CA2306455A1 - Human receptor proteins; related reagents and methods - Google Patents

Abstract

Nucleic acids encoding mammalian, e.g., human receptors, purified receptor proteins and fragments thereof. Antibodies, both polyclonal and monoclonal, are also provided. Methods of using the compositions for both diagnostic and therapeutic utilities are provided.

Description

HUMAN RECEPTOR PROTEINS; RELATED REAGENTS AND METHODS
FIELD OF THE INVENTION
The present invention relates to compositions and methods for affecting mammalian physiology, including, e.g., morphogenesis or immune system function. In particular, it provides nucleic acids, proteins, and antibodies, e.g., which regulate development and/or the immune system along with related reagents and methods.
Diagnostic and therapeutic uses of these materials are also disclosed.
BACKGROUND OF THE INVENTION
Recombinant DNA technology refers generally to techniques of integrating genetic information from a donor source into vectors for subsequent processing, such as through introduction into a host, whereby the transf erred genetic information is copied and/or expressed in the new environment. Commonly, the genetic information exists in the form of complementary DNA
(cDNA) derived from messenger RNA (mRNA) coding for a desired polypeptide product. The carrier is frequently a plasmid having the capacity to incorporate cDNA for later replication and/or expression in a host and, in some cases, actually to control expression of the cDNA and thereby direct synthesis of the encoded product in the host.
For some time, it has been known that the mammalian immune response is based on a series of complex cellular interactions, called the "immune network". Recent research has provided new insights into the inner workings of this network. While it remains clear that much of the immune response does, in fact, revolve around the network-like interactions of lymphocytes, macrophages, granulocytes, and other cells, immunologists now generally hold the opinion that soluble proteins, known as lymphokines, cytokines, or monokines, play critical roles in controlling these cellular interactions. Thus, there is considerable interest in the isolation, characterization, and mechanisms of action of cell modulatory factors, an understanding of which will lead to significant advancements in the diagnosis and therapy of numerous medical abnormalities, e.g., immune system disorders.
Lymphokines apparently mediate cellular activities in a variety of ways. They have been shown to support the proliferation, growth, and/or differentiation of pluripotential hematopoietic stem cells into vast numbers of progenitors comprising diverse cellular lineages which make up a complex immune system. Proper and balanced interactions between the cellular components are necessary for a healthy immune response. The different cellular lineages often respond in a different manner when lymphokines are administered in conjunction with other agents.
Cell lineages especially important to the immune response include two classes of lymphocytes: B-cells, which can produce and secrete immunoglobulins (proteins with the capability of recognizing and binding to foreign matter to effect its removal), and T-cells of various subsets that secrete lymphokines and induce or suppress the B-cells and various other cells (including other T-cells) making up the immune network. These lymphocytes interact with many other cell types.
Another important cell lineage is the mast cell (which has not been positively identified in all mammalian species), which is a granule-containing connective tissue cell located proximal to capillaries throughout the body. These cells are found in especially high concentrations in the lungs, skin, and gastrointestinal and genitourinary tracts. Mast cells play a central role in allergy-related disorders, particularly anaphylaxis as follows: when selected antigens crosslink one class of immunoglobulins bound to receptors on the mast cell surface, the mast cell degranulates and releases mediators, e.g., histamine, serotonin, heparin, and prostaglandins, which cause allergic reactions, e.g., anaphylaxis.

3 _ Research to better understand and treat various immune disorders has been hampered by the general inability to maintain cells of the immune system in vitro. Immunologists have discovered that culturing many of these cells can be accomplished through the use of T-cell and other cell supernatants, which contain various growth factors, including many of the lymphokines.
The interleukin-1 family of proteins includes the IL-loc, the IL-1(3, the IL-1RA, and recently the IL-1'y (also designated Interferon-Gamma Inducing Factor, IGIF).
This related family of genes has been implicated in a broad range of biological functions. See Dinarello (1994) FASEB J. 8:1314-1325; Dinarello (1991) Blood 77:1627-1652; and Okamura, et al. (1995) ure 378:88-91.
From the foregoing, it is evident that the discovery and development of new soluble proteins and their receptors, including ones similar to lymphokines, should contribute to new therapies. A number of degenerative or abnormal conditions directly or indirectly involve development, differentiation, or function, e.g., of the immune system and/or hematopoietic cells. In particular, the discovery and understanding of novel receptors for lymphokine-like molecules which enhance or potentiate the beneficial activities of other lymphokines, would be highly advantageous. The present invention provides new receptors for ligands exhibiting similarity to interleukin-1 like compositions and related compounds, and methods for their use.
SUMMARY OF THE INVENTION
The present invention is directed to novel receptors related to IL-1 receptors and their biological activities. These receptors, e.g., primate or rodent, are designated IL-1 receptor like molecular structures, IL-1 Receptor DNAX designation 8(IL-1RD8), IL-1 Receptor DNAX designation 9(IL-1RD9) and IL-1 Receptor DNAX
designation 10(IL-1RD10). The invention includes nucleic acids coding for the polypeptides themselves and methods 4 _ for their production and use. The nucleic acids of the invention are characterized, in part, by their homology to cloned complementary DNA (cDNA) sequences enclosed herein.
In certain embodiments, the invention provides a composition of matter selected from the group of: an isolated or recombinant IL-1RD8 polypeptide comprising a segment of at least 12 contiguous amino acids of SEQ ID
NO: 2 or 4, a natural sequence IL-1RD8 polypeptide comprising SEQ ID NO: 2 or 4, a fusion protein comprising IL-1RD8 sequence; an isolated or recombinant IL-1RD9 polypeptide comprising at least 12 contiguous amino acids of SEQ ID NO: 6, 8, 10, 12, 14, or 16; a natural sequence IL-1RD9 comprising SEQ ID N0: 6, 8, 10, 12, 14, or 16; a fusion protein comprising IL-1RD9 sequence; an isolated or recombinant IL-1RD10 polypeptide comprising at least 12 contiguous amino acids of SEQ ID NO: 18 or 20; a natural sequence IL-1RD10 comprising SEQ ID NO: 18 or 20;
and a fusion protein comprising IL-1RD10 sequence. In various embodiments, the recombinant or isolated polypeptide comprises a segment identical to a corresponding portion of an IL-1RD8, as described, wherein: the number of contiguous amino acid residues is:
at least 17 amino acids; at least 21 amino acids; or at least 25 amino acids; or to a corresponding portion of an IL-1RD9, as described, wherein the number of identical contiguous amino acid residues is: at least 17 amino acids; at least 21 amino acids; or at least 25 amino acids; or of an IL-1RD10, as described, wherein the number of identical contiguous amino acid residues is: at least 17 amino acids; at least 21 amino acids; or at least 25 amino acids.
In polypeptide embodiments, the invention provides a composition of matter wherein the IL-1RD8 comprises a mature sequence shown in SEQ ID NO: 2 or 4; an IL-1RD9 that comprises a mature sequence shown in SEQ ID NO: 6, 8, 10, 12, 14 or 16; an IL-1RD10 that comprises a mature sequence shown in SEQ ID NO: 18 or 20; or the IL-1RD8, IL-1RD9, or IL-1RD10 polypeptide: is from a warm blooded animal, e.g., a primate, such as a human; comprises at least one polypeptide segment of SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 18, or 20; exhibits a plurality of portions having segments identical to specific sequence 5 identifiers; is a natural allelic variant of a primate IL-1RD8; a primate or rodent IL-1RD9; or a primate IL-1RD10; has a length at least about 30 amino acids;
exhibits at least two non-overlapping epitopes that are specific for: a primate IL-1RD8, a primate or rodent IL-1RD9, or primate IL-1RD10; exhibits a sequence identity over a length of at least about 20 amino acids to: a primate IL-1RD8, a primate or rodent IL-1RD9, or a primate IL-1RD10; has a molecular weight of at least 100 kD with natural glycosylation; is a synthetic polypeptide; is attached to a solid substrate; is conjugated to another chemical moiety; is a 5-fold or less substitution from natural sequence; or is a deletion or insertion variant from a natural sequence. Certain preferred embodiments include compositions comprising: a sterile IL-1RD8, IL-1RD9, or IL-1RD10 polypeptide; or the IL-1RD8, IL-1RD9, or IL-1RD10 polypeptide and a carrier, wherein the carrier is: an aqueous compound, including water, saline, and/or buffer; and/or formulated for oral, rectal, nasal, topical, or parenteral administration; a sterile IL-1RD8, IL-1RD9, or IL-1RD10 polypeptide; or the IL-1RD8, IL-1RD9, or IL-1RD10 polypeptide, as described, and a carrier, wherein the carrier is: an aqueous compound, including water, saline, and/or buffer; and/or formulated for oral, rectal, nasal, topical, or parenteral administration.
Certain fusion proteins are provided, e.g., comprising: mature polypeptide sequence shown in SEQ ID
NO: 2, 4, 6, 8, 10, 12, 14, 16, 18 or 20; a detection or purification tag, including a FLAG, His6, or Ig sequence;
or sequence of another receptor protein. Kit embodiments include a kit comprising such a polypeptide, and: a compartment comprising the polypeptide; and/or instructions for use or disposal of reagents in the kit.

WO 99/19480 ~ PCT/US98/20939 In binding compound embodiments, the invention provides a binding compound comprising an antigen binding -site from an antibody, which specifically binds to a natural: IL-1RD8, IL-1RD9, or IL-1RD10 polypeptide, wherein: the polypeptide is a primate or rodent protein;
the binding compound is an Fv, Fab, or Fab2 fragment; the binding compound is conjugated to another chemical moiety; or the antibody: is raised to a polypeptide sequence of a mature polypeptide comprising a sequence sequence shown in SEQ ID NO: 2, 4, 6, 8, 10, 12, 14 , 16, 18 or 20; is raised to a mature primate or rodent IL-1RD8; is raised to a purified human IL-1RD8; is raised to a purified mouse IL-1RD9; is immunoselected; is a polyclonal antibody; binds to a denatured IL-1RD8, IL-1RD9, or IL-1RD10; exhibits a Kd to antigen of at least 30 ~,tM; is attached to a solid substrate, including a bead or plastic membrane; is in a sterile composition; or is detestably labeled, including a radioactive or fluorescent label; IL-1RD9 protein, wherein: the polypeptide is a primate or rodent protein; the binding compound is an Fv, Fab, or Fab2 fragment; the binding compound is conjugated to another chemical moiety; or the antibody: is raised against a polypeptide sequence of a mature polypeptide comprising a sequence sequence shown in SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 18 or 20; is raised against a mature primate IL-1RD9; is raised to a purified human IL-1RD9; is immunoselected; is a polyclonal antibody; binds to a denatured IL-1RD9;
exhibits a Kd to antigen of at least 30 ~1M; is attached to a solid substrate, including a bead or plastic membrane; is in a sterile composition; or is detestably labeled, including a radioactive or fluorescent label;
IL-1RD10 protein, wherein: the polypeptide is a primate or rodent protein; the binding compound is an Fv, Fab, or Fab2 fragment; the binding compound is conjugated to another chemical moiety; or the antibody: is raised against a polypeptide sequence of a mature polypeptide comprising a sequence sequence shown in SEQ ID N0: 2, 4, 6, 8, 10, 12, 14, 16, 18 or 20; is raised against a WO 99/19480 PCT/US98l20939 7 t mature primate IL-1RD10; is raised to a purified human IL-1RD10; is immunoselected; is a polyclonal antibody;
binds to a denatured IL-1RD10; exhibits a Kd to antigen of at least 30 E1M; is attached to a solid substrate, including a bead or plastic membrane; is in a sterile composition; or is detectably labeled, including a radioactive or fluorescent label. Kits are provided, e.g., those comprising the binding compound, and: a compartment comprising the binding compound; and/or instructions for use or disposal of reagents in the kit.
Preferably, the kit is capable of making a qualitative or quantitative analysis.
Other embodiments include a composition comprising:
a sterile binding compound, or the binding compound and a carrier, wherein the carrier is: an aqueous compound, including water, saline, and/or buffer; and/or formulated for oral, rectal, nasal, topical, or parenteral administration.
Nucleic acid embodiments include an isolated or recombinant nucleic acid encoding a polypeptide or fusion protein, wherein: the IL-1RD8, IL-1RD9, or IL-1RD10 is from a mammal; said nucleic acid: encodes an antigenic polypeptide sequence sequence shown in SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 18 or 20; encodes a plurality of antigenic polypeptide sequences sequence shown in SEQ ID
NO: 2, 4, 6, 8, 10, 12, 14, 16, 18 or 20; exhibits at least about 30 nucleotides to a natural cDNA encoding the segment; is an expression vector; further comprises an origin of replication; is from a natural source;
comprises a detectable label; comprises synthetic nucleotide sequence; is less than 6 kb, preferably less than 3 kb; is from a mammal, including a primate;
comprises a natural full length coding sequence; is a hybridization probe for a gene encoding said IL-1RD8, IL-1RD9, or IL-1RD10; comprises a plurality of nonoverlapping segments of at least 15, 18, 21, or 25 nucleotides shown in SEQ ID NO: 1, 3, 5, 7, 9 11, 13, 15, 17, 19; or is a PCR primer, PCR product, or mutagenesis primer. The invention further provides a cell comprising such a recombinant nucleic acid, e.g., where the cell is:
a prokaryotic cell; a eukaryotic cell; a bacterial cell;
a yeast cell; an insect cell; a mammalian cell; a mouse cell; a primate cell; or a human cell. Certain kit embodiments include a comprising the nucleic acid, and: a compartment comprising the nucleic acid; a compartment further comprising: a primate IL-1RD8, a primate or rodent IL-1RD9, or a primate IL-1RD10 polypeptide; and/or instructions for use or disposal of reagents in the kit.
Preferably, the kit is capable of making a qualitative or quantitative analysis.
In other nucleic acid embodiments, the nucleic acid is one which: hybridizes under wash conditions of 40° C
and less than 2M salt to either SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 17, or 19; or exhibits identity over a stretch of at least about 30 nucleotides to a primate IL-1RD8, a primate or rodent IL-1RD9, or a primate IL-1RD10.
In various preferred embodiments: the wash conditions are: at 45° C and/or 500 mM salt; at 55° C and/or 150 mM
salt; or the stretch is at least 55 nucleotides; or at least 75 nucleotides.
Methods of modulating physiology or development of a cell or tissue culture cells are provided, e.g., comprising contacting the cell with an agonist or antagonist of a primate IL-1RD8, a primate or rodent IL-1RD9, or a primate IL-1RD10. Preferably, the cell is transformed with a nucleic acid encoding either IL-1RD8, IL-2RD9, or IL-1RD10, and another IL-1R.
DETAILED DESCRIPTION OF THE INVENTION
I. General The present invention provides the amino acid sequence and DNA sequence of mammalian, herein, e.g., primate and rodent IL-1 receptor-like molecules; these molecules IL-1 Receptor DNAX designation 8(IL-1RD8), IL-1 Receptor DNAX designation 9(IL-1RD9) and IL-1 Receptor."
DNAX designation 10(IL-1RD10) having particular defined properties, both structural and/or biological. These embodiments increase the number of members of the human IL-1 receptor-like family from 7 to at least 10. These receptors have been numbered internally as DNAX
designations D1, D2, D3, D4, D5, D6, and now D8, D9, and D10, and are referred to as 'IL-1RD1 through D10. Various cDNAs encoding these molecules were obtained from primate, e.g., human, or rodent, e.g., mouse, cDNA
sequence libraries. Other primate, rodent, or other mammalian counterparts would also be desired.
Some of the standard methods applicable are described or referenced, e.g., in Maniatis, et al. (1982) Molecular Cloninct A Laboratory Manual, Cold Spring Harbor Laboratory, Cold Spring Harbor Press; Sambrook, et al. (1989) molecular Clonincr: A Laboratory Manual, (2d.
ed.), vols. 1-3, CSH Press, NY; Ausubel, et al: Biolocrv, Greene Publishing Associates, Brooklyn, NY; or Ausubel, et al. (1987 and periodic supplements) Current Protocols in Molecular Bioloav, Greene/Wiley, New York; each of which is incorporated herein by reference.
A partial nucleotide and corresponding amino acid sequence of a human IL-1RD8 coding segment is shown in SEQ ID NO: 1 and 2, respectively. Supplemental human IL-1RD8 nucleotide and corresponding sequence is provided in SEQ ID NO: 3 and 4, respectively.
Similarly for primate IL-1RD9, partial nucleotides (SEQ ID NO: 5) and corresponding amino acid sequences (SEQ ID NO: 6) of a primate IL-1RD9 coding segment are provided. Supplemental primate IL-1RD9 is provided in SEQ ID NO: 7, 8, 9, and 10. Rodent embodiments of IL-1RD9 are provided in SEQ ID NO: 11, 12, with supplemental IL-1RD9 rodent sequence in SEQ ID N0: 13, 14, 15, and 16.
For an embodiment of human IL-1RD10, a partial nucleotide and corresponding partial amino acid sequence are provided in SEQ ID N0: 17 and 18, respectively, with supplemental human IL-1RD10 nucleotide and corresponding partial amino acid sequence provided in SEQ ID NO: 19 and 20, respectively.
Some sequences provided lack some portions of these receptors, as suggested by alignment of sequences shown in Tables 1-4). Note the alignment of IL-1RD10 with IL-1RD8 and D3s, which are alpha type receptor subunits.
Table 4 exhibits alignment of primate and rodent IL-1RD9.
It is to be understood that this invention is not limited to the particular methods, compositions and 5 receptors specifically embodied herein, as such methods, compositions and receptors may, of course, vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to limit the scope of the 10 present invention which is only limited by the appended claims.
Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by a person of ordinary skill in the art to which this invention belongs. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, suitable methods and materials are described below. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety including all figures, graphs, and drawings.
Table 1 Alignment of the extracellular domains of various IL-lRs.
hIL-1RD10 is SEQ ID NO: 20; hIL-1RD8 is SEQ ID NO:
3; mIL-1RD3 is GenBank X85999: hIL-1RD6 is GenBank U49065; rIL-1RD6is GenBank 3 U49066: mIL-1RD4 hIL-1RD4 O is GenBank Y07519 is and GenBank D13695;

