg Asp Lys Pro Thr Gln Glu Phe Thr

1145 1150 1155

His Leu Pro Pro Ile Thr Ser Asn Tyr Lys Gly Ile Thr Asn Thr

1160 1165 1170

Gly Gln Pro His Lys Ser Phe Asp Gln Pro Leu Glu Leu Phe Pro

1175 1180 1185

Arg His Ser Ala Phe Ser Asn Asn Gly Asn Asn Gly Asn Asn Asn

1190 1195 1200

Asn Asn Asn Asn Asn Asn Asn Ile Lys Ala Asn Gln Gln Gln Gln

1205 1210 1215

Gln Gln Ser Ser Tyr Gln Gln Ser Gln Thr Gln Gln Gln Gln Gln

1220 1225 1230

His Ile Thr Ser Thr Ser Thr Ser Thr Thr Asn Lys Trp Ile Asp

1235 1240 1245

Pro Phe Gly Gly Trp Glu Thr Gln Ser Ser Leu Ser His Pro Pro

1250 1255 1260

Ser Arg Pro Pro Pro Pro Pro Pro Pro Pro Pro Gln Leu Pro Val

1265 1270 1275

Arg Ser Glu Tyr Glu Ile Asp Phe Asn Glu Leu Glu Phe Gly Gln

1280 1285 1290

Thr Ile Gly Lys Gly Phe Phe Gly Glu Val Lys Arg Gly Tyr Trp

1295 1300 1305

Arg Glu Thr Asp Val Ala Ile Lys Ile Ile Tyr Arg Asp Gln Phe

1310 1315 1320

Lys Thr Lys Ser Ser Leu Val Met Phe Gln Asn Glu Val Gly Ile

1325 1330 1335

Leu Ser Lys Leu Arg His Pro Asn Val Val Gln Phe Leu Gly Ala

1340 1345 1350

Cys Thr Ala Gly Gly Glu Asp His His Cys Ile Val Thr Glu Trp

1355 1360 1365

Met Gly Gly Gly Ser Leu Arg Gln Phe Leu Thr Asp His Phe Asn

1370 1375 1380

Leu Leu Glu Gln Asn Pro His Ile Arg Leu Lys Leu Ala Leu Asp

1385 1390 1395

Ile Ala Lys Gly Met Asn Tyr Leu His Gly Trp Thr Pro Pro Ile

1400 1405 1410

Leu His Arg Asp Leu Ser Ser Arg Asn Ile Leu Leu Asp His Asn

1415 1420 1425

Ile Asp Pro Lys Asn Pro Val Val Ser Ser Arg Gln Asp Ile Lys

1430 1435 1440

Cys Lys Ile Ser Asp Phe Gly Leu Ser Arg Leu Lys Lys Glu Gln

1445 1450 1455

Ala Ser Gln Met Thr Gln Ser Val Gly Cys Ile Pro Tyr Met Ala

1460 1465 1470

Pro Glu Val Phe Lys Gly Asp Ser Asn Ser Glu Lys Ser Asp Val

1475 1480 1485

Tyr Ser Tyr Gly Met Val Leu Phe Glu Leu Leu Thr Ser Asp Glu

1490 1495 1500

Pro Gln Gln Asp Met Lys Pro Met Lys Met Ala His Leu Ala Ala

1505 1510 1515

Tyr Glu Ser Tyr Arg Pro Pro Ile Pro Leu Thr Thr Ser Ser Lys

1520 1525 1530

Trp Lys Glu Ile Leu Thr Gln Cys Trp Asp Ser Asn Pro Asp Ser

1535 1540 1545

Arg Pro Thr Phe Lys Gln Ile Ile Val His Leu Lys Glu Met Glu

1550 1555 1560

Asp Gln Gly Val Ser Ser Phe Ala Ser Val Pro Val Gln Thr Ile

1565 1570 1575

Asp Thr Gly Val Tyr Ala

1580

28

446

SCHPO - Fission Yeast

28

Met Ser Gly Gln Asn Asn Val Val Gly Val His Tyr Lys Val Gly Arg

1 5 10 15

Arg Ile Gly Glu Gly Ser Phe Gly Val Ile Phe Glu Gly Thr Asn Leu

20 25 30

Leu Asn Asn Gln Gln Val Ala Ile Lys Phe Glu Pro Arg Arg Ser Asp

35 40 45

Ala Pro Gln Leu Arg Asp Glu Tyr Arg Thr Tyr Lys Leu Leu Ala Gly

50 55 60

Cys Thr Gly Ile Pro Asn Val Tyr Tyr Phe Gly Gln Glu Gly Leu His

65 70 75 80

Asn Ile Leu Val Ile Asp Leu Leu Gly Pro Ser Leu Glu Asp Leu Leu

85 90 95

Asp Leu Cys Gly Arg Lys Phe Ser Val Lys Thr Val Ala Met Ala Ala

100 105 110

Lys Gln Met Leu Ala Arg Val Gln Ser Ile His Glu Lys Ser Leu Val

115 120 125

Tyr Arg Asp Ile Lys Pro Asp Asn Phe Leu Ile Gly Arg Pro Asn Ser

130 135 140

Lys Asn Ala Asn Met Ile Tyr Val Val Asp Phe Gly Met Val Lys Phe

145 150 155 160

Tyr Arg Asp Pro Val Thr Lys Gln His Ile Pro Tyr Arg Glu Lys Lys

165 170 175

Asn Leu Ser Gly Thr Ala Arg Tyr Met Ser Ile Asn Thr His Leu Gly

180 185 190

Arg Glu Gln Ser Arg Arg Asp Asp Leu Glu Ala Leu Gly His Val Phe

195 200 205

Met Tyr Phe Leu Arg Gly Ser Leu Pro Trp Gln Gly Leu Lys Ala Ala

210 215 220