GenBank D12763; hIL-1RD2 is GenBank X59770; mIL-1RD2is GenBank X59769; hIL-1RD5 GenBank U43672; mIL-1RD5 is is GenBank U43673;

mIL-1RD1 is GenBank M20658, M29752; hIL-1RD1 is GenBank X16896;

cIL-1RD1 is GenBank 86325; and hFGR4 is GenBank 55. Other 3 species counterparts 5 may be obtained from public sequence databases.

mIL-1RD3 .......... ......MGLL WYLMSLSFYG ILQSHASERCDDWLDTMR..

hIL-1RD6 .........- .........M WSLLLCGLSI ALPLSVTADGCKDIFMKN..

40 rIL-1RD6 .......... .--....MGM PPLLFCWVSF VLPLFVAAGNCTDVYMHH..

mIL-1RD4 .......... ........MI DRQRMGLWAL AILTLPMYLTVTEGSKSS..

hIL-1RD4 .......... ........MG FWILAILTIL MYSTAAKFSKQS........

hIL-1RD2 .......... ....MLRLYV LVMGVSAFTL QPAAHTGAARSCRFRGRHYK

mIL-1RD2 MFILLVLVTG VSAFTTPTW HTGKVSESPI TSEKPTVHGDNCQFRGREFK

45 hIL-1RD10 .................... .......... .....................

hIL-1RD5 .......... .....MNCRE LPLTLWVLIS VSTAESCTSRPHITVVE...

mIL-1RD5 .......... .....MHHEE LILTLCILIV KSASKSCIHRSQIHWE...

mIL-1RD1 .......... .....MENMK VLLGLICLMV PLLSLEIDVCTEYPNQIVLF

hIL-1RD1 .......... .....-..MK VLLRLICFIA LLISSLEADKCKEREEKIIL

5 cIL-1RD1 .......... .....MHKMT STFLLIGHLI LLIPLFSAEECVICNYFVLV
O

hIL-1RD8 .........M KPPFLLALW CSWSTNLKM VSKRNSVDGCIDWSVDLKTY

hFGR4 ...MRLLLAL LGVLLSVPGP PVLSLEASEE VELEPCLAPS LEQQEQELTV
mIL-1RD3 QIQVFEDEPA RIKCPLFEHFLKYNYSTAHSSGLTLLWYWTRQDRDLEEPI

hIL-1RD6 .EILSASQPF AFNCTFPPI.........TSGEVSVTWYKN....SSKIPV

rIL-1RD6 .EMISEGQPF PFNCTYPPV.........TNGAVNLTWHRT....PSKSPI

mIL-1RD4 ..WGLENEAL IVRCPQRG...........RSTYPVEWYYS....DTNESI

hIL-1RD4 ..WGLENEAL IVRCPRQG...........KPSYTVDWYYS. ,..Q~'NKSI

hIL-1RD2 REFRLEGEPV ALRCPQVPYWLWA....SVSPRINLTWHKN.,..DSARTV

mIL-1RD2 SELRLEGEPV VLRCPLAPHSDIS.....SSSHSFLTWSKL....DSSQLI

hIL-1RD10 .......... ........................................

hIL-1RD5 .....GEPFY LKHCSCSLAH........EIETTTKSWYKS...SGSQEHV

mIL-1RD5 .....GEPFY LKPCGISAPV.......HRNETATMRWFKG...SASHEYR

mIL-1RD1 LSV...NEID IRKCPLTPN.........KMHGDTIIWYKN....DSKTPI

hIL-1RD1 VSS..ANEID VRPCPLNPN..........EHKGTITWYKD....DSKTPV

cIL-1RD1 ......GEPT AISCPVITL.......PMLHSDYNLTWYRN....GSNMPI

hIL-1RD8 ..MALAGEPV RVKCALFYSYIRTNYSTAQSTGLRLMWYKN..KGDLEEPI

hFGR4 ....ALGQPV RLCCGRAERGG............ " ....GSRLAP
HWYKE

mIL-1RD3 NFRLP.ENRI SKEKDVLWFR PTLLNDTGNY TCMLRNTTYC SKVAE'PLEW
2 O hIL-1RD6 SKII..QSRIHQDETWILFLPMEWGDSGVYQCVIKGRDSCHRIHVNLTVF

rIL-1RD6 SINR..HVRIHQDQSWILFLPLALEDSGIYQCVIKDAHSCYRIAINLTVF

mIL-1RD4 PTQK,.RNRIFVSRDRLKFLPARVEDSGIYACVIRSPNLNKTGYLNVTIH

hIL-1RD4 PTQE..RNRVFASGQLLKFLPAEVADSGIYTCIVRSPTFNRTGYANVTIY

hIL-1RD2 PGEE..ETRMWAQDGALWLLPALQEDSGTYVCTTRNASYCDKMSIELRVF

2 5 mIL-1RD2 PRDEP...RMWVKGNILWILPAVQQDSGTYICTFRNASHCEQMSVELKVF

hIL-1RD10 ..................................................

hIL-1RD5 ELNPRSSSRIALHDCVLEFWPVELNDTGSYFFQMKN..YTQKWKLNVIRR

mIL-1RD5 ELNNRSSPRVTFHDHTLEFWPVEMEDEGTYISQVGN..DRRNWTLNVTKR

mIL-1RD1 SADR..DSRIHQQNEHLWFVPAKVEDSGYYYCIVRNSTYCLKTKVTVTVL

3 O hIL-1RD1 STEQ..ASRIHQHKEKLWFVPAKVEDSGHYYCVVRNSSYCLRIKISAKFV

cIL-1RD1 TTER..RARIHQRKGLLWFIPAALEDSGLYECEVRSLNRSKQKIINLKVF

hIL-1RD8 IFS...EVRMSKEEDSIWFHSAEAQDSGFYTCVLRNSTYCMKVSMSLTVA

hFGR4 AG......RVRGWRGRLEIASFLPEDAGRYLCLARGSMIVLQNLTLITGD

3 mIL-1RD3 QK.................. .......DSCFNSAMRFPVHKMYIEHGIHK

hIL-1RD6 EK.................. .HWCDTSIGGLP.NLSDEYKQILHLGKDDS

rIL-1RD6 RK.................. .HWCDSSNEESSINSSDEYQQWLPIGKSGS

mIL-1RD4 KK.................. .....PPSCN.IPDY.LMYSTVRGSDKNFK

hIL-1RD4 KK.................. .....QSDCN.VPDY.LMYSTVSGSEKNSK

4O hIL-1RD2 EN.................. .......TDAFLPFI..SYPQILTLSTSGV

mIL-1RD2 KN.................. .......TEASLPHV..SYLQISALSTTGL

hIL-1RD10 .................... ..............................

hIL-1RD5 NK.................. .......HSCFTERQ..VTSKIVEVKKFFQ

mIL-1RD5 NK.................. .......HSCFSDKL..VTSRDVEVNKSLH

45 mIL-1RD1 EN.................. .....DPGIC.YSTQ.ATFPQRLHIAGDGS

hIL-1RD1 EN.................. .....EPNLC.YNAQ.AIFKQKLPVAGDGG

cIL-1RD1 KN.................. .....DNGLC.FNGE.MKYDQIVKSANAGK

hIL-1RD8 EN.................. .....ESGLC.YNSR.IRYLEKSEVTKRKE

hFGR4 SLTSSNDDEDPKSHRDPSNR HSYPQQAPYWTHPQRMEKKLHAVPAGNTVK

mIL-1RD3 ITCPNVDGYFP.SSVKPSVT WYKGCTEIVDFHN...VLPEGMNLSFFIPL

hIL-1RD6 LTCHLHFPKS...CVLGPIK WYKDCNEIKGE......RFTVLETRLLVSN

rIL-1RD6 LTCHLYFPES...CVLDSIK WYKGCEEIKV5.....KKFCPTGTKLLVNN

mIL-1RD4 ITCPTIDLY...,NWTAPVQ WFKNCKALQEP......RFRAHRSYLFIDN

55 hIL-1RD4 IYCPTIDLY....NWTAPLE WFKNCQALQGS......RYRAHKSFLVIDN

hIL-1RD2 LVCPDLSEFTR.DKTDVKIQ WYKDSLLLDKDNEK..FLSVRGTTHLLVHD

mIL-1RD2 LVCPDLKEFIS.SNADGKIQ WYKGAILLDKGNKE..FLSAGDPTRLLISN

hIL-1RD10 .................... ..............................

hIL-1RD5 ITCENSYYQ....TLVNSTS LYKNCKKLLLENN....KNPTIKKNAEF:.

60 mIL-1RD5 ITCKNPNYE....ELIQDTW LYKNCKEISKTPRI...LKDAEFGDAEF..

mIL-1RD1 LVCPYVSYFKDENNELPEVQ WYKNCKPLLLDN....VSFFGVKDKLLVRN

hIL-1RD1 LVCPYMEFFKNENNELPKLQ WYKDCKPLLLDN....IHFSGVKDRLIVMN

cIL-1RD1 IICPDLENFKDEDNINPEIH WYKECKSGFLEDKR..LVLAEGENAILILN

hIL-1RD8 ISCPDMDDFKKSD.QEPDW WYKECKPKMWR.....SIIIQKGNALLIQE

hFGR4 FRCPAAG... ...NPTPTIR WLKDGQAFHG ENRIGGIRLR HQHWSLVMES
mIL-1RD3 VSNN..GNYT CVVTYPENGR LFHLTRTVTV KWGS.PKDA LPPQIYSPND
hIL-1RD6 VSAEDRGNYA CQAILTHSGK QYEVLNGITV SITERAGYGG SVP.KIIYPK
rIL-1RD6 IDVEDSGSYA CSARLTHLGR IFTVRNYIAV NTKE.VGSGG RIP.NITYPK
mIL-1RD4 VTHDDEGDYT CQFTHAENGT NYIVTATRSF TVE.EKGFS. MFPVITNPPY
hIL-1RD4 VMTEDAGDYT CKFIHNENGA NYSVTATRSF TVKDEQGFS. LF,~V~GAPAQ
hIL-1RD2 VALEDAGYYR CVLTFAHEGQ QYNITRSIEL RIKKK..KEE TIPVIISP..
mIL-1RD2 TSMDDAGYYR CVMTFTYNGQ EYNITRNIEL RVKGT..TTE PIPVIISP..
1O hIL-1RD10 ...EFG..TS CEL..KYGGF V..VRRTTEL TVTAPLTDKP PKLLYPMESK
hIL-1RD5 ...EDQGYYS CVHFLHHNGK LFNITKTFNI TIVED..RSN IVPVLLGP.K
mIL-1RD5 ...GDEGYYS CVFSVHHNGT RYNITKTVNI TVIEG..RSK VTPAILGP.K
mIL-1RD1 VAEEHRGDYI CRMSYTFRGK QYPVIRVIQF ITIDE..NKR DRPVILSP.R
hIL-1RD1 VAEKHRGNYT CHASYTYLGK QYPITRVIEF ITLEE..NKP TRPVIVSP.A
15 cIL-1RD1 VTIQDKGNYT CRMVYTYMGK QYNVSRTMNL EVKES..PLK MRPEFIYP.N
hIL-1RD8 VQEEDGGNYT CEL..KYEGK L..VRRTTEL KVTALLTDKP PKPLFPMENQ
hFGR4 WPSDRGTYT CLVENAVGSI RYNYLLDVLE RSPH..RPIL QAGLPANTT.
mIL-1RD3 RVVYEKEPGE ELVIPCKWF SFIMD.SHNE VWWTIDGKKP .DDVTVDITI
2 O hIL-1RD6 NHSIEVQLGT TLIVDCNVTD TK..D.NTNL RCWRVNNTLV DDYYDESKRI
rIL-1RD6 NNSIEVQLGS TLIVDCNITD TK..E.NTNL RCWRVNNTLV DDYYNDFKRI
mIL-1RD4 NHTMEVEIGK PASIACSACF GKGSH.FLAD VLWQINKTW GNFGEARIQE
hIL-1RD4 NEIKEVEIGK NANLTCSACF GKGTQ.FLAA VLWQLNGTKI TDFGEPRIQQ
hIL-1RD2 LKTISASLGS RLTIPCKVFL GTGTP.LTTM LWWTANDTHI .ESAYPGGRV
25 mIL-1RD2 LETIPASLGS RLIVPCKVFL GTGTS.SNTI VWWLANSTFI .SAAYPRGRV
hIL-1RD10 LTIQETQLGD SANLTCRAFF GYSGD.VSPL IYWMKGEKFI EDLDENRVWE
hIL-1RD5 LNHVAVELGK NVRLNCSALL N.....EEDV IYWMFGEENG ...SDPNIHE
mIL-1RD5 CEKVGVELGK DVELNCSASL N.....KDDL FYWSIRKEDS ...SDPNVQE
mIL-1RD1 NETIEADPGS MIQLICNVTG Q.....FSDL VYWKWNGSEI .EVJNDPFLAE
3 O hIL-1RD1 NETMEVDLGS QIQLICNVTG Q.....LSDI AYWKWNGSVI .DEDDPVLGE
cIL-1RD1 NNTIEVELGS HVVMECNVSS GV....YGLL PYWQVNDEDV .DSFDSTYRE
hIL-1RD8 PSVIDVQLGK PLNIPCKAFF GFSGE.SGPM IYWMKGEKFI .EELAGHIRE
hFGR4 .....AWGS DVELLCKVYS DA...QPHIQ ..WLKHIVIN GSSFGA..DG
3 mIL-1RD3 NESVSYSSTED..ETRTQILSIKKVTPEDLRRNWCHARNTKGEAEQAAK

hIL-1RD6 REGVETHVSFREHNLYTVNITFLEVKMEDYGLPFMCHAG....VSTAYII

rIL-1RD6 QEGIETNLSLRNHILYTVNITFLEVKMEDYGHPFTCHAA....VSAAYII

mIL-1RD4 EEGRNESSSND.MDCLTSVLRITGVTEKDLSLEYDCLALNLHGMIRHTIR

hIL-1RD4 EEGQNQSFSNG.LACLDMVLRIADVKEEDLLLQYDCLALNLHGLRRHTVR

4O hIL-1RD2 TEGPRQEYSENNENYIEVPLIFDPVTREDLHMDFKCWHNTLSFQTLRTT

mIL-1RD2 TEGLHHQYSENDENYVEVSLIFDPVTREDLHTDFKCVASNPRSSQSLHTT

hIL-1RD10 SDIRILKEHLG.EQEVSISLIVDSVEEGDL.GNYSCYVENGNGRRHASVL

hIL-1RD5 EKEMRIMTPEG.KWHASKVLRIENIGESNLNVLYNCTVASTGGTDTKSFI

mIL-1RD5 DRKETTTWISEGKLHASKILRFQKITENYLNVLYNCTVANEEAIDTKSFV

45 mIL-1RD1 DYQFVEHPSTKRKYTLITTLNISEVKSQFYRYPFICWKNTNIFESAHVQ

hIL-1RD1 DYYSVENPANKRRSTLITVLNISEIESRFYKHPFTCFAKNTHGIDAAYIQ

cIL-1RD1 QFYEEGMPHG..IAVSGTKFNISEVICLKDYAYKFFCHFIYDSQEFTSYIK

hIL-1RD8 GEIRLLKEHLG.EKEVELALIFDSWEADLAN.YTCHVENRNGRKHASVL

hFGR4 FPWQVLKTA DINSSEVEVLYLRNVSAED.AGEYTCLAGNSIGLSYQSAW

mIL-1RD3 VKQKV....IPPRYTVELACGFGATVFLWVLIWY

hIL-1RD6 LQLP.....APDFRAYLIGGLIALVAVAVSWYIYNIFKIDIVLWY

rIL-1RD6 LKRP.....APDFRAYLIGGLMAFLLLAVSILYIYNTFKVDIVLWY

mIL-iRD4 LRRK.....QPSKECPSHIAIYYIVAGCSLLLMFINVLVIVL

55 hIL-1RD4 LSRK.....NPSKEC

hIL-1RD2 VKEASS.....TFSWGIVLAPLSLAFLVLGGIWM

mIL-1RD2 VKEVSS.....TFSWSIALAPLSLIILWGAIW.

hIL-1RD10 LHKREL.....MYTVELAGGLGAILLLLVCLVTIYKCY

hIL-1RD5 LVRKADMADIP..GHVFTRGMIIAVLILVAWCLVTVCVIY

60 mIL-1RD5 LVRKEIPDIP...GHVFTGGVTVLVLASVAAVCIVILCVIY

mIL-1RD1 LIYP.....VPDFKNYLIGGFIILTATIVCCVCIY

hIL-1RD1 LIYP.....VTNFQKHMIGICVTLTVIIVCSVFIY

cIL-1RD1 LEHP.....VQNIRGYLIGGGISLIFLLFLILIVY

hIL-1RD8 LRKKDL.....IYKIELAGGLGAIFLLLVLLWIYKCY

hFGR4 LTVLP....E EDPTWTAAAP EARYTDIILY ASGSLALAVL LLLAGLY
Table 2 Alignment of the intracellular domains of various IL-lRs. hIL-1RD9 is SEQ ID NO: 8; mIL-1RD9 is.-GenBank is SEQ ID NO: 14;
hIL-1RD1 X16896; hIL-1RD6 is GenBank 065; mIL-1RD3 U49 is GenBank X85999;

huIL-1RD8 is GenBankY07519.
is SEQ
ID NO:
3; and mIL-1RD4 HuIL-1RD1SDGKTYDAYI LYPKTVGEG...STSDCDIFVFKVLPEVLEKQCGYKLFIY

HuIL-iRD6VDGKLYDAYV LYPKPHKES...QRHAVDALVLNILPEVLERQCGYKLFIF

MOIL-1RD3LDGKEYDIYV SYAR.........NVEEEEFVLLTLRGVLENEFGYKLCIF

HuIL-1RD8DDNKEYDAYL SYTKVDQDTLDCDNPEEEQFALEVLPDVLEKHYGY'KLFIP

HuIL-1RD5TDGKTYDAFV SYLKECRP....ENGEEHTFAVEILPRVLEKHFGYKLCIF

MOIL-1RD9.......... ................................

........
HuIL-1RD9.......... ...............................KYGYSLCLL

MOIL-1RD4NDGKLYDAYI IYPRVFRGS.AAGTHSVEYFVHHTLPDVLENKCGYKLCIY

HuIL-1RD1GRDDW.GED IVEVINENVKKSRRLIIILVRETSGFSWLGGSSEEQIAMY

HuIL-1RD6GRDEFP.GQA VANVIDENVKLCRRLIVIWPESLGFGLLKNLSEEQIAVY

MOIL-1RD3DRDSLPGGIV TDETLS.FIQKSRRLLWLSPNYVLQG.TQALLELKAGLE

HuIL-1RD8ERDLIPSG.T YMEDLTRYVEQSRRLIIVLTPDYILRR.GWSIFELESRLH

2 HuIL-1RD5ERDWPGGAV VDEIHS.LIEKSRRLIIVLSKSYMSN...EVRYELESGLH

MOIL-1RD9DRDVTP.GGV YADDIVSIIKKSRRGIFILSPSYLNG...PRVFELQAAVN

HuIL-1RD9ERDVAP.GGV YAEDIVSIIKRSRRGIFILSPNYVNG...PSIFELQAAVN

MOIL-1RD4GRDLLP.GQD AATWESSIQNSRRQVFVLAPHMMHSK..EFAYEQEIALH

3 HuIL-1RD1NALVQDGIKV VLLELEKIQ......DYEKMPESIKFIKQKHGAIRWSGDF
O

HuIL-1RD6SALIQDGMKV ILIELEKIE......DYTVMPESIQYIKQKHGAIRWHGDF

MOIL-1RD3NMASRGNINV ILVQYKAVK....DMKVKELKRAKTVLT....VIKWKGEK

HuIL-1RD8NMLVSGEIKV ILIECTELKGKVNCQEVESLKRSIKLLS....LIKWKGSK

HuIL-1RD5EALVERKIKI ILIEFTPVT......DFTFLPQSLKLLKSHR.VLKWKADK

3 MOIL-1RD9LALVDQTLKL ILIKFCSFQ......EPESLPYLVKKALRVLPTVTWKGLK

HuIL-1RD9LALDDQTLKL ILIKFCYFQ......EPESLPHLVKKALRVLPTVTWRGLK

MOIL-1RD4SALIQNNSKV ILIEMEPLG.EASRLQVGDLQDSLQHLVKIQGTIKWREDH

HuIL-1RD1 TQGPQSAKTR FWKNVRYHMP VQRRSPSSKH
40 HuIL-1RD6 TEQSQCMKTK FWKTVRYHMP PRRCRPFLRS

MOIL-1RD3 SKYPQ ...GRFWKQLQVAMP VKKSPRWSSN

HuIL-1RD8 SSKLN ...SKFWKHLVYEMP IKKKEMLPRC

HuIL-1RD5 SLSYN ...SRFWKNLLYLMP AKTVKPGRDE

MOIL-1RD9 SVHAS ...SRFWTQIRYHMP VKNSNRFMFN

45 HuIL-1RD9 SVPPN ...SRFWAKMRYHMP VKNSQGFTWN

Table 3 Alignment of primate IL-1RD8 and primate IL-1RD10.

R1D0 .......... .......... .......... .......... ..........

RD10 .......... .......... .......... .......... ..........

RD10 .......... .......... .......... .......... ..........

WO 99/19480 PCfNS98/20939 RD10 .............................. ....................

RD10 ......EFG..TSCELKYGGFVVRRTTELT VTAPLTDKPPKLLYPMESKL

RD8 SVIDVQLGKPLNIPCKAFFGFSGESGPMIY WMKGEKFIEELAG.HIREGE

RD10 TIQETQLGDSANLTCRAFFGYSGDVSPLIY WMKGEKFIEDLDENRVw7ESb O

RD8 KHLVYEMPIKKKEMLPRCHVLDSAEQGLFG ELQPIPSIAMTS.TSATLVS

RD8 SQADLP.EFHPS..DSMQIRHCCRGYKHEI PAT.TLPVPSLGNHHTYCNL

3 RD8 PLTLLNGQLPLNNTLKD..TQEFHRNSSLL PLSSKELSFTSDIW
O

Table 4 Alignment and comparison of primate and rodent IL-1RD9.
5 hIL-1RD9 MLCLGWIFLWLVAGERIKGFNISGCSTKKLLWTYSTRSEEEFVLFCDLPE
mIL-1RD9 MLCLGWVFLWFVAGEKTTGFNHSACATKKLLWTYSARGAENFVLFCDLQE
******,*** ****, *** * *,*********,* * *****ir*..k hIL-1RD9 PQKSHFCHRNRLSPKQVPEHLPFMGSN-DLSDVQWYQQPSNGDPLEDIRK
10 mIL-1RD9 LQEQKFSHASQLSPTQSPAHKPCSGSQKDLSDVQWYMQpRSGSPLEEISR
* ,*.* ,*** * * * * **, ******** ** * ***
hIL-1RD9 SYPHIIQDKCTLHFLTPGVNNSGSYICRPKMIKSPYDVACCVKMILEVKP
mIL-1RD9 NSPHMQSE-GMLHILAPQTNSIWSYICRPR-IRSPQDMACCIKTVLEVKP

15 **. . ** *,* * ******, *,** *,***.* ,*****~
hIL-1RD9 QTNASCEYSASHKQDLLLGSTGSISCPSLSCQSDAQSPAVTWYKNGKLLS
mIL-1RD9 QRNVSCGNTAQDEQVLLLGSTGSIHCPSLSCQSDVQSPEMTWYKDGRLLP
* * ** ,* * ********* ********* *** **** * **
hIL-1RD9 VERSNRIVVDEVYDYHQGTYVCDYTQSDTVSSWTVRAWQVRTIVGDTKL
mIL-1RD9 EHKKNPIEMADIYVFNQGLYVCDYTQSDNVSSWTVRAWKVRTIGKDINV
. * * . ..* ..** *********_**********,**** * , 2 5 hIL-1RD9 KPDILDPVEDTLEVELGKPLTISCKARFGFERVFNPVIKWYIKDSDLEWE
mIL-1RD9 KPEILDPITDTLDVELGKPLTLPCRVQFGFQRLSKPVIKWYVKESTQEWE
**,****, ***,********, *, ,***,*, ******.*,* ***
hIL-1RD9 VSVPEAKSIKSTLKDEIIERNIILEKVTQRDLRRKFVCFVQNSIGNTTQS
3 O mIL-1RD9 MSVFEEKRIQSTFKNEVIERTIFLREVTQRDLSRKFVCFAQNSIGNTTRT
,** * * *,** * *,***_* * ****** ****** ********
hIL-1RD9 VQLKEKRGVVLLYILLGTIGTLVAVLAASALLYRHWIEIVLLYRTYQSKD
mIL-1RD9 IRLRKKEEWFVYILLGTALMLVGVLVAAAFLYWYWIEWLLCRTYKNKD
3 5 ..*. * ** ,****** ** ** *_* ** ***,*** ***, **
hIL-1RD9 QTLGDKKDFDAFVSYAKWSSFPSEATSSLSEEHLALSLFPDVLENKYGYS
mIL-1RD9 ETLGDKKEFDAFVSYSNWSSPETDAVGSLSEEHLALNLFPEVLEDTYGYR
,******,*******, *** ,_* ********* ***,*** ***
hIL-1RD9 LCLLERDVAPGGVYAEDIVSIIKRSRRGIFILSPNYVNGPSIFELQAAVN
mIL-1RD9 LCLLDRDVTPGGVYADDIVSIIKKSRRGIFILSPSYLNGPRVFELQAAVN
****_***_******_******* ********** * *** ********
hIL-1RD9 LALDDQTLKLILiKFCYFQEPESLPHLVKKALRVLPTVTWRGLKSVPPNS
mIL-1RD9 LALVDQTLKLILIKFCSFQEPESLPYLVKKALRVLPTVTWKGLKSVHASS
*** ************ ******** ************** *****
hIL-1RD9 RFWAKMRYHMPVKNSQGFTWNQLRITSRIFQ-------WKGLSRTETTGR
5 0 mIL-1RD9 RFWTQIRYHMPVKNSNRFMFNGLRIFLKGFSPEKDLVTQKPLEGMPKSGN
***,..*********, * * *** , * * * ,*
hIL-1RD9 ----------SSQPKEW
mIL-1RD9 DHGAQNLLLYSDQKRC