Thr Asn Lys Gln Lys Tyr Glu Arg Ile Gly Glu Lys Lys Gln Ser Thr

225 230 235 240

Pro Leu Arg Glu Leu Cys Ala Gly Phe Pro Glu Glu Phe Tyr Lys Tyr

245 250 255

Met His Tyr Ala Arg Asn Leu Ala Phe Asp Ala Thr Pro Asp Tyr Asp

260 265 270

Tyr Leu Gln Gly Leu Phe Ser Lys Val Leu Glu Arg Leu Asn Thr Thr

275 280 285

Glu Asp Glu Asn Phe Asp Trp Asn Leu Leu Asn Asn Gly Lys Gly Trp

290 295 300

Gln Ser Leu Lys Ser Arg Asn Ala Glu Thr Glu Asn Gln Arg Ser Ser

305 310 315 320

Lys Pro Pro Ala Pro Lys Leu Glu Ser Lys Ser Pro Ala Leu Gln Asn

325 330 335

His Ala Ser Thr Gln Asn Val Val Ser Lys Arg Ser Asp Tyr Glu Lys

340 345 350

Pro Phe Ala Glu Pro His Leu Asn Ser Ala Ser Asp Ser Ala Glu Pro

355 360 365

Asn Gln Asn Ser Leu Pro Asn Pro Pro Thr Glu Thr Lys Ala Thr Thr

370 375 380

Thr Val Pro Asp Arg Ser Gly Leu Ala Thr Asn Gln Pro Ala Pro Val

385 390 395 400

Asp Val His Asp Ser Ser Glu Glu Arg Val Thr Arg Glu Gln Val Gln

405 410 415

Asn Ala Thr Lys Glu Thr Glu Ala Pro Lys Lys Lys Lys Ser Phe Trp

420 425 430

Ala Ser Ile Leu Ser Cys Cys Ser Gly Ser Asn Glu Asp Thr

435 440 445

29

298

Human

29

Met Glu Asn Phe Gln Lys Val Glu Lys Ile Gly Glu Gly Thr Tyr Gly

1 5 10 15

Val Val Tyr Lys Ala Arg Asn Lys Leu Thr Gly Glu Val Val Ala Leu

20 25 30

Lys Lys Ile Arg Leu Asp Thr Glu Thr Glu Gly Val Pro Ser Thr Ala

35 40 45

Ile Arg Glu Ile Ser Leu Leu Lys Glu Leu Asn His Pro Asn Ile Val

50 55 60

Lys Leu Leu Asp Val Ile His Thr Glu Asn Lys Leu Tyr Leu Val Phe

65 70 75 80

Glu Phe Leu His Gln Asp Leu Lys Lys Phe Met Asp Ala Ser Ala Leu

85 90 95

Thr Gly Ile Pro Leu Pro Leu Ile Lys Ser Tyr Leu Phe Gln Leu Leu

100 105 110

Gln Gly Leu Ala Phe Cys His Ser His Arg Val Leu His Arg Asp Leu

115 120 125

Lys Pro Gln Asn Leu Leu Ile Asn Thr Glu Gly Ala Ile Lys Leu Ala

130 135 140

Asp Phe Gly Leu Ala Arg Ala Phe Gly Val Pro Val Arg Thr Tyr Thr

145 150 155 160

His Glu Val Val Thr Leu Trp Tyr Arg Ala Pro Glu Ile Leu Leu Gly

165 170 175

Cys Lys Tyr Tyr Ser Thr Ala Val Asp Ile Trp Ser Leu Gly Cys Ile

180 185 190

Phe Ala Glu Met Val Thr Arg Arg Ala Leu Phe Pro Gly Asp Ser Glu

195 200 205

Ile Asp Gln Leu Phe Arg Ile Phe Arg Thr Leu Gly Thr Pro Asp Glu

210 215 220

Val Val Trp Pro Gly Val Thr Ser Met Pro Asp Tyr Lys Pro Ser Phe

225 230 235 240

Pro Lys Trp Ala Arg Gln Asp Phe Ser Lys Val Val Pro Pro Leu Asp

245 250 255

Glu Asp Gly Arg Ser Leu Leu Ser Gln Met Leu His Tyr Asp Pro Asn

260 265 270

Lys Arg Ile Ser Ala Lys Ala Ala Leu Ala His Pro Phe Phe Gln Asp

275 280 285

Val Thr Lys Pro Val Pro His Leu Arg Leu

290 295

30

387

Human

30

Met Thr Arg Asp Glu Ala Leu Pro Asp Ser His Ser Ala Gln Asp Phe

1 5 10 15

Tyr Glu Asn Tyr Glu Pro Lys Glu Ile Leu Gly Arg Gly Val Ser Ser

20 25 30

Val Val Arg Arg Cys Ile His Lys Pro Thr Ser Gln Glu Tyr Ala Val

35 40 45

Lys Val Ile Asp Val Thr Gly Gly Gly Ser Phe Ser Pro Glu Glu Val

50 55 60

Arg Glu Leu Arg Glu Ala Thr Leu Lys Glu Val Asp Ile Leu Arg Lys

65 70 75 80

Val Ser Gly His Pro Asn Ile Ile Gln Leu Lys Asp Thr Tyr Glu Thr

85 90 95

Asn Thr Phe Phe Phe Leu Val Phe Asp Leu Met Lys Arg Gly Glu Leu

100 105 110

Phe Asp Tyr Leu Thr Glu Lys Val Thr Leu Ser Glu Lys Glu Thr Arg

115 120 125

Lys Ile Met Arg Ala Leu Leu Glu Val Ile Cys Thr Leu His Lys Leu

130 135 140

Asn Ile Val His Arg Asp Leu Lys Pro Glu Asn Ile Leu Leu Asp Asp

145 150 155 160