Structural analysis of the primate IL-1RD10 sequence (SEQ ID NO: 18 and 20), in comparison with other IL-lRs, shows characteristic features exist, which are conserved with the IL-1RD10 embodiment described herein. For example, there are characteristic Ig domains, and" "
subdomains therein. The corresponding regions of the IL-1RD10 (SEQ ID N0: 18 and 20) are about: f2 to gly7; g2 from va110 to thr23; a3 from 1eu30 to met33; a3' from thr38 to g1n40; b3 from a1a48 to a1a54; c3 from pro64 to 1ys70; c3' from g1u72 to phe74; d3 from va183 to 1ys92;
e3 from g1n98 to va1106; and f3 from tyrll? to trp126.
Structural analysis of the rodent IL-1RD9 sequence (SEQ ID NO: 12, 14, and 16), in comparison with other IL-lRs, shows characteristic features exist (see Tables).
For example, there are characteristic Ig domains, and subdomains therein. The corresponding regions of the IL-1RD9 (SEQ ID NO: 12, 14, and 16) are about: Igl domain from g1y18 to pro127, with cys105 probably linked to cys52 (or possibly cys48); Ig2 domain from g1y128 to pro229, with cys153 probably linked to cys199; and the Ig3 domain from g1u230 to 1ys333, with cys251 probably linked to cys315; transmembrane segment from va1336 to tyr360; THD domain from g1y381 to va1539; conserved trp residues probably correspond to residues 64, 169, and 267. Alignment of the IL-1RD9 embodiments is shown in Table 4. There are characteristic beta strand sections, and alpha helical structures, as described above for IL-1RD10. The corresponding segments of the human IL-1RD9 sequence (SEQ ID NO: 6, 8, and 10) are roughly: ~iB from gly3 to va113; a2 from prol5 to 1ys28; ~3c from ser30 to ser46; a3 from i1e47 to g1n61; ~3D from 1ys64 to g1u75; a4 from g1u77 to 1eu87; (3E from va193 to 1eu98; and a5 from arg106 to va1117. The corresponding segments of the mouse IL-1RD9 sequence (SEQ ID N0: 12, 14, and 16) are roughly: oc3 to g1n10; ~iD from 1ys13 to g1u24; a4 from g1u26 to 1eu36; (3E from va42 to 1eu47; and oc5 from arg55 to va166.
As used herein, the terms IL-1 like receptor D8 (IL-1RD8), IL-1 like receptor D9 (IL-1RD9), or IL-1 like receptor D10 (IL-1RD10) shall be used to describe a polypeptide comprising a segment having or sharing the amino acid sequence shown in SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 18 and 20, or a substantial fragment thereof.
The invention also includes a polypeptide variation of the respective IL-1RD8, IL-1RD9, IL-1RD10 alleles whose sequences are provided, e.g., a mutein or soluble extracellular or intracellular construct. Typically, such agonists or antagonists will exhibit less than about 10~ sequence differences, and thus will often have between 1- and 11-fold substitutions, e.g., 2-, 3-, 5-, 7-fold, and others. It also encompasses allelic and other variants, e.g., natural polymorphic, of the polypeptide described. Typically, it will bind to its corresponding biological ligand, perhaps in a dimerized state with an alpha receptor subunit, with high affinity, e.g., at least about 100 nM, usually better than about 30 nM, preferably better than about 10 nM, and more preferably at better than about 3 nM. The term shall also be used herein to refer to related naturally occurring forms, e.g., alleles, polymorphic variants, and metabolic variants of the mammalian protein.
This invention also encompasses polypeptides having substantial amino acid sequence identity with the amino acid sequences shown in SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 18 and 20, preferably having segments of contiguous amino acid residues identical to segments of SEQ ID NO: 4, 10, or 20. It will include sequence variants with relatively few substitutions, e.g., typically less than about 25, ordinarily less than about 15, preferably less than about 3-5. Other embodiments include forms in association with an alpha subunit, e.g., an IL-1RD4, IL-1RD5, or IL-1RD6.
A substantial polypeptide "fragment", or "segment", is a stretch of amino acid residues of at least about 8 contiguous amino acids, generally at least 10 contiguous amino acids, more generally at least 12 contiguous amino acids, often at least 14 contiguous amino acids, more often at least 16 contiguous amino acids, typically at least 18 contiguous amino acids, more typically at least 20 contiguous amino acids, usually at least 22 contiguous amino acids, more usually at least 24 contiguous amino acids, preferably at least 26 contiguous amino acids, more preferably at least 28 contiguous amino acicT's,"and, in particularly preferred embodiments, at least about 30 or more contiguous amino acids, usually 40, 50, 70, 90, 110, etc. Sequences of segments of different polypeptides can be compared to one another over appropriate length stretches. In many cases, the matching will involve a plurality of distinct, e.g., nonoverlapping, segments of the specified length.
Typically, the plurality will be at least two, more usually at least three, and preferably 5, 7, or even more. While the length minima are provided, longer lengths, of various sizes, may be appropriate, e.g., one of length 7, and two of length 12. Similar features apply to segments of nucleic acid.
Amino acid sequence homology, or sequence identity, is determined by optimizing residue matches, if necessary, by introducing gaps as required. See, e.g., Needleham, et al. (1970) ~. Mol. Biol. 48:443-453;
Sankoff, et al. (1983) chapter one in Time Warps, String Edits, and Macromolecules: The Theory and Practice of Sequence Comparison. Addison-Wesley, Reading, MA; and software packages from IntelliGenetics, Mountain View, CA; and the University of Wisconsin Genetics Computer Group (GCG), Madison, WI; each of which is incorporated herein by reference. This changes when considering conservative substitutions as matches. Conservative substitutions typically include substitutions within the following groups: glycine, alanine; valine, isoleucine, leucine; aspartic acid, glutamic acid; asparagine, glutamine; serine, threonine; lysine, arginine; and phenylalanine, tyrosine. Homologous amino acid sequences are intended to include natural allelic and interspecies variations in the cytokine sequence. Typical homologous polypeptides will have from 50-100 homology (if gaps can ' be introduced), to 60-100$ homology (if conservative substitutions are included) with an amino acid sequence segment shown in SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 18 and 20. Homology measures will be at least about 70~, generally at least 76~, more generally at least 81~, often at least 85~, more often at least 88~, typically at least 90~, more typically at least 92~, usually at least 94~, more usually at least 95~, preferably at least 96~, and more preferably at least 97~, and in particularly preferred embodiments, at least 98~ or more. The degree of homology will vary with the length of the compared segments. Homologous polypeptides, such as the allelic variants, will share most biological activities with the embodiments described in SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 18 and 20.
As used herein, the term "biological activity" is used to describe, without limitation, effects on inflammatory responses, innate immunity, and/or morphogenic development by respective ligands. For example, these receptors should, like IL-1 receptors, mediate phosphatase or phosphorylase activities, which activities are easily measured by standard procedures.
See, e.g., Hardie, et al. (eds. 1995) The Protein Kinase FactBook vols. I and II, Academic Press, San Diego, CA;
Hanks, et al. (1991) Meth. Enzvmol. 200:38-62; Hunter, et al. (1992) Cell 70:375-388; Lewin (1990) Cell 61:743-752;
Pines, et al. (1991) Cold Snrina Harbor Sy~n~ Ouan~t Biol. 56:449-463; and Parker, et al. (2993) ur 363:736-738. Other activities include antigenic or immunogenic functions. The receptors exhibit biological activities much like regulatable enzymes, regulated by ligand binding. However, the enzyme turnover number is more close to an enzyme than a receptor complex.
Moreover, the numbers of occupied receptors necessary to induce such enzymatic activity is less than most receptor systems, and may number closer to dozens per cell, in contrast to most receptors which will trigger at numbers in the thousands per cell. The receptors, or portions.
thereof, may be useful as phosphate labeling enzymes to label general or specific substrates.

The terms ligand, agonist, antagonist, and analog of, e.g., an IL-1RD8, IL-1RD9, or IL-1RD10, include molecules that modulate the characteristic cellular responses to IL-1 ligand proteins, as well as molecules 5 possessing the more standard structural binding competition features of ligand-receptor interactions, e.g., where the receptor is a natural receptor or an antibody. The cellular responses likely are mediated through binding of various IL-1 ligands to cellular 10 receptors related to, but possibly distinct from, the type I or type II IL-1 receptors. See, e.g., Belvin and Anderson (1996) Ann. Rev. Cell Dev Biol 12:393-416;
Morisato and Anderson (1995) Ann. Rev Genetics 29:371-3991 and Hultmark (1994) Nature 367:116-117.
15 Also, a ligand is a molecule which serves either as a natural ligand to which said receptor, or an analog thereof, binds, or a molecule which is a functional analog of the natural ligand. The functional analog may be a ligand with structural modifications, or may be a 20 wholly unrelated molecule which has a molecular shape which interacts with the appropriate ligand binding determinants. The ligands may serve as agonists or antagonists, see, e.g., Goodman, et al. (eds. 1990) ~2odman & Gilman's: The Pharmacologvical Bases of Therat~eutics, Pergamon Press, New York.
Rational drug design may also be based upon structural studies of the molecular shapes of a receptor or antibody and other effectors or ligands. Effectors may be other proteins which mediate other functions in response to ligand binding, or other proteins which normally interact with the receptor. One means for determining which sites interact with specific other proteins is a physical structure determination, e.g., x-ray crystallography or 2 dimensional NMR techniques.
These will provide guidance as to which amino acid residues form molecular contact regions. For a detailed description of protein structural determination, see, e.g., Blundell and Johnson (1976) Protein ~rvstallocxra~y, Academic Press, New York, which is hereby incorporated herein by reference.
II. Activities The IL-1 receptor-like polypeptides will have a number of different biological activities, e.g., in phosphate metabolism, being added to or removed from specific substrates, typically proteins. Such will generally result in modulation of an inflammatory function, other innate immunity response, or a morphological effect. For example, a human IL-1RD9 gene coding sequence probably has about 60-80~ identity with the nucleotide coding sequence of mouse IL-1RD9. At the amino acid level, there is also likely to be reasonable identity.
The receptors will also exhibit immunogenic activity, e.g., in being capable of eliciting a selective immune response. Antiserum or antibodies resulting therefrom will exhibit both selectivity and affinity of binding. The polypeptides will also be antigenic, in binding antibodies raised thereto, in the native state, or in denatured.
The biological activities of the IL-lRDs will generally be related to addition or removal of phosphate moieties to substrates, typically in a specific manner, but occasionally in a non specific manner. Substrates may be identified, or conditions for enzymatic activity may be assayed by standard methods, e.g., as described in Hardie, et al. (eds. 1995) The Protein Kinase FactBook vols. I and II, Academic Press, San Diego, CA; Hanks, et al. (1991) Meth. Enzymol. 200:38-62; Hunter, et al.
(1992) Cell 70:375-388; Lewin (1990) Cell 61:743-752;
Pines, et al. (1991) Cold Snrina Harbor Symp Ouant Biol. 56:449-463; and Parker, et al. (1993) N ur 363:736-738.
III. Nucleic Acids This invention contemplates use of isolated nucleic acid or fragments, e.g., which encode these or closely related proteins, or fragments thereof, e.g., to encode a corresponding polypeptide, preferably one which is biologically active. In addition, this invention covers isolated or recombinant DNA which encodes such polypeptides or polypeptides having characteristic "
sequences of the respective IL-lRDs, individually or as a group. Typically, the nucleic acid is capable of hybridizing, under appropriate conditions, with a nucleic acid coding sequence segment shown in SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 17 or 19 but preferably not with a~
corresponding segment of other receptors. Said biologically active polypeptide can be a full length polypeptide, or fragment, and will typically have a segment of amino acid sequence highly homologous to one shown in SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 18 and 20. Further, this invention covers the use of isolated or recombinant nucleic acid, or fragments thereof, which encode polypeptides having fragments which are equivalent to the IL-1RD9 proteins. The isolated nucleic acids can have the respective regulatory sequences in the 5' and 3' flanks, e.g., promoters, enhancers, poly-A addition signals, and others from the natural gene.
An "isolated" nucleic acid is a nucleic acid, e.g., an RNA, DNA, or a mixed polymer, which is substantially pure, e.g., separated from other components which naturally accompany a native sequence, e.g., ribosomes, polymerases, and flanking genomic sequences from the originating species. The term embraces a nucleic acid sequence which has been removed from its naturally occurring environment, and includes recombinant or cloned DNA isolates, which are thereby distinguishable from naturally occurring compositions, and chemically synthesized analogs or analogs biologically synthesized by heterologous systems. A substantially pure molecule includes isolated forms of the molecule, either completely or substantially pure.
An isolated nucleic acid will generally be a homogeneous composition of molecules, but will, in some embodiments, contain heterogeneity, preferably minor.