Asn Met Asn Ile Lys Leu Thr Asp Phe Gly Phe Ser Cys Gln Leu Glu

165 170 175

Pro Gly Glu Arg Leu Arg Glu Val Cys Gly Thr Pro Ser Tyr Leu Ala

180 185 190

Pro Glu Ile Ile Glu Cys Ser Met Asn Glu Asp His Pro Gly Tyr Gly

195 200 205

Lys Glu Val Asp Met Trp Ser Thr Gly Val Ile Met Tyr Thr Leu Leu

210 215 220

Ala Gly Ser Pro Pro Phe Trp His Arg Lys Gln Met Leu Met Leu Arg

225 230 235 240

Met Ile Met Ser Gly Asn Tyr Gln Phe Gly Ser Pro Glu Trp Asp Asp

245 250 255

Tyr Ser Asp Thr Val Lys Asp Leu Val Ser Arg Phe Leu Val Val Gln

260 265 270

Pro Gln Asn Arg Tyr Thr Ala Glu Glu Ala Leu Ala His Pro Phe Phe

275 280 285

Gln Gln Tyr Leu Val Glu Glu Val Arg His Phe Ser Pro Arg Gly Lys

290 295 300

Phe Lys Val Ile Ala Leu Thr Val Leu Ala Ser Val Arg Ile Tyr Tyr

305 310 315 320

Gln Tyr Arg Arg Val Lys Pro Val Thr Arg Glu Ile Val Ile Arg Asp

325 330 335

Pro Tyr Ala Leu Arg Pro Leu Arg Arg Leu Ile Asp Ala Tyr Ala Phe

340 345 350

Arg Ile Tyr Gly His Trp Val Lys Lys Gly Gln Gln Gln Asn Arg Ala

355 360 365

Ala Leu Phe Glu Asn Thr Pro Lys Ala Val Leu Leu Ser Leu Ala Glu

370 375 380

Glu Asp Tyr

385

31

370

Human

31

Met Leu Gly Ala Val Glu Gly Pro Arg Trp Lys Gln Ala Glu Asp Ile

1 5 10 15

Arg Asp Ile Tyr Asp Phe Arg Asp Val Leu Gly Thr Gly Ala Phe Ser

20 25 30

Glu Val Ile Leu Ala Glu Asp Lys Arg Thr Gln Lys Leu Val Ala Ile

35 40 45

Lys Cys Ile Ala Lys Glu Ala Leu Glu Gly Lys Glu Gly Ser Met Glu

50 55 60

Asn Glu Ile Ala Val Leu His Lys Ile Lys His Pro Asn Ile Val Ala

65 70 75 80

Leu Asp Asp Ile Tyr Glu Ser Gly Gly His Leu Tyr Leu Ile Met Gln

85 90 95

Leu Val Ser Gly Gly Glu Leu Phe Asp Arg Ile Val Glu Lys Gly Phe

100 105 110

Tyr Thr Glu Arg Asp Ala Ser Arg Leu Ile Phe Gln Val Leu Asp Ala

115 120 125

Val Lys Tyr Leu His Asp Leu Gly Ile Val His Arg Asp Leu Lys Pro

130 135 140

Glu Asn Leu Leu Tyr Tyr Ser Leu Asp Glu Asp Ser Lys Ile Met Ile

145 150 155 160

Ser Asp Phe Gly Leu Ser Lys Met Glu Asp Pro Gly Ser Val Leu Ser

165 170 175

Thr Ala Cys Gly Thr Pro Gly Tyr Val Ala Pro Glu Val Leu Ala Gln

180 185 190

Lys Pro Tyr Ser Lys Ala Val Asp Cys Trp Ser Ile Gly Val Ile Ala

195 200 205

Tyr Ile Leu Leu Cys Gly Tyr Pro Pro Phe Tyr Asp Glu Asn Asp Ala

210 215 220

Lys Leu Phe Glu Gln Ile Leu Lys Ala Glu Tyr Glu Phe Asp Ser Pro

225 230 235 240

Tyr Trp Asp Asp Ile Ser Asp Ser Ala Lys Asp Phe Ile Arg His Leu

245 250 255

Met Glu Lys Asp Pro Glu Lys Arg Phe Thr Cys Glu Gln Ala Leu Gln

260 265 270

His Pro Trp Ile Ala Gly Asp Thr Ala Leu Asp Lys Asn Ile His Gln

275 280 285

Ser Val Ser Glu Gln Ile Lys Lys Asn Phe Ala Lys Ser Lys Trp Lys

290 295 300

Gln Ala Phe Asn Ala Thr Ala Val Val Arg His Met Arg Lys Leu Gln

305 310 315 320

Leu Gly Thr Ser Gln Glu Gly Gln Gly Gln Thr Ala Ser His Gly Glu

325 330 335

Leu Leu Thr Pro Val Ala Gly Gly Pro Ala Ala Gly Cys Cys Cys Arg

340 345 350

Asp Cys Cys Val Glu Pro Gly Thr Glu Leu Ser Pro Thr Leu Pro His

355 360 365

Gln Leu

370

32

372

Human

32

Met Ser Gly Pro Arg Ala Gly Phe Tyr Arg Gln Glu Leu Asn Lys Thr

1 5 10 15

Val Trp Glu Val Pro Gln Arg Leu Gln Gly Leu Arg Pro Val Gly Ser

20 25 30

Gly Ala Tyr Gly Ser Val Cys Ser Ala Tyr Asp Ala Arg Leu Arg Gln

35 40 45

Lys Val Ala Val Lys Lys Leu Ser Arg Pro Phe Gln Ser Leu Ile His

50 55 60

Ala Arg Arg Thr Tyr Arg Glu Leu Arg Leu Leu Lys His Leu Lys His

65 70 75 80

Glu Asn Val Ile Gly Leu Leu Asp Val Phe Thr Pro Ala Thr Ser Ile

85 90 95

Glu Asp Phe Ser Glu Val Tyr Leu Val Thr Thr Leu Met Gly Ala Asp

100 105 110

Leu Asn Asn Ile Val Lys Cys Gln Ala Gly Ala His Gln Gly Ala Arg

115 120 125

Leu Ala Leu Asp Glu His Val Gln Phe Leu Val Tyr Gln Leu Leu Arg

130 135 140

Gly Leu Lys Tyr Ile His Ser Ala Gly Ile Ile His Arg Asp Leu Lys

145 150 155 160

Pro Ser Asn Val Ala Val Asn Glu Asp Cys Glu Leu Arg Ile Leu Asp

165 170 175

Phe Gly Leu Ala Arg Gln Ala Asp Glu Glu Met Thr Gly Tyr Val Ala

180 185 190

Thr Arg Trp Tyr Arg Ala Pro Glu Ile Met Leu Asn Trp Met His Tyr

195 200 205

Asn Gln Thr Val Asp Ile Trp Ser Val Gly Cys Ile Met Ala Glu Leu

210 215 220

Leu Gln Gly Lys Ala Leu Phe Pro Gly Ser Asp Tyr Ile Asp Gln Leu

225 230 235 240

Lys Arg Ile Met Glu Val Val Gly Thr Pro Ser Pro Glu Val Leu Ala

245 250 255

Lys Ile Ser Ser Glu His Ala Arg Thr Tyr Ile Gln Ser Leu Pro Pro

260 265 270

Met Pro Gln Lys Asp Leu Ser Ser Ile Phe Arg Gly Ala Asn Pro Leu

275 280 285

Ala Ile Asp Leu Leu Gly Arg Met Leu Val Leu Asp Ser Asp Gln Arg

290 295 300

Val Ser Ala Ala Glu Ala Leu Ala His Ala Tyr Phe Ser Gln Tyr His

305 310 315 320

Asp Pro Glu Asp Glu Pro Glu Ala Glu Pro Tyr Asp Glu Ser Val Glu

325 330 335

Ala Lys Glu Arg Thr Leu Glu Glu Trp Lys Glu Leu Thr Tyr Gln Glu

340 345 350

Val Leu Ser Phe Lys Pro Pro Glu Pro Pro Lys Pro Pro Gly Ser Leu

355 360 365

Glu Ile Glu Gln

370

33

367

Human

33

Met Ser Ser Pro Pro Pro Ala Arg Ser Gly Phe Tyr Arg Gln Glu Val

1 5 10 15

Thr Lys Thr Ala Trp Glu Val Arg Ala Val Tyr Arg Asp Leu Gln Pro

20 25 30

Val Gly Ser Gly Ala Tyr Gly Ala Val Cys Ser Ala Val Asp Gly Arg

35 40 45

Thr Gly Ala Lys Val Ala Ile Lys Lys Leu Tyr Arg Pro Phe Gln Ser

50 55 60

Glu Leu Phe Ala Lys Arg Ala Tyr Arg Glu Leu Arg Leu Leu Lys His

65 70 75 80

Met Arg His Glu Asn Val Ile Gly Leu Leu Asp Val Phe Thr Pro Asp

85 90 95

Glu Thr Leu Asp Asp Phe Thr Asp Phe Tyr Leu Val Met Pro Phe Met

100 105 110

Gly Thr Asp Leu Gly Lys Leu Met Lys His Glu Lys Leu Gly Glu Asp

115 120 125

Arg Ile Gln Phe Leu Val Tyr Gln Met Leu Lys Gly Leu Arg Tyr Ile

130 135 140

His Ala Ala Gly Ile Ile His Arg Asp Leu Lys Pro Gly Asn Leu Ala

145 150 155 160

Val Asn Glu Asp Cys Glu Leu Lys Ile Leu Asp Phe Gly Leu Ala Arg

165 170 175

Gln Ala Asp Ser Glu Met Thr Gly Tyr Val Val Thr Arg Trp Tyr Arg

180 185 190

Ala Pro Glu Val Ile Leu Asn Trp Met Arg Tyr Thr Gln Thr Val Asp

195 200 205

Ile Trp Ser Val Gly Cys Ile Met Ala Glu Met Ile Thr Gly Lys Thr

210 215 220

Leu Phe Lys Gly Ser Asp His Leu Asp Gln Leu Lys Glu Ile Met Lys

225 230 235 240

Val Thr Gly Thr Pro Pro Ala Glu Phe Val Gln Arg Leu Gln Ser Asp

245 250 255

Glu Ala Lys Asn Tyr Met Lys Gly Leu Pro Glu Leu Glu Lys Lys Asp

260 265 270

Phe Ala Ser Ile Leu Thr Asn Ala Ser Pro Leu Ala Val Asn Leu Leu

275 280 285

Glu Lys Met Leu Val Leu Asp Ala Glu Gln Arg Val Thr Ala Gly Glu

290 295 300

Ala Leu Ala His Pro Tyr Phe Glu Ser Leu His Asp Thr Glu Asp Glu

305 310 315 320

Pro Gln Val Gln Lys Tyr Asp Asp Ser Phe Asp Asp Val Asp Arg Thr

325 330 335

Leu Asp Glu Trp Lys Arg Val Thr Tyr Lys Glu Val Leu Ser Phe Lys

340 345 350

Pro Pro Arg Gln Leu Gly Ala Arg Val Ser Lys Glu Thr Pro Leu

355 360 365

34

364

Human

34

His His His His His His Met Ala Ala Ala Ala Ala Ala Gly Pro Glu

1 5 10 15

Met Val Arg Gly Gln Val Phe Asp Val Gly Pro Arg Tyr Thr Asn Leu