This heterogeneity is typically found at the polymer ends or portions not critical to a desired biological function or activity.
A "recombinant" nucleic acid is typically defined either by its method of production or its structure'. In reference to its method of production, e.g., a product made by a process, the process is use of recombinant nucleic acid techniques, e.g., involving human intervention in the nucleotide sequence. Typically this intervention involves in vitro manipulation, although under certain circumstances it may involve more classical animal breeding techniques. Alternatively, it can be a nucleic acid made by generating a sequence comprising fusion of two fragments which are not naturally 25 contiguous to each other, but is meant to exclude products of nature, e.g., naturally occurring mutants as found in their natural state. Thus, for example, products made by transforming cells with an unnaturally occurring vector is encompassed, as are nucleic acids comprising sequence derived using any synthetic oligonucleotide process. Such a process is often done to replace a codon with a redundant codon encoding the same or a conservative amino acid, while typically introducing or removing a restriction enzyme sequence recognition site. Alternatively, the process is performed to join together nucleic acid segments of desired functions to generate a single genetic entity comprising a desired combination of functions not found in the commonly available natural forms, e.g., encoding a fusion protein.
Restriction enzyme recognition sites are often the target of such artificial manipulations, but other site specific targets, e.g., promoters, DNA replication sites, regulation sequences, control sequences, or other useful features may be incorporated by design. A similar concept is intended for a recombinant, e.g., fusion, polypeptide. This will include a dimeric repeat.
Specifically included are synthetic nucleic acids which, by genetic code redundancy, encode equivalent polypeptides to fragments of IL-1RD9 and fusions of sequences from various different related molecules, e.g., other IL-1 receptor family members.
A "fragment" in a nucleic acid context is a contiguous segment of at least about 17 contiguous nucleotides, generally at least 21 contiguous " "
nucleotides, more generally at least 25 contiguous nucleotides, ordinarily at least 30 contiguous nucleotides, more ordinarily at least 35 contiguous nucleotides, often at least 39 contiguous nucleotides, more often at least 45 contiguous nucleotides, typically at least 50 contiguous nucleotides, more typically at least 55 contiguous nucleotides, usually at least 60 contiguous nucleotides, more usually at least 66 contiguous nucleotides, preferably at least 72 contiguous nucleotides, more preferably at least 79 contiguous nucleotides, and in particularly preferred embodiments will be at least 85 or more contiguous nucleotides, e.g., 100, 120, 140, etc. Typically, fragments of different genetic sequences can be compared to one another over appropriate length stretches, particularly defined segments such as the domains described below.
A nucleic acid which codes for an IL-1RD8, IL-1RD9, or IL-1RD10 will be particularly useful to identify genes, mRNA, and cDNA species which code for itself or closely related proteins, as well as DNAs which code for polymorphic, allelic, or other genetic variants, e.g., from different individuals or related species. Preferred probes for such screens are those regions of the interleukin which are conserved between different polymorphic variants or which contain nucleotides which lack specificity, and will preferably be full length or nearly so. In other situations, polymorphic variant specific sequences will be more useful.
This invention further covers recombinant nucleic acid molecules and fragments having a nucleic acid sequence identical to or highly homologous to the isolated DNA set forth herein. In particular, the sequences will often be operably linked to DNA segments which control transcription, translation, and DNA

replication. These additional segments typically assist in expression of the desired nucleic acid segment.
Homologous, or highly identical, nucleic acid sequences, when compared to one another, e.g., IL-1RD9 5 sequences, exhibit significant similarity. The standards for homology in nucleic acids are either measures for homology generally used in the art by sequence comparison or based upon hybridization conditions. Comparative hybridization conditions are described in greater detail 10 below. ' Substantial identity in the nucleic acid sequence comparison context means either that the segments, or their complementary strands, when compared, are identical when optimally aligned, with appropriate nucleotide 15 insertions or deletions, in at least about 60~ of the nucleotides, generally at least 66$, ordinarily at least 71~, often at least 76~, more often at least 80~, usually at least 84~, more usually at least 88~, typically at least 91~, more typically at least about 93~, preferably 20 at least about 95~, more preferably at least about 96 to 98~ or more, and in particular embodiments, as high at about 99~ or more of the nucleotides, including, e.g., segments encoding structural domains such as the segments described below. Alternatively, substantial identity 25 will exist when the segments will hybridize under selective hybridization conditions, to a strand or its complement, typically using a sequence derived from SEQ
ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 17 or 19. Typically, selective hybridization will occur when there is at least about 55~ homology over a stretch of at least about 14 nucleotides, more typically at least about 65~, preferably at least about 75~, and more preferably at least about 90~. See, Kanehisa (1984) Nuc. Acids Res.
12:203-213, which is incorporated herein by reference.
The length of homology comparison, as described, may be over longer stretches, and in certain embodiments will be over a stretch of at least about 17 nucleotides, .' generally at least about 20 nucleotides, ordinarily at least about 24 nucleotides; usually at least about 28 nucleotides, typically at least about 32 nucleotides, more typically at least about 40 nucleotides, preferably at least about 50 nucleotides, and more preferably at least about 75 to 100 or more nucleotides.
Stringent conditions, in referring to homology'in the hybridization context, will be stringent combined conditions of salt, temperature, organic solvents, and other parameters typically controlled in hybridization reactions. Stringent temperature conditions will usually include temperatures in excess of about 30' C, more usually in excess of about 37' C, typically in excess of about 45'. C, more typically in excess of about 55' C, preferably in excess of about 65' C, and more preferably in excess of about 70' C. Stringent salt conditions will ordinarily be less than about 500 mM, usually less than about 400 mM, more usually less than about 300 mM, typically less than about 200 mM, preferably less than about 100 mM, and more preferably less than about 80 mM, even down to less than about 20 mM. However, the combination of parameters is much more important than the measure of any single parameter. See, e.g., Wetmur and Davidson (1968) J. Mol. Biol. 31:349-370, which is hereby incorporated herein by reference. . The signal should be at least 2X over background, generally at least 5-lOX
over background, and preferably even more.
For sequence comparison, typically one sequence acts as a reference sequence, to which test sequences are compared. When using a sequence comparison algorithm, test and reference sequences are input into a computer, subsequent coordinates are designated, if necessary, and sequence algorithm program parameters are designated.
The sequence comparison algorithm then calculates the percent sequence identity for the test sequences) relative to the reference sequence, based on the designated program parameters.
Optical alignment of sequences for comparison can be conducted, e.g., by the local homology algorithm of Smith and Waterman (1981) Adv. At~pl. Math. 2:482, by the homology alignment algorithm of Needleman and Wunsch (197,0) J. Mol. Biol. 48:443, by the search for similarity method of Pearson and Lipman (1988) Proc. Nat~l Acad Sci. USA 85:2444, by computerized implementations of these algorithms (GAP, BESTFIT, FASTA, and TFASTA in the Wisconsin Genetics Software Package, Genetics Compu'~er Group, 575 Science Dr., Madison, WI), or by visual inspection (see generally Ausubel et al., supra).
One example of a useful algorithm is PILEUP. PILEUP
creates a multiple sequence alignment from a group of related sequences using progressive, pairwise alignments to show relationship and percent sequence identity. It also plots a tree or dendrogram showing the clustering relationships used to create the alignment. PILEUP uses a simplification of the progressive alignment method of Feng and Doolittle (1987) J. Mol. Evol. 35:351-360. The method used is similar to the method described by Higgins and Sharp (1989) CABIOS 5:151-153. The program can align up to 300 sequences, each of a maximum length of 5,000 nucleotides or amino acids. The multiple alignment procedure begins with the pairsaise alignment of the two most similar sequences, producing a cluster of two aligned sequences. This cluster is then aligned to the next most related sequence or cluster of aligned sequences. Two clusters of sequences are aligned by a simple extension of the pairwise alignment of two individual sequences. The final alignment is achieved by a series of progressive, pairwise alignments. The program is run by designating specific sequences and their amino acid or nucleotide coordinates for regions of sequence comparison and by designating the program parameters. For example, a reference sequence can be compared to other test sequences to determine the percent sequence identity relationship using the following parameters: default gap weight (3.00), default gap length weight (0.10), and weighted end gaps.
Another example of algorithm that is suitable for determining percent sequence identity and sequence similarity is the BLAST algorithm, which is described Altschul, et al. (1990) U. Mol. Biol. 215:403-410.

Software for performing BLAST analyses is publicly available through the National Center for Biotechnology Information (http:www.ncbi.nlm.nih.gov/). This algorithm involves first identifying high scoring sequence pairs (HSPs) by identifying short words of length W in "the query sequence, which either match or satisfy some positive-valued threshold score T when aligned with a word of the same length in a database sequence. T is referred to as the neighborhood word score threshold (Altschul, et al., supra). These initial neighborhood word hits act as seeds for initiating searches to find longer HSPs containing them. The word hits are then extended in both directions along each sequence for as far as the cumulative alignment score can be increased.
Extension of the word hits in each direction are halted when: the cumulative alignment score falls off by the quantity X from its maximum achieved value; the cumulative score goes to zero or below, due to the accumulation of one or more negative-scoring residue alignments; or the end of either sequence is reached.
The BLAST algorithm parameters W, T, and X determine the sensitivity and speed of the alignment. The BLAST
program uses as defaults a wordlength (W) of 11, the BLOSUM62 scoring matrix (see Henikoff and Henikoff (1989) Proc Nat'1 Acad Sci USA 89:10915) alignments (B) of 50, expectation (E) of 10, M=5, N=4, and a comparison of both strands.
In addition to calculating percent sequence identity, the BLAST algorithm also performs a statistical analysis of the similarity between two sequences (see, e.g., Karlin and Altschul (1993) Proc. Nat'1 Acad ci-~ 90:5873-5787). One measure of similarity provided by the BLAST algorithm is the smallest sum probability (P(N)), which provides an indication of the probability by which a match between two nucleotide or amino acid sequences would occur by chance. For example, a nucleic acid is considered similar to a reference sequence if the smallest sum probability in a comparison of the test nucleic acid to the reference nucleic acid is less than about 0.1, more preferably less than about 0.01, and most preferably less than about 0.001.
A further indication that two nucleic acid sequences of polypeptides are substantially identical is that the polypeptide encoded by the first nucleic acid is "
immunologically cross reactive with the polypeptide encoded by the second nucleic acid, as described below.
Thus, a polypeptide is typically substantially identical to a second polypeptide, e.g., where the two peptides differ only by conservative substitutions. Another indication that two nucleic acid sequences are substantially identical is that the two molecules hybridize to each other under stringent conditions, as described below.
The isolated DNA can be readily modified by nucleotide substitutions, nucleotide deletions, nucleotide insertions, and inversions of nucleotide stretches. These modifications result in novel DNA
sequences which encode this polypeptide or its derivatives. These modified sequences can be used to produce mutant proteins (muteins) or to enhance the expression of variant species. Enhanced expression may involve gene amplification, increased transcription, increased translation, and other mechanisms. Such mutant IL-1RD9-like derivatives include predetermined or site-specific mutations of the polypeptide or its fragments, including silent mutations using genetic code degeneracy. "Mutant IL-1RD9" as used herein encompasses a polypeptide otherwise falling within the homology definition of the IL-1R9 as set forth above, but having an amino acid sequence which differs from that of other IL-1RD-like polypeptides as found in nature, whether by way of deletion, substitution, or insertion. In particular, "site specific mutant IL-1RD9" encompasses a polypeptide having substantial homology with a polypeptide of SEQ ID NO: 6, 8, 10, 12, 14 or 14, and typically shares most of the biological activities or effects of the forms disclosed herein.

Although site specific mutation sites are predetermined, mutants need not be site specific.
Mammalian IL-1RD9 mutagenesis can be achieved by making amino acid insertions or deletions in the gene, coupled 5 with expression. Substitutions, deletions, insertions, or many combinations may be generated to arrive at a final construct. Insertions include amino- or carboxy-terminal fusions. Random mutagenesis can be conducted at a target codon and the expressed mammalian IL-1RD9 10 mutants can then be screened for the desired activity, providing some aspect of a structure-activity relationship. Methods for making substitution mutations at predetermined sites in DNA having a known sequence are well known in the art, e.g., by M13 primer mutagenesis.
15 See also Sambrook, et al. (1989) and Ausubel, et al.
(1987 and periodic Supplements).
The mutations in the DNA normally should not place coding sequences out of reading frames and preferably will not create complementary regions that could 20 hybridize to produce secondary mRNA structure such as loops or hairpins.
The phosphoramidite method described by Beaucage and Carruthers (1981) Tetra. Letts. 22:1859-1862, will produce suitable synthetic DNA fragments. A double 25 stranded fragment will often be obtained either by synthesizing the complementary strand and annealing the strand together under appropriate conditions or by adding the complementary strand using DNA polymerase with an appropriate primer sequence.

30 Polymerase chain reaction (PCR) techniques can often be applied in mutagenesis. Alternatively, mutagenesis primers are commonly used methods for generating defined mutations at predetermined sites. See, e.g., Innis, et al. (eds. 1990) PCR Protocol s A Guide to Methods and A~~lications Academic Press, San Diego, CA; and Dieffenbach and Dveksler (1995; eds.) PCB Primer: A
~boratorv Manual Cold Spring Harbor Press, CSH, NY.
Appropriate primers of length, e.g., 15, 20, 25, or longer can be made using sequence provided.

IV. Proteins, Peptides As described above, the present invention encompasses primate IL-1RD8, primate or rodent IL-1RD9, and primate IL-1RD10,e.g., whose sequences are disclosed e.g., in Tables 1-3, and described herein. Descriptions of features of IL-1RD9 are applicable in most cases, with appropriate modifications, also to IL-1RD8 and/or to IL-1RD10. Allelic and other variants are also contemplated, including, e.g., fusion proteins combining portions of such sequences with others, including epitope tags and functional domains. Particularly interesting constructs will be intact extracellular or intracellular domains.
The present invention also provides recombinant polypeptides, e.g., heterologous fusion proteins using segments from these rodent proteins. A heterologous fusion protein is a fusion of proteins or segments which are naturally not normally fused in the same manner.
Thus, the fusion product of, e.g., an IL-1RD9 with another IL-1 receptor is a continuous protein molecule having sequences fused in a typical polypeptide linkage, typically made as a single translation product and exhibiting properties, e.g., sequence or antigenicity, derived from each source peptide. A similar concept applies to heterologous nucleic acid sequences.
In addition, new constructs may be made from combining similar functional or structural domains from other related proteins, e.g., IL-1 receptors or Toll-like receptors, including species variants. For example, ligand-binding or other segments may be "swapped" between different new fusion polypeptides or fragments. See, e.g., Cunningham, et al. (1989) Science 243:1330-1336;
and O'Dowd, et al. (1988) J. Biol. Chem. 263:15985-15992, each of which is incorporated herein by reference. Thus, new chimeric polypeptides exhibiting new combinations of specificities will result from the functional linkage of receptor-binding specificities. For example, the ligand binding domains from other related receptor molecules may be added or substituted for other domains of this or related proteins. The resulting protein will often have hybrid function and properties. For example, a fusion protein may include a targeting domain which may serve to provide sequestering of the fusion protein to a particular subcellular organelle. "
Candidate fusion partners and sequences can be selected from various sequence data bases, e.g., GenBank, c/o NCBI, and BCG, University of Wisconsin Biotechnology Computing Group, Madison, WI, which are each incorporated herein by reference.
The present invention particularly provides muteins which bind IL-1-like ligands, and/or which are affected in signal transduction. Structural alignment of human IL-1RD9 with other members of the IL-1R family show conserved features/residues. See Tables 1-4. Alignment of the human IL-1RD9 sequence with other members of the IL-1R family indicates various structural and functionally shared features. See also, Bazan, et al.
(1996) ature 379:591; Lodi, et al. (1994) Science 263:1762-1766; Sayle and Milner-White (1995) TIB 20:374-376; and Gronenberg, et al. (1991) Protein Enaineerina 4:263-269.
The IL-loc and IL-1(3 ligands bind an IL-1 receptor type I (IL-1RD1) as the primary receptor and this complex then forms a high affinity receptor complex with the IL-1 receptor type III (IL-1RD3). Such receptor subunits are probably shared with the receptors for the new IL-1 ligand family members. See, e.g., USSN 60/044,165 and USSN 60/055,111. It is likely that the IL-1'y ligand signals through a receptor comprising the association of IL-1RD9 (alpha component) with IL-1RD5 (beta component).
The IL-1S and IL-1~ ligands each probably signal through a receptor comprising the association of one of IL-1RD4, IL-1RD6, or IL-1RD9 (alpha components) with one of IL-1RD3, IL-1RD5, IL-1RD7, IL-1RD8, or IL-1RD10 (beta components).
Similar variations in other species counterparts of IL-1R sequences, e.g., receptors D1-D6, D8, D9, or D10, in the corresponding regions, should provide similar interactions with ligand or substrate. Substitutions with either rodent or primate, e.g., mouse sequences or human sequences, are particularly preferred. Conversely, conservative substitutions away from the ligand binding interaction regions will probably preserve most signaling activities; and conservative substitutions away from the intracellular domains will probably preserve most ligand binding properties.
"Derivatives" of the primate or mouse IL-1RD9 include amino acid sequence mutants, glycosylation variants, metabolic derivatives and covalent or aggregative conjugates with other chemical moieties.
Covalent derivatives can be prepared by linkage of functionalities to groups which are found in the IL-1RD9 amino acid side chains or at the N- or C- termini, e.g., by means which are well known in the art. These derivatives can include, without limitation, aliphatic esters or amides of the carboxyl terminus, or of residues containing carboxyl side chains, O-acyl derivatives of hydroxyl group-containing residues, and N-acyl derivatives of the amino ternninal amino acid or amino-group containing residues, e.g., lysine or arginine. Acyl groups are selected from the group of alkyl-moieties including C3 to C18 normal alkyl, thereby forming alkanoyl aroyl species.
In particular, glycosylation alterations are included, e.g., made by modifying the glycosylation patterns of a polypeptide during its synthesis and processing, or in further processing steps. Particularly preferred means for accomplishing this are by exposing the polypeptide to glycosylating enzymes derived from cells which normally provide such processing, e.g., mammalian glycosylation enzymes. Deglycosylation enzymes are also contemplated. Also embraced are versions of the same primary amino acid sequence which have other minor modifications, including phosphorylated amino acid residues, e.g., phosphotyrosine, phosphoserine, or phosphothreonine.

A major group of derivatives are covalent conjugates of the receptors or fragments thereof with other polypeptides. These derivatives can be synthesized in recombinant culture such as N- or C-terminal fusions or by the use of agents known in the art for their " "
usefulness in cross-linking proteins through reactive side groups. Preferred derivatization sites with cross-linking agents are at free amino groups, carbohydrate moieties, and cysteine residues.
Fusion polypeptides between the receptors and other homologous or heterologous proteins are also provided.
Homologous polypeptides may be fusions between different receptors, resulting in, for instance, a hybrid protein exhibiting binding specificity for multiple different IL-1 ligands, or a receptor which may have broadened or weakened specificity of substrate effect. Likewise, heterologous fusions may be constructed which would exhibit a combination of properties or activities of the derivative proteins. Typical examples are fusions of a reporter polypeptide, e.g., luciferase, with a segment or domain of a receptor, e.g., a ligand-binding segment, so that the presence or location of a desired ligand may be easily determined. See, e.g., Dull, et al., U.S. Patent No. 4,859,609, which is hereby incorporated herein by reference. Other gene fusion partners include glutathione-S-transferase (GST), bacterial i~-galactosidase, trpE, Protein A, Q-lactamase, alpha amylase, alcohol dehydrogenase, and yeast alpha mating factor. See, e.g., Godowski, et al. (1988) Science 241:812-816.
The phosphoramidite method described by Beaucage and Carruthers (1981) Tetra. Letts. 22:1859-1862, will produce suitable synthetic DNA fragments. A double stranded fragment will often be obtained either by synthesizing the complementary strand and annealing the strand together under appropriate conditions or by adding the complementary strand using DNA polymerase with an appropriate primer sequence.

Such polypeptides may also have amino acid residues which have been chemically modified by phosphorylation, sulfonation, biotinylation, or the addition or removal of other moieties, particularly those which have molecular 5 shapes similar to phosphate groups. In some embo3i'Qhents, the modifications will be useful labeling reagents, or serve as purification targets, e.g., affinity ligands.
Fusion proteins will typically be made by either recombinant nucleic acid methods or by synthetic 10 polypeptide methods. Techniques for nucleic acid manipulation and expression are described generally, e.g., in Sambrook, et al. (1989) Molecular Cloning: A
Laboratory Manual (2d ed.), Vols. 1-3, Cold Spring Harbor Laboratory, and Ausubel, et al. (eds. 1987 and periodic 15 supplements) Current Protocols in Molecular Bioloav, Greene/Wiley, New York, which are each incorporated herein by reference. Techniques for synthesis of polypeptides are described, e.g., in Merrifield (1963) Amer. Chem. Soc. 85:2149-2156; Merrifield (1986) science 20 232: 341-347; and Atherton, et al. (1989) Solid Phase Peptide Synthesis: A Practic~,,'~ Approach, IRL Press, Oxford; each of which is incorporated herein by reference. See also Dawson, et al. (1994) Science 266:776-779 fox methods to make larger polypeptides.
25 This invention also contemplates the use of derivatives of an IL-1RD8, IL-1RD9, or IL-1RD10 other than variations in amino acid sequence or glycosylation.
Such derivatives may involve covalent or aggregative association with chemical moieties. These derivatives 30 generally fall into three classes: (1) salts, (2) side chain and terminal residue covalent modifications, and (3) adsorption complexes, for example with cell membranes. Such covalent or aggregative derivatives are useful as immunogens, as reagents in immunoassays, or in 35 purification methods such as for affinity purification of a receptor or other binding molecule, e.g., an antibody.
For example, an IL-1 ligand can be immobilized by covalent bonding to a solid support such as cyanogen bromide-activated Sepharose, by methods which are well known in the art, or adsorbed onto polyolefin surfaces, with or without glutaraldehyde cross-linking, for use in the assay or purification of an IL-1 receptor, antibodies, or other similar molecules. The ligand can also be labeled with a detectable group, e.g., radioiodinated by the chloramine T procedure, covalently bound to rare earth chelates, or conjugated to another fluorescent moiety for use in diagnostic assays.
An IL-1RD8, IL-1RD9, or IL-1RD10 of this invention can be used as an immunogen for the production of antisera or antibodies specific, e.g., capable of distinguishing between other IL-1 receptor family members, for the IL-1RD8, IL-1RD9, or IL-1RD10 or various fragments thereof. The purified IL-1RD8, IL-1RD9, or IL-1RD10 can be used to screen monoclonal antibodies or antigen-binding fragments prepared by immunization with various forms of impure preparations containing the protein. In particular, the term "antibodies" also encompasses antigen binding fragments of natural antibodies, e.g., Fab, Fab2, Fv, etc. The purified IL-1RD9 can also be used as a reagent to detect antibodies generated in response to the presence of elevated levels of expression, or immunological disorders which lead to antibody production to the endogenous receptor.
Additionally, IL-1RD8, IL-1RD9, or IL-1RD10 fragments may also serve as immunogens to produce the antibodies of the present invention, as described immediately below. For example, this invention contemplates antibodies having binding affinity to or being raised against the amino acid sequences shown, e.g., in SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 18 and 20, fragments thereof, or various homologous peptides. In particular, this invention contemplates antibodies having binding affinity to, or having been raised against, specific fragments which are predicted to be, or actually are, exposed at the exterior polypeptide surface of the native IL-1RD8, IL-1RD9, or IL-1RD10. Various preparations of desired selectivity~in binding can be prepared by appropriate cross absorptions, ' etc.

The blocking of physiological response to the receptor ligands may result from the inhibition of binding of the ligand to the receptor, likely through competitive inhibition. Thus, in vitro assays of the present invention will often use antibodies or an~i~en binding segments of these antibodies, or fragments attached to solid phase substrates. These assays will also allow for the diagnostic determination of the effects of either ligand binding region mutations and modifications, or other mutations and modifications, e.g., which affect signaling or enzymatic function.
This invention also contemplates the use of competitive drug screening assays, e.g., where neutralizing antibodies to the receptor or fragments compete with a test compound for binding to a ligand or other antibody. In this manner, the neutralizing antibodies or fragments can be used to detect the presence of a polypeptide which shares one or more binding sites to a receptor and can also be used to occupy binding sites on a receptor that might otherwise bind a ligand.
V. Making Nucleic Acids and Protein DNA which encodes the polypeptides or fragments thereof can be obtained by chemical synthesis, screening cDNA libraries, or by screening genomic libraries prepared from a wide variety of cell lines or tissue samples. Natural sequences can be isolated using standard methods and the sequences provided herein, e.g., in Tables 1-3. Other species counterparts can be identified by hybridization techniques, or by various PCR
techniques, combined with or by searching in sequence databases, e.g., GenBank.
This DNA can be expressed in a wide variety of host cells for the synthesis of a full-length receptor or fragments which can in turn, e.g., be used to generate polyclonal or monoclonal antibodies; for binding studies;
for construction and expression of modified ligand binding or kinase/phosphatase domains; and for structure/function studies. Variants or fragments can be expressed in host cells that are transformed or transfected with appropriate expression vectors. These molecules can be substantially free of protein or cellular contaminants, other than those derived from the recombinant host, and therefore are particularly useful in pharmaceutical compositions when combined with a pharmaceutically acceptable carrier and/or diluent. The protein, or portions thereof, may be expressed as fusions with other proteins.
Expression vectors are typically self-replicating DNA or RNA constructs containing the desired receptor gene or its fragments, usually operably linked to suitable genetic control elements that are recognized in a suitable host cell. These control elements are capable of effecting expression within a suitable host. The specific type of control elements necessary to effect expression will depend upon the eventual host cell used.
Generally, the genetic control elements can include a prokaryotic promoter system or a eukaryotic promoter expression control system, and typically include a transcriptional promoter, an optional operator to control the onset of transcription, transcription enhancers to elevate the level of mRNA expression, a sequence that encodes a suitable ribosome binding site, and sequences that terminate transcription and translation. Expression vectors also usually contain an origin of replication that allows the vector to replicate independently of the host cell.
The vectors of this invention include those which contain DNA which encodes a protein, as described, or a fragment thereof encoding a biologically active equivalent polypeptide. The DNA can be under the control of a viral promoter and can encode a selection marker.
This invention further contemplates use of such expression vectors which are capable of expressing eukaryotic cDNA coding for such a polypeptide in a prokaryotic or eukaryotic host, where the vector is compatible with the host and where the eukaryotic cDNA