20 25 30

Ser Tyr Ile Gly Glu Gly Ala Tyr Gly Met Val Cys Ser Ala Tyr Asp

35 40 45

Asn Leu Asn Lys Val Arg Val Ala Ile Lys Lys Ile Ser Pro Phe Glu

50 55 60

His Gln Thr Tyr Cys Gln Arg Thr Leu Arg Glu Ile Lys Ile Leu Leu

65 70 75 80

Arg Phe Arg His Glu Asn Ile Ile Gly Ile Asn Asp Ile Ile Arg Ala

85 90 95

Pro Thr Ile Glu Gln Met Lys Asp Val Tyr Ile Val Gln Asp Leu Met

100 105 110

Glu Thr Asp Leu Tyr Lys Leu Leu Lys Thr Gln His Leu Ser Asn Asp

115 120 125

His Ile Cys Tyr Phe Leu Tyr Gln Ile Leu Arg Gly Leu Lys Tyr Ile

130 135 140

His Ser Ala Asn Val Leu His Arg Asp Leu Lys Pro Ser Asn Leu Leu

145 150 155 160

Leu Asn Thr Thr Cys Asp Leu Lys Ile Cys Asp Phe Gly Leu Ala Arg

165 170 175

Val Ala Asp Pro Asp His Asp His Thr Gly Phe Leu Thr Glu Tyr Val

180 185 190

Ala Thr Arg Trp Tyr Arg Ala Pro Glu Ile Met Leu Asn Ser Lys Gly

195 200 205

Tyr Thr Lys Ser Ile Asp Ile Trp Ser Val Gly Cys Ile Leu Ala Glu

210 215 220

Met Leu Ser Asn Arg Pro Ile Phe Pro Gly Lys His Tyr Leu Asp Gln

225 230 235 240

Leu Asn His Ile Leu Gly Ile Leu Gly Ser Pro Ser Gln Glu Asp Leu

245 250 255

Asn Cys Ile Ile Asn Leu Lys Ala Arg Asn Tyr Leu Leu Ser Leu Pro

260 265 270

His Lys Asn Lys Val Pro Trp Asn Arg Leu Phe Pro Asn Ala Asp Ser

275 280 285

Lys Ala Leu Asp Leu Leu Asp Lys Met Leu Thr Phe Asn Pro His Lys

290 295 300

Arg Ile Glu Val Glu Gln Ala Leu Ala His Pro Tyr Leu Glu Gln Tyr

305 310 315 320

Tyr Asp Pro Ser Asp Glu Pro Ile Ala Glu Ala Pro Phe Lys Phe Asp

325 330 335

Met Glu Leu Asp Asp Leu Pro Lys Glu Lys Leu Lys Glu Leu Ile Phe

340 345 350

Glu Glu Thr Ala Arg Phe Gln Pro Gly Tyr Arg Ser

355 360

35

Ala His His His His His His Met Ala Ala Ala Ala Ala Ala Gly Pro

1 5 10 15

Glu Met Val Arg Gly Gln Val Phe Asp Val Gly Pro Arg Tyr Thr Asn

20 25 30

Leu Ser Tyr Ile Gly Glu Gly Ala Tyr Gly Met Val Cys Ser Ala Tyr

35 40 45

Asp Asn Leu Asn Lys Val Arg Val Ala Ile Lys Lys Ile Ser Pro Phe

50 55 60

Glu His Gln Thr Tyr Cys Gln Arg Thr Leu Arg Glu Ile Lys Ile Leu

65 70 75 80

Leu Arg Phe Arg His Glu Asn Ile Ile Gly Ile Asn Asp Ile Ile Arg

85 90 95

Ala Pro Thr Ile Glu Gln Met Lys Asp Val Tyr Ile Val Gln Asp Leu

100 105 110

Met Glu Thr Asp Leu Tyr Lys Leu Leu Lys Thr Gln His Leu Ser Asn

115 120 125

Asp His Ile Cys Tyr Phe Leu Tyr Gln Ile Leu Arg Gly Leu Lys Tyr

130 135 140

Ile His Ser Ala Asn Val Leu His Arg Asp Leu Lys Pro Ser Asn Leu

145 150 155 160

Leu Leu Asn Thr Thr Cys Asp Leu Lys Ile Cys Asp Phe Gly Leu Ala

165 170 175

Arg Val Ala Asp Pro Asp His Asp His Thr Gly Phe Leu Xaa Glu Xaa

180 185 190

Val Ala Thr Arg Trp Tyr Arg Ala Pro Glu Ile Met Leu Asn Ser Lys

195 200 205

Gly Tyr Thr Lys Ser Ile Asp Ile Trp Ser Val Gly Cys Ile Leu Ala

210 215 220

Glu Met Leu Ser Asn Arg Pro Ile Phe Pro Gly Lys His Tyr Leu Asp

225 230 235 240

Gln Leu Asn His Ile Leu Gly Ile Leu Gly Ser Pro Ser Gln Glu Asp

245 250 255

Leu Asn Cys Ile Ile Asn Leu Lys Ala Arg Asn Tyr Leu Leu Ser Leu

260 265 270

Pro His Lys Asn Lys Val Pro Trp Asn Arg Leu Phe Pro Asn Ala Asp

275 280 285

Ser Lys Ala Leu Asp Leu Leu Asp Lys Met Leu Thr Phe Asn Pro His

290 295 300

Lys Arg Ile Glu Val Glu Gln Ala Leu Ala His Pro Tyr Leu Glu Gln

305 310 315 320

Tyr Tyr Asp Pro Ser Asp Glu Pro Ile Ala Glu Ala Pro Phe Lys Phe

325 330 335

Asp Met Glu Leu Asp Asp Leu Pro Lys Glu Lys Leu Lys Glu Leu Ile