coding for the receptor is inserted into the vector such that growth of the host containing the vector expresses the cDNA in question. Usually, expression vectors are designed for stable replication in their host cells or for amplification to greatly increase the total number of copies of the desirable gene per cell. It is not always necessary to require that an expression vector replicate in a host cell, e.g., it is possible to effect transient expression of the polypeptide or its fragments in various hosts using vectors that do not contain a replication origin that is recognized by the host cell. It is also possible to use vectors that cause integration of the polypeptide encoding portion or its fragments into the host DNA by recombination.
Vectors, as used herein, comprise plasmids, viruses, bacteriophage, integratable DNA fragments, and other vehicles which enable the integration of DNA fragments into the genome of the host. Expression vectors are specialized vectors which contain genetic control elements that effect expression of operably linked genes.
Plasmids are the most commonly used form of vector but all other forms of vectors which serve an equivalent function and which are, or become, known in the art are suitable for use herein. See, e.g., Pouwels, et al.
(1985 and Supplements) Clonincr Vectors: A Laboratory anual, Elsevier, N.Y., and Rodriquez, et al. (eds.) Vectors: A Survey of Molecular Clonina Vectors and Their Uses, Buttersworth, Boston, 1988, which are incorporated herein by reference.
Transformed cells are cells, preferably mammalian, that have been transformed or transfected with receptor vectors constructed using recombinant DNA techniques.
Transformed host cells usually express the desired polypeptide or its fragments, but for purposes of cloning, amplifying, and manipulating its DNA, do not need to express the subject protein. This invention further contemplates culturing transformed cells in a nutrient medium, thus permitting the receptor to accumulate in the cell membrane. The polypeptide can be recovered, either from the culture or, in certain instances, from the culture medium.
For purposes of this invention, nucleic sequences are operably linked when they are functionally related to 5 each other. For example, DNA for a presequence or secretory leader is operably linked to a polypeptide if it is expressed as a preprotein or participates in directing the polypeptide to the cell membrane or in secretion of the polypeptide. A promoter is operably 10 linked to a coding sequence if it controls the transcription of the polypeptide; a ribosome binding site is operably linked to a coding sequence if it is positioned to permit translation. Usually, operably linked means contiguous and in reading frame, however, 15 certain genetic elements such as repressor genes are not contiguously linked but still bind to operator sequences that in turn control expression.
Suitable host cells include prokaryotes, lower eukaryotes, and higher eukaryotes. Prokaryotes include 20 both gram negative and gram positive organisms, e.g., coli and B. subtilis. Lower eukaryotes include yeasts, e.g., ~,. cerevisiae and Pichia, and species of the genus Di~tyostelium. Higher eukaryotes include established tissue culture cell lines from animal cells, both of 25 non-mammalian origin, e.g., insect cells, and birds, and of mammalian origin, e.g., human, primates, and rodents.
Prokaryotic host-vector systems include a wide variety of vectors for many different species. As used herein, E. coli and its vectors will be used generically 30 to include equivalent vectors used in other prokaryotes.
A representative vector for amplifying DNA is pBR322 or many of its derivatives. Vectors that can be used to express the receptor or its fragments include, but are not limited to, such vectors as those containing the lac 35 promoter (pUC-series); trp promoter (pBR322-trp); Ipp promoter (the pIN-series); lambda-pP or pR promoters (pOTS); or hybrid promoters such as ptac (pDR540). See Brosius, et al. (1988) "Expression Vectors Employing Lambda-, trp-, lac-, and Ipp-derived Promoters", in Vectors: A Survey of Molecular Cloninct Vectors and Their Uses, (eds. Rodriguez and Denhardt), Buttersworth, Boston, Chapter ~10, pp. 205-236, which is incorporated herein by reference.
Lower eukaryotes, e.g., yeasts and Dictvoste~ium, may be transformed with IL-1RD9 sequence containing vectors. For purposes of this invention, the most common lower eukaryotic host is the baker's yeast, Saccharomyces cerevisiae. It will be used to generically represent lower eukaryotes although a number of other strains and species are also available. Yeast vectors typically consist of a replication origin (unless of the integrating type), a selection gene, a promoter, DNA
encoding the receptor or its fragments, and sequences for translation termination, polyadenylation, and transcription termination. Suitable expression vectors for yeast include such constitutive promoters as 3-phosphoglycerate kinase and various other glycolytic enzyme gene promoters or such inducible promoters as the alcohol dehydrogenase 2 promoter or metallothionine promoter. Suitable vectors include derivatives of the following types: self-replicating low copy number (such as the YRp-series), self-replicating high copy number (such as the YEp-series); integrating types (such as the YIp-series), or mini-chromosomes (such as the YCp-series).
Higher eukaryotic tissue culture cells are normally the preferred host cells for expression of the functionally active interleukin protein. In principle, many higher eukaryotic tissue culture cell lines are workable, e.g., insect baculovirus expression systems, whether from an invertebrate or vertebrate source.
However, mammalian cells are preferred. Transformation or transfection and propagation of such cells has become a routine procedure. Examples of useful cell lines include HeLa cells, Chinese hamster ovary (CHO) cell lines, baby rat kidney (BRK) cell lines, insect cell lines, bird cell lines, and monkey (COS) cell lines.
Expression vectors for such cell lines usually include an WO 99/19480 PCT/US98l20939 origin of replication, a promoter, a translation initiation site, RNA splice sites (if genomic DNA is used), a polyadenylation site, and a transcription termination site. These vectors also usually contain a selection gene or amplification gene. Suitable expression vectors may be plasmids, viruses, or retroviruses carrying promoters derived, e.g., from such sources as from adenovirus, SV40, parvoviruses, vaccinia virus, or cytomegalovirus. Representative examples of suitable expression vectors include pCDNAI; pCD, see Okayama, et al. (1985) Mol. Cell Biol. 5:1136-1142;
pMClneo PolyA, see Thomas, et al. (1987) Cell 51:503-512;
and a baculovirus vector such as pAC 373 or pAC 610.
For secreted proteins, an open reading frame usually encodes a polypeptide that consists of a mature or secreted product covalently linked at its N-terminus to a signal peptide. The signal peptide is cleaved prior to secretion of the mature, or active, polypeptide. The cleavage site can be predicted with a high degree of accuracy from empirical rules, e.g., von-Heijne (1986) Nucleic Acids Research 14:4683-4690 and Nielsen, et al.
(1997) Protein Enct. 10:1-12, and the precise amino acid composition of the signal peptide often does not appear to be critical to its function, e.g., Randall, et al.
(1989) Science 243:1156-1159; Kaiser, et al. (1987) science 235:312-317.
It will often be desired to express these polypeptides in a system which provides a specific or defined glycosylation pattern. In this case, the usual pattern will be that provided naturally by the expression system. However, the pattern will be modifiable by exposing the polypeptide, e.g., an unglycosylated form, to appropriate glycosylating proteins introduced into a heterologous expression system. For example, the receptor gene may be co-transformed with one or more genes encoding mammalian or other glycosylating enzymes.
Using this approach, certain mammalian glycosylation patterns will be achievable in prokaryote or other cells.

43 _ The source of IL-1RD8, IL-1RD9, or IL-1RD10 can be a eukaryotic or prokaryotic host expressing recombinant IL-1RD8, IL-1RD9, or IL-1RD10 such as is described above.
The source can also be a cell line such as mouse Swiss 3T3 fibroblasts, but other mammalian cell lines are~'also contemplated by this invention, with the preferred cell line being from the human species.
Now that the sequences are known, the primate IL-lRs, fragments, or derivatives thereof can be prepared by conventional processes for synthesizing peptides. These include processes such as are described in Stewart and Young (1984) Solid Phase Peptide Synthesis, Pierce Chemical Co., Rockford, IL; Bodanszky and Bodanszky (1984) The Practice of Peptide Synthesis, Springer-Verlag, New York; and Bodanszky (1984) ~e Principles of P~,ptide Synthesis, Springer-Verlag, New York; all of each which are incorporated herein by reference. For example, an azide process, an acid chloride process, an acid anhydride process, a mixed anhydride process, an active ester process (e. g., p-nitrophenyl ester, N-hydroxysuccinimide ester, or cyanomethyl ester), a carbodiimidazole process, an oxidative-reductive process, or a dicyclohexylcarbodiimide (DCCD)/additive process can be used. Solid phase and solution phase syntheses are both applicable to the foregoing processes. Similar techniques can be used with partial IL-1RD9 sequences.
The IL-1RD8, IL-1RD9, or IL-1RD10 proteins, polypeptides, fragments, or derivatives are suitably prepared in accordance with the above processes as typically employed in peptide synthesis, generally either by a so-called stepwise process which comprises condensing an amino acid to the terminal amino acid, one by one in sequence, or by coupling peptide fragments to the terminal amino acid. Amino groups that are not being used in the coupling reaction typically must be protected to prevent coupling at an incorrect location.
If a solid phase synthesis is adopted, the C-terminal amino acid is bound to an insoluble carrier or support through its carboxyl group. The insoluble carrier is not particularly limited as long as it has a binding capability to a reactive carboxyl group.
Examples of such insoluble carriers include halomethyl resins, such as chloromethyl resin or bromomethyh resin, hydroxymethyl resins, phenol resins, tert-alkyloxycarbonylhydrazidated resins, and the like.
An amino group-protected amino acid is bound in sequence through condensation of its activated carboxyl group and the reactive amino group of the previously, formed peptide or chain, to synthesize the peptide step by step. After synthesizing the complete sequence, the peptide is split off from the insoluble carrier to produce the peptide. This solid-phase approach is generally described by Merrifield, et al. (1963) in ~
Am. Chem. Soc. 85:2149-2156, which is incorporated herein by reference.
The prepared protein and fragments thereof can be isolated and purified from the reaction mixture by means of peptide separation, e.g., by extraction, precipitation, electrophoresis, various forms of chromatography, and the like. The receptors of this invention can be obtained in varying degrees of purity depending upon desired uses. Purification can be accomplished by use of the protein purification techniques disclosed herein, see below, or by the use of the antibodies herein described in methods of immunoabsorbant affinity chromatography. This immunoabsorbant affinity chromatography is carried out by first linking the antibodies to a solid support and then contacting the linked antibodies with solubilized lysates of appropriate cells, lysates of other cells expressing the receptor, or lysates or supernatants of cells producing the polypeptide as a result of DNA techniques, see below.
Generally, the purified protein will be at least ..
about 40~ pure, ordinarily at least about 50~ pure, usually at least about 60~ pure, typically at least about 70~ pure, more typically at least about 80~ pure, preferable at least about 90~ pure and more preferably at least about 95~ pure, and in particular embodiments, 97~-99~ or more. Purity will usually be on a weight basis, but can also be on a molar basis. Different assays will 5 be applied as appropriate. Similar concepts apply'to polynucleotides and antibodies.
VI. Antibodies Antibodies can be raised to the various mammalian 10 IL-1RD8, IL-1RD9, or IL-1RD10 described herein, e.g., primate IL-1RD9 polypeptides and fragments thereof, both in naturally occurring native forms and in their recombinant forms, the difference being that antibodies to the active receptor are more likely to recognize 15 epitopes which are only present in the native conformations. Denatured antigen detection can also be useful in, e.g., Western analysis. Anti-idiotypic antibodies are also contemplated, which would be useful as agonists or antagonists of a natural receptor or an 20 antibody.
Antibodies, including binding fragments and single chain versions, against predetermined fragments of the polypeptide can be raised by immunization of animals with conjugates of the fragments with immunogenic proteins.
25 Monoclonal antibodies are prepared from cells secreting the desired antibody. These antibodies can be screened for binding to normal or defective protein, or screened for agonistic or antagonistic activity. These monoclonal antibodies will usually bind with at least a KD of about 30 1 mM, more usually at least about 300 EtM, typically at least about 100~.1M, more typically at least about 30 E,IM, preferably at least about 10 E.lM, and more preferably at least about 3 E1M or better.
The antibodies, including antigen binding fragments, 35 of this invention can have significant diagnostic or therapeutic value. They can be potent antagonists that bind to the receptor and inhibit binding to ligand or ., inhibit the ability of the receptor to elicit a biological response, e.g., act on its substrate. They also can be useful as non-neutralizing antibodies and can be coupled to toxins or radionuclides to bind producing cells, or cells localized to the source of the interleukin. Further, these antibodies can be conjugated to drugs or other therapeutic agents, either direct3y or indirectly by means of a linker.
The antibodies of this invention can also be useful in diagnostic applications. As capture or non-neutralizing antibodies, they might bind to the receptor without inhibiting ligand or substrate binding.
As neutralizing antibodies, they can be useful in competitive binding assays. They will also be useful in detecting or quantifying ligand. They may be used as reagents for Western blot analysis, or for immunoprecipitation or immunopurification of the respective protein.
Protein fragments may be joined to other materials, particularly polypeptides, as fused or covalently joined polypeptides to be used as immunogens. Mammalian IL-1Rs and fragments may be fused or covalently linked to a variety of immunogens, such as keyhole limpet hemocyanin, bovine serum albumin, tetanus toxoid, etc. See Microbiology, Hoeber Medical Division, Harper and Row, 1969; Landsteiner (1962) specificity of Serological ~teactions, Dover Publications, New York; and Williams, et al . ( 1967 ) Methods in Immunolocrv and Immunochemistrv, Vol. 1, Academic Press, New York; each of which are incorporated herein by reference, for descriptions of methods of preparing polyclonal antisera. A typical method involves hyperimmunization of an animal with an antigen. The blood of the animal is then collected shortly after the repeated immunizations and the gamma globulin is isolated.
In some instances, it is desirable to prepare monoclonal antibodies from various mammalian hosts, such as mice, rodents, primates, humans, etc. Description of techniques for preparing such monoclonal antibodies may be found in, e.g., Stites, et al. (eds.) $asic and Clinical Immunology (4th ed.), Lange Medical Publications, Los Altos, CA, and references cited therein; Harlow and Lane (1988) Antibodies- A Laboratory nual, CSH Press; Goding (1986) Monoclonal Antibodies-Principles and Practice (2d ed.) Academic Press, New York; and particularly in Kohler and Milstein (19'7S~ in ture 256:495-497, which discusses one method of generating monoclonal antibodies. Each of these references is incorporated herein by reference.
Summarized briefly, this method involves injecting an animal with an immunogen. The animal is then sacrificed and cells taken from its spleen, which are then fused with myeloma cells. The result is a hybrid cell or "hybridoma" that is capable of reproducing in vitro. The population of hybridomas is then screened to isolate individual clones, each of which secrete a single antibody species to the immunogen. In this manner, the individual antibody species obtained are the products of immortalized and cloned single B cells from the immune animal generated in response to a specific site recognized on the immunogenic substance.
Other suitable techniques involve in vitro exposure of lymphocytes to the antigenic polypeptides or alternatively to selection of libraries of antibodies in phage or similar vectors. See, Huse, et al. (1989) "Generation of a Large Combinatorial Library of the Immunoglobulin Repertoire in Phage Lambda," Science 246:1275-1281; and Ward, et al. (1989) Nature 341:544-546, each of which is hereby incorporated herein by reference. The polypeptides and antibodies of the present invention may be used with or without modification, including chimeric or humanized antibodies.
Frequently, the polypeptides and antibodies will be labeled by joining, either covalently or non-covalently, a substance which provides for a detectable signal. A
wide variety of labels and conjugation techniques are known and are reported extensively in both the scientific and patent literature. Suitable labels include radionuclides, enzymes, substrates, cofactors, inhibitors, fluorescent moieties, chemiluminescent 48 _ moieties, magnetic particles, and the like. Patents, teaching the use of such labels include U.S. Patent Nos.
3,817,837; 3,850,752; 3,939,350; 3,996,345; 4,277,437.;
4,275,149; and 4,366,241. Also, recombinant or chimeric immunoglobulins may be produced, see Cabilly, U.S~ Patent No. 4,816,567; or made in transgenic mice, see Mendez, et al. (1997) Nature Genetics 15:146-256. These references are incorporated herein by reference.
The antibodies of this invention can also be used for affinity chromatography in isolating the IL-lRs., Columns can be prepared where the antibodies are linked to a solid support, e.g., particles, such as agarose, Sephadex, or the like, where a cell lysate may be passed through the column, the column washed, followed by increasing concentrations of a mild denaturant, whereby the purified protein will be released. The protein may be used to purify antibody.
The antibodies may also be used to screen expression libraries for particular expression products. Usually the antibodies used in such a procedure will be labeled with a moiety allowing easy detection of presence of antigen by antibody binding.
Antibodies raised against an IL-1R will also be used to raise anti-idiotypic antibodies. These will be useful in detecting or diagnosing various immunological conditions related to expression of the protein or cells which express the protein. They also will be useful as agonists or antagonists of the ligand, which may be competitive inhibitors or substitutes for naturally occurring ligands.
An IL-1R polypeptide that specifically binds to or that is specifically immunoreactive with an antibody generated against a defined immunogen, such as an immunogen consisting of the amino acid sequence of, e.g., SEQ ID NO: 4, 10, or 20, is typically determined in an immunoassay. The immunoassay typically uses a polyclonal antiserum which was raised, e.g., to a polypeptide of SEQ
ID NO: 4, 10, or 20. This antiserum is selected to have low crossreactivity against other IL-1R family members, e.g., IL-1Rs D1 through D8, preferably from the same species, and any such crossreactivity is removed by immunoabsorption prior to use in the immunoassay.
To produce antisera for use in an immunoassay, the polypeptide of, e.g., SEQ ID NO: 4, 10, or 20, is"
isolated as described herein. For example, recombinant polypeptide may be produced in a mammalian cell line. An appropriate host, e.g., an inbred strain of mice such as Balb/c, is immunized with the selected protein, typically using a standard adjuvant, such as Freund~s adjuvant, and a standard mouse immunization protocol (see Harlow and Lane, supra). Alternatively, a synthetic peptide derived from the sequences disclosed herein and conjugated to a carrier polypeptide can be used an immunogen. Polyclonal sera are collected and titered against the immunogen polypeptide in an immunoassay, e.g., a solid phase immunoassay with the immunogen immobilized on a solid support. Polyclonal antisera with a titer of 104 or greater are selected and tested for their cross reactivity against other IL-1R family members, e.g., IL-1RD1 through IL-1RD6, using a competitive binding immunoassay such as the one described in Harlow and Lane, supra, at pages 570-573. Preferably at least two IL-1R
family members are used in this determination. These IL-1R family members can be produced as recombinant polypeptides and isolated using standard molecular biology and protein chemistry techniques as described herein.
Immunoassays in the competitive binding format can be used for the crossreactivity determinations. For example, the polypeptide of SEQ ID NO: 4, 10, or 20 can be immobilized to a solid support. Polypeptides added to the assay compete with the binding of the antisera to the immobilized antigen. The ability of the above polypeptides to compete with the binding of the antisera to the immobilized polypeptide is compared to the polypeptides of IL-1RD1 through IL-1RD6. The percent crossreactivity for the above polypeptides is calculated, using standard calculations. Those antisera with less than 10~ crossreactivity with each of the polypeptides listed above are selected and pooled. The cross-reacting antibodies are then removed from the pooled antisera by immunoabsorption with the above-listed proteins.
5 The immunoabsorbed and pooled antisera are then used in a competitive binding immunoassay as described above to compare a second polypeptide to the immunogen polypeptide (e. g., the IL-1RD8, IL-1RD9, or IL-1RD10 like polypeptide of SEQ ID NO: 4, 10, or 20). To make this 10 comparison, the two polypeptides are each assayed at~a wide range of concentrations and the amount of each polypeptide required to inhibit 50~ of the binding of the antisera to the immobilized polypeptide is determined.
If the amount of the second polypeptide required is less 15 than twice the amount of the polypeptide of the selected polypeptide or polypeptides that is required, then the second polypeptide is said to specifically bind to an antibody generated to the immunogen.
It is understood that these IL-1R polypeptides are 20 members of a family of homologous polypeptides that comprise at least 7 genes previously identified. For a particular gene product, such as, e.g., IL-1RD9, the term refers not only to the amino acid sequences disclosed herein, but also to other polypeptides that are allelic, 25 non-allelic, or species variants. It is also understood that the terms include nonnatural mutations introduced by deliberate mutation using conventional recombinant technology such as single site mutation, or by excising short sections of DNA encoding the respective proteins, 30 or by substituting new amino acids, or adding new amino acids. Such minor alterations typically will substantially maintain the immunoidentity of the original molecule and/or its biological activity. Thus, these alterations include polypeptides that are specifically 35 immunoreactive with a designated naturally occurring IL-1RD8, IL-1RD9, or IL-1RD10 protein. The biological properties of the altered polypeptides can be determined by expressing the polypeptide in an appropriate cell line and measuring the appropriate effect, e.g., upon transfected lymphocytes. Particular polypeptide modifications considered minor would include conservative substitution of amino acids with similar chemical properties, as described above for the IL-1R family as a whole. By aligning a polypeptide optimally with the polypeptide of the IL-1Rs and by using the conventional immunoassays described herein to determine immunoidentity, one can determine the polypeptide compositions of the invention.
VII. Kits and quantitation Both naturally occurring and recombinant forms of the IL-1R like molecules of this invention are particularly useful in kits and assay methods. For example, these methods would also be applied to screening for binding activity, e.g., ligands for these proteins.
Several methods of automating assays have been developed in recent years so as to permit screening of tens of thousands of compounds per year. See, e.g., a BIOMEK
automated workstation, Beckman Instruments, Palo Alto, California, and Fodor, et al. (1991) Science 251:767-773, which is incorporated herein by reference. The latter describes means for testing binding by a plurality of defined polymers synthesized on a solid substrate. The development of suitable assays to screen for a ligand or agonist/antagonist homologous polypeptides can be greatly facilitated by the availability of large amounts of purified, soluble IL-1Rs in an active state such as is provided by this invention.
~ Purified IL-1RD8, IL-1RD9, or IL-1RD10 can be coated directly onto plates for use in the aforementioned ligand screening techniques. However, non-neutralizing antibodies to these polypeptides can be used as capture antibodies to immobilize the respective receptor on the solid phase, useful, e.g., in diagnostic uses.
This invention also contemplates use of IL-1RD8, IL-1RD9, or IL-1RD10 fragments thereof, peptides, and their fusion products in a variety of diagnostic kits and methods for detecting the presence of the protein or its 52 _ ligand. Alternatively, or additionally, antibodies against the molecules may be incorporated into the kits and methods. Typically the kit will have a compartment containing, e.g., either an IL-1RD9 peptide or gene segment or a reagent which recognizes one or the other.
Typically, recognition reagents, in the case of peptide, would be a ligand or antibody, or in the case of a gene segment, would usually be a hybridization probe.
A preferred kit for determining the concentration of IL-1RD8, IL-1RD9, or IL-1RD10 in a sample would typically comprise a labeled compound, e.g., ligand or antibody, having known binding affinity for IL-1RD9, a source of IL-1RD9 (naturally occurring or recombinant) as a positive control, and a means for separating the bound from free labeled compound, for example a solid phase for immobilizing the IL-1RD9 in the test sample.
Compartments containing reagents, and instructions, will normally be provided.
Antibodies, including antigen binding fragments, specific for mammalian IL-1RD8 or a peptide fragment, or receptor fragments are useful in diagnostic applications to detect the presence of elevated levels of ligand and/or its fragments. Diagnostic assays may be homogeneous (without a separation step between free reagent and antibody-antigen complex) or heterogeneous (with a separation step). Various commercial assays exist, such as radioimmunoassay (RIA), enzyme-linked immunosorbent assay (ELISA), enzyme immunoassay (EIA), enzyme-multiplied immunoassay technique (EMIT), substrate-labeled fluorescent immunoassay (SLFIA) and the like. For example, unlabeled antibodies can be employed by using a second antibody which is labeled and which recognizes the antibody to an IL-1R or to a particular fragment thereof. These assays have also been extensively discussed in the literature. See, e.g., Harlow and Lane (1988) Antibodies: A Laboratory Manual, CSH., and Coligan (ed. 1991) and periodic supplements, ('»rrent Protocols In Immunoloav Greene/Wiley, New York.