340 345 350

Phe Glu Glu Thr Ala Arg Phe Gln Pro Gly Tyr Arg Ser

355 360 365

36

379

Human

36

Gly Ser Ser His His His His His His Ser Ser Gly Leu Val Pro Arg

1 5 10 15

Gly Ser His Met Ser Gln Glu Arg Pro Thr Phe Tyr Arg Gln Glu Leu

20 25 30

Asn Lys Thr Ile Trp Glu Val Pro Glu Arg Tyr Gln Asn Leu Ser Pro

35 40 45

Val Gly Ser Gly Ala Tyr Gly Ser Val Cys Ala Ala Phe Asp Thr Lys

50 55 60

Thr Gly His Arg Val Ala Val Lys Lys Leu Ser Arg Pro Phe Gln Ser

65 70 75 80

Ile Ile His Ala Lys Arg Thr Tyr Arg Glu Leu Arg Leu Leu Lys His

85 90 95

Met Lys His Glu Asn Val Ile Gly Leu Leu Asp Val Phe Thr Pro Ala

100 105 110

Arg Ser Leu Glu Glu Phe Asn Asp Val Tyr Leu Val Thr His Leu Met

115 120 125

Gly Ala Asp Leu Asn Asn Ile Val Lys Cys Gln Lys Leu Thr Asp Asp

130 135 140

His Val Gln Phe Leu Ile Tyr Gln Ile Leu Arg Gly Leu Lys Tyr Ile

145 150 155 160

His Ser Ala Asp Ile Ile His Arg Asp Leu Lys Pro Ser Asn Leu Ala

165 170 175

Val Asn Glu Asp Cys Glu Leu Lys Ile Leu Asp Phe Gly Leu Ala Arg

180 185 190

His Thr Asp Asp Glu Met Thr Gly Tyr Val Ala Thr Arg Trp Tyr Arg

195 200 205

Ala Pro Glu Ile Met Leu Asn Trp Met His Tyr Asn Gln Thr Val Asp

210 215 220

Ile Trp Ser Val Gly Cys Ile Met Ala Glu Leu Leu Thr Gly Arg Thr

225 230 235 240

Leu Phe Pro Gly Thr Asp His Ile Asp Gln Leu Lys Leu Ile Leu Arg

245 250 255

Leu Val Gly Thr Pro Gly Ala Glu Leu Leu Lys Lys Ile Ser Ser Glu

260 265 270

Ser Ala Arg Asn Tyr Ile Gln Ser Leu Ala Gln Met Pro Lys Met Asn

275 280 285

Phe Ala Asn Val Phe Ile Gly Ala Asn Pro Leu Ala Val Asp Leu Leu

290 295 300

Glu Lys Met Leu Val Leu Asp Ser Asp Lys Arg Ile Thr Ala Ala Gln

305 310 315 320

Ala Leu Ala His Ala Tyr Phe Ala Gln Tyr His Asp Pro Asp Asp Glu

325 330 335

Pro Val Ala Asp Pro Tyr Asp Gln Ser Phe Glu Ser Arg Asp Leu Leu

340 345 350

Ile Asp Glu Trp Lys Ser Leu Thr Tyr Asp Glu Val Ile Ser Phe Val

355 360 365

Pro Pro Pro Leu Asp Gln Glu Glu Met Glu Ser

370 375

37

350

Human

37

Gly Asn Ala Ala Ala Ala Lys Lys Gly Ser Glu Gln Glu Ser Val Lys

1 5 10 15

Glu Phe Leu Ala Lys Ala Lys Glu Asp Phe Leu Lys Lys Trp Glu Thr

20 25 30

Pro Ser Gln Asn Thr Ala Gln Leu Asp Gln Phe Asp Arg Ile Lys Thr

35 40 45

Leu Gly Thr Gly Ser Phe Gly Arg Val Met Leu Val Lys His Lys Glu

50 55 60

Ser Gly Asn His Tyr Ala Met Lys Ile Leu Asp Lys Gln Lys Val Val

65 70 75 80

Lys Leu Lys Gln Ile Glu His Thr Leu Asn Glu Lys Arg Ile Leu Gln

85 90 95

Ala Val Asn Phe Pro Phe Leu Val Lys Leu Glu Phe Ser Phe Lys Asp

100 105 110

Asn Ser Asn Leu Tyr Met Val Met Glu Tyr Val Ala Gly Gly Glu Met

115 120 125

Phe Ser His Leu Arg Arg Ile Gly Arg Phe Ser Glu Pro His Ala Arg

130 135 140

Phe Tyr Ala Ala Gln Ile Val Leu Thr Phe Glu Tyr Leu His Ser Leu

145 150 155 160

Asp Leu Ile Tyr Arg Asp Leu Lys Pro Glu Asn Leu Leu Ile Asp Gln

165 170 175

Gln Gly Tyr Ile Gln Val Thr Asp Phe Gly Phe Ala Lys Arg Val Lys

180 185 190

Gly Arg Thr Trp Thr Leu Cys Gly Thr Pro Glu Tyr Leu Ala Pro Glu

195 200 205

Ile Ile Leu Ser Lys Gly Tyr Asn Lys Ala Val Asp Trp Trp Ala Leu

210 215 220

Gly Val Leu Ile Tyr Glu Met Ala Ala Gly Tyr Pro Pro Phe Phe Ala

225 230 235 240

Asp Gln Pro Ile Gln Ile Tyr Glu Lys Ile Val Ser Gly Lys Val Arg

245 250 255

Phe Pro Ser His Phe Ser Ser Asp Leu Lys Asp Leu Leu Arg Asn Leu

260 265 270

Leu Gln Val Asp Leu Thr Lys Arg Phe Gly Asn Leu Lys Asn Gly Val

275 280 285

Asn Asp Ile Lys Asn His Lys Trp Phe Ala Thr Thr Asp Trp Ile Ala

290 295 300

Ile Tyr Gln Arg Lys Val Glu Ala Pro Phe Ile Pro Lys Phe Lys Gly

305 310 315 320

Pro Gly Asp Thr Ser Asn Phe Asp Asp Tyr Glu Glu Glu Glu Ile Arg

325 330 335

Val Ser Ile Asn Glu Lys Cys Gly Lys Glu Phe Thr Glu Phe

340 345 350

38

422

Human

38

Met Ser Leu His Phe Leu Tyr Tyr Cys Ser Glu Pro Thr Leu Asp Val

1 5 10 15

Lys Ile Ala Phe Cys Gln Gly Phe Asp Lys Gln Val Asp Val Ser Tyr

20 25 30

Ile Ala Lys His Tyr Asn Met Ser Lys Ser Lys Val Asp Asn Gln Phe

35 40 45

Tyr Ser Val Glu Val Gly Asp Ser Thr Phe Thr Val Leu Lys Arg Tyr

50 55 60

Gln Asn Leu Lys Pro Ile Gly Ser Gly Ala Gln Gly Ile Val Cys Ala

65 70 75 80

Ala Tyr Asp Ala Val Leu Asp Arg Asn Val Ala Ile Lys Lys Leu Ser

85 90 95

Arg Pro Phe Gln Asn Gln Thr His Ala Lys Arg Ala Tyr Arg Glu Leu

100 105 110

Val Leu Met Lys Cys Val Asn His Lys Asn Ile Ile Ser Leu Leu Asn

115 120 125

Val Phe Thr Pro Gln Lys Thr Leu Glu Glu Phe Gln Asp Val Tyr Leu

130 135 140

Val Met Glu Leu Met Asp Ala Asn Leu Cys Gln Val Ile Gln Met Glu

145 150 155 160

Leu Asp His Glu Arg Met Ser Tyr Leu Leu Tyr Gln Met Leu Cys Gly

165 170 175

Ile Lys His Leu His Ser Ala Gly Ile Ile His Arg Asp Leu Lys Pro

180 185 190

Ser Asn Ile Val Val Lys Ser Asp Cys Thr Leu Lys Ile Leu Asp Phe

195 200 205

Gly Leu Ala Arg Thr Ala Gly Thr Ser Phe Met Met Thr Pro Tyr Val

210 215 220

Val Thr Arg Tyr Tyr Arg Ala Pro Glu Val Ile Leu Gly Met Gly Tyr

225 230 235 240

Lys Glu Asn Val Asp Ile Trp Ser Val Gly Cys Ile Met Gly Glu Met

245 250 255

Val Arg His Lys Ile Leu Phe Pro Gly Arg Asp Tyr Ile Asp Gln Trp

260 265 270

Asn Lys Val Ile Glu Gln Leu Gly Thr Pro Cys Pro Glu Phe Met Lys

275 280 285

Lys Leu Gln Pro Thr Val Arg Asn Tyr Val Glu Asn Arg Pro Lys Tyr

290 295 300

Ala Gly Leu Thr Phe Pro Lys Leu Phe Pro Asp Ser Leu Phe Pro Ala

305 310 315 320

Asp Ser Glu His Asn Lys Leu Lys Ala Ser Gln Ala Arg Asp Leu Leu

325 330 335

Ser Lys Met Leu Val Ile Asp Pro Ala Lys Arg Ile Ser Val Asp Asp

340 345 350

Ala Leu Gln His Pro Tyr Ile Asn Val Trp Tyr Asp Pro Ala Glu Val

355 360 365

Glu Ala Pro Pro Pro Gln Ile Tyr Asp Lys Gln Leu Asp Glu Arg Glu

370 375 380

His Thr Ile Glu Glu Trp Lys Glu Leu Ile Tyr Lys Glu Val Met Asn

385 390 395 400

Ser Glu Glu Lys Thr Lys Asn Gly Val Val Lys Gly Gln Pro Ser Pro

405 410 415

Ser Ala Gln Val Gln Gln

420

39

Val Phe Pro Ser Ser Val Phe Val Pro Asp Glu Trp Glu Val Ser Arg

1 5 10 15

Glu Lys Ile Thr Leu Leu Arg Glu Leu Gly Gln Gly Ser Phe Gly Met

20 25 30

Val Tyr Glu Gly Asn Ala Arg Asp Ile Ile Lys Gly Glu Ala Glu Thr

35 40 45

Arg Val Ala Val Lys Thr Val Asn Glu Ser Ala Ser Leu Arg Glu Arg

50 55 60

Ile Glu Phe Leu Asn Glu Ala Ser Val Met Lys Gly Phe Thr Cys His

65 70 75 80

His Val Val Arg Leu Leu Gly Val Val Ser Lys Gly Gln Pro Thr Leu

85 90 95

Val Val Met Glu Leu Met Ala His Gly Asp Leu Lys Ser Tyr Leu Arg

100 105 110

Ser Leu Arg Pro Glu Ala Glu Asn Asn Pro Gly Arg Pro Pro Pro Thr

115 120 125

Leu Gln Glu Met Ile Gln Met Ala Ala Glu Ile Ala Asp Gly Met Ala

130 135 140

Tyr Leu Asn Ala Lys Lys Phe Val His Arg Asp Leu Ala Ala Arg Asn

145 150 155 160

Cys Met Val Ala His Asp Phe Thr Val Lys Ile Gly Asp Phe Gly Met

165 170 175

Thr Arg Asp Ile Xaa Glu Thr Asp Xaa Xaa Arg Lys Gly Gly Lys Gly

180 185 190

Leu Leu Pro Val Arg Trp Met Ala Pro Glu Ser Leu Lys Asp Gly Val

195 200 205

Phe Thr Thr Ser Ser Asp Met Trp Ser Phe Gly Val Val Leu Trp Glu

210 215 220

Ile Thr Ser Leu Ala Glu Gln Pro Tyr Gln Gly Leu Ser Asn Glu Gln

225 230 235 240

Val Leu Lys Phe Val Met Asp Gly Gly Tyr Leu Asp Gln Pro Asp Asn

245 250 255

Cys Pro Glu Arg Val Thr Asp Leu Met Arg Met Cys Trp Gln Phe Asn

260 265 270

Pro Lys Met Arg Pro Thr Phe Leu Glu Ile Val Asn Leu Leu Lys Asp

275 280 285

Asp Leu His Pro Ser Phe Pro Glu Val Ser Phe Phe His Ser Glu Glu

290 295 300

Asn Lys

305

40

306

Human

40

Val Phe Pro Ser Ser Val Phe Val Pro Asp Glu Trp Glu Val Ser Arg

1 5 10 15

Glu Lys Ile Thr Leu Leu Arg Glu Leu Gly Gln Gly Ser Phe Gly Met

20 25 30

Val Tyr Glu Gly Asn Ala Arg Asp Ile Ile Lys Gly Glu Ala Glu Thr

35 40 45

Arg Val Ala Val Lys Thr Val Asn Glu Ser Ala Ser Leu Arg Glu Arg

50 55 60

Ile Glu Phe Leu Asn Glu Ala Ser Val Met Lys Gly Phe Thr Cys His

65 70 75 80

His Val Val Arg Leu Leu Gly Val Val Ser Lys Gly Gln Pro Thr Leu

85 90 95

Val Val Met Glu Leu Met Ala His Gly Asp Leu Lys Ser Tyr Leu Arg

100 105 110

Ser Leu Arg Pro Glu Ala Glu Asn Asn Pro Gly Arg Pro Pro Pro Thr

115 120 125

Leu Gln Glu Met Ile Gln Met Ala Ala Glu Ile Ala Asp Gly Met Ala

130 135 140

Tyr Leu Asn Ala Lys Lys Phe Val His Arg Asp Leu Ala Ala Arg Asn

145 150 155 160

Cys Met Val Ala His Asp Phe Thr Val Lys Ile Gly Asp Phe Gly Met

165 170 175

Thr Arg Asp Ile Tyr Glu Thr Asp Tyr Tyr Arg Lys Gly Gly Lys Gly

180 185 190

Leu Leu Pro Val Arg Trp Met Ala Pro Glu Ser Leu Lys Asp Gly Val

195 200 205

Phe Thr Thr Ser Ser Asp Met Trp Ser Phe Gly Val Val Leu Trp Glu

210 215 220

Ile Thr Ser Leu Ala Glu Gln Pro Tyr Gln Gly Leu Ser Asn Glu Gln

225 230 235 240

Val Leu Lys Phe Val Met Asp Gly Gly Tyr Leu Asp Gln Pro Asp Asn

245 250 255

Cys Pro Glu Arg Val Thr Asp Leu Met Arg Met Cys Trp Gln Phe Asn

260 265 270

Pro Lys Met Arg Pro Thr Phe Leu Glu Ile Val Asn Leu Leu Lys Asp

275 280 285

Asp Leu His Pro Ser Phe Pro Glu Val Ser Phe Phe His Ser Glu Glu

290 295 300

Asn Lys

305

41

Lys Pro Trp Trp Glu Asp Glu Trp Glu Val Pro Arg Glu Thr Leu Lys

1 5 10 15

Leu Val Glu Arg Leu Gly Ala Gly Gln Phe Gly Glu Val Trp Met Gly

20 25 30

Tyr Tyr Asn Gly His Thr Lys Val Ala Val Lys Ser Leu Lys Gln Gly

35 40 45

Ser Met Ser Pro Asp Ala Phe Leu Ala Glu Ala Asn Leu Met Lys Gln

50 55 60

Leu Gln His Gln Arg Leu Val Arg Leu Tyr Ala Val Val Thr Gln Glu

65 70 75 80

Pro Ile Tyr Ile Ile Thr Glu Tyr Met Glu Asn Gly Ser Leu Val Asp

85 90 95

Phe Leu Lys Thr Pro Ser Gly Ile Lys Leu Thr Ile Asn Lys Leu Leu

100 105 110

Asp Met Ala Ala Gln Ile Ala Glu Gly Met Ala Phe Ile Glu Glu Arg

115 120 125

Asn Tyr Ile His Arg Asp Leu Arg Ala Ala Asn Ile Leu Val Ser Asp

130 135 140

Thr Leu Ser Cys Lys Ile Ala Asp Phe Gly Leu Ala Arg Leu Ile Glu

145 150 155 160

Asp Asn Glu Xaa Thr Ala Arg Glu Gly Ala Lys Phe Pro Ile Lys Trp

165 170 175

Thr Ala Pro Glu Ala Ile Asn Tyr Gly Thr Phe Thr Ile Lys Ser Asp

180 185 190

Val Trp Ser Phe Gly Ile Leu Leu Thr Glu Ile Val Thr His Gly Arg

195 200 205

Ile Pro Tyr Pro Gly Met Thr Asn Pro Glu Val Ile Gln Asn Leu Glu

210 215 220

Arg Gly Tyr Arg Met Val Arg Pro Asp Asn Cys Pro Glu Glu Leu Tyr

225 230 235 240

Gln Leu Met Arg Leu Cys Trp Lys Glu Arg Pro Glu Asp Arg Pro Thr

245 250 255

Phe Asp Tyr Leu Arg Ser Val Leu Glu Asp Phe Phe Thr Ala Thr

260 265 270