Anti-idiotypic antibodies may have similar use to serve as agonists or antagonists of IL-lRs. These should be useful as therapeutic reagents under appropriate circumstances.
Frequently, the reagents for diagnostic assays are supplied in kits, so as to optimize the sensitivity of the assay. For the subject invention, depending upon the nature of the assay, the protocol, and the label, either labeled or unlabeled antibody, or labeled ligand is , provided. This is usually in conjunction with other additives, such as buffers, stabilizers, materials necessary for signal production such as substrates for enzymes, and the like. Preferably, the kit will also contain instructions for proper use and disposal of the contents after use. Typically the kit has compartments for each useful reagent, and will contain instructions for proper use and disposal of reagents. Desirably, the reagents are provided as a dry lyophilized powder, where the reagents may be reconstituted in an aqueous medium having appropriate concentrations for performing the assay.
The aforementioned constituents of the diagnostic assays may be used without modification or may be modified in a variety of ways. For example, labeling may be achieved by covalently or non-covalently joining a moiety which directly or indirectly provides a detectable signal. In many of these assays, a test compound, IL-1R, or antibodies thereto can be labeled either directly or indirectly. Possibilities for direct labeling include label groups: radiolabels such as 1251, enzymes (U. S.
Pat. No. 3,645,090) such as peroxidase and alkaline phosphatase, and fluorescent labels (U.S. Pat. No.
3,940,475) capable of monitoring the change in fluorescence intensity, wavelength shift, or fluorescence polarization. Both of the patents are incorporated herein by reference. Possibilities for indirect labeling include biotinylation of one constituent followed by binding to avidin coupled to one of the above label ' groups.

54 _ There are also numerous methods of separating the bound from the free ligand, or alternatively the bound from the free test compound. The IL-1R can be immobilized on various matrixes followed by washing.
Suitable matrices include plastic such as an ELISA plate, filters, and beads. Methods of immobilizing the receptor to a matrix include, without limitation, direct adhesion to plastic, use of a capture antibody, chemical coupling, and biotin-avidin. The last step in this approach involves the precipitation of antibody/antigen complex by any of several methods including those utilizing, e.g., an organic solvent such as polyethylene glycol or a salt such as ammonium sulfate. Other suitable separation techniques include, without limitation, the fluorescein antibody magnetizable particle method described in Rattle, et al. (1984) Clin. Chem. 30(9):1457-1461, and the double antibody magnetic particle separation as described in U.S. Pat. No. 4,659,678, each of which is incorporated herein by reference.
The methods for linking protein or fragments to various labels have been extensively reported in the literature and do not require detailed discussion here.
Many of the techniques involve the use of activated carboxyl groups either through the use of carbodiimide or active esters to form peptide bonds, the formation of thioethers by reaction of a mercapto group with an activated halogen such as chloroacetyl, or an activated olefin such as maleimide, for linkage, or the like.
Fusion polypeptides will also find use in these applications.
Another diagnostic aspect of this invention involves use of oligonucleotide or polynucleotide sequences taken from the sequence of an IL-1R. These sequences can be used as probes for detecting levels of the respective IL-1R in patients suspected of having an immunological disorder. The preparation of both RNA and DNA nucleotide sequences, the labeling of the sequences, and the preferred size of the sequences has received ample description and discussion in the literature. Normally an oligonucleotide probe should have at least about 14 nucleotides, usually at least about 18 nucleotides, and the polynucleotide probes may be up to several kilobases.
Various labels may be employed, most commonly radionuclides, particularly 32P. However, other w techniques may also be employed, such as using biotin modified nucleotides for introduction into a polynucleotide. The biotin then serves as the site for binding to avidin or antibodies, which may be labeled with a wide variety of labels, such as radionuclides, fluorescers, enzymes, or the like. Alternatively, antibodies may be employed which can recognize specific duplexes, including DNA duplexes, RNA duplexes, DNA-RNA
hybrid duplexes, or DNA-protein duplexes. The antibodies in turn may be labeled and the assay carried out where the duplex is bound to a surface, so that upon the formation of duplex on the surface, the presence of antibody bound to the duplex can be detected. The use of probes to the novel anti-sense RNA may be carried out in conventional techniques such as nucleic acid hybridization, plus and minus screening, recombinational probing, hybrid released translation (HRT), and hybrid arrested translation (HART). This also includes amplification techniques such as polymerase chain reaction (PCR).
Diagnostic kits which also test for the qualitative or quantitative presence of other markers are also contemplated. Diagnosis or prognosis may depend on the combination of multiple indications used as markers.
Thus, kits may test for combinations of markers. See, e.g., Viallet, et al. (1989) Pr~arPSS in Growth Factor Res. 1:89-97.
VIII. Therapeutic Utility This invention provides reagents with significant therapeutic value. The IL-1Rs (naturally occurring or..
recombinant), fragments thereof, mutein receptors, and antibodies, along with compounds identified as having binding affinity to the receptors or antibodies, should be useful in the treatment of conditions exhibiting abnormal expression of the receptors of their ligands.
Such abnormality will typically be manifested by immunological disorders. Additionally, this invention should provide therapeutic value in various diseases or disorders associated with abnormal expression or abnormal triggering of response to the ligand. The IL-1 ligands have been suggested to be involved in morphologic development, e.g., dorso-ventral polarity determination, and immune responses, particularly the primitive innate responses. See, e.g., Sun, et al. (1991) Eur. J.
Hiochem. 196:247-254; Hultmark (1994) Nature 367:116-117.
Recombinant IL-lRs, muteins, agonist or antagonist antibodies thereto, or antibodies can be purified and then administered to a patient. These reagents can be combined for therapeutic use with additional active ingredients, e.g., in conventional pharmaceutically acceptable carriers or diluents, along with physiologically innocuous stabilizers and excipients.
These combinations can be sterile, e.g., filtered, and placed into dosage forms as by lyophilization in dosage vials or storage in stabilized aqueous preparations.
This invention also contemplates use of antibodies or binding fragments thereof which are not complement binding.
Ligand screening using IL-1R or fragments thereof can be performed to identify molecules having binding affinity to the receptors. Subsequent biological assays can then be utilized to determine if a putative ligand can provide competitive binding, which can block intrinsic stimulating activity. Receptor fragments can be used as a blocker or antagonist in that it blocks the activity of ligand. Likewise, a compound having intrinsic stimulating activity can activate the receptor and is thus an agonist in that it simulates the activity of ligand, e.g., inducing signaling. This invention further contemplates the therapeutic use of antibodies to IL-1Rs as antagonists.

The quantities of reagents necessary for effective therapy will depend upon many different factors, including means of administration, target site, reagent physiological life, pharmacological life, physiological state of the patient, and other medicants administered.
Thus, treatment dosages should be titrated to optimize safety and efficacy. Typically, dosages used in vitro may provide useful guidance in the amounts useful for in situ administration of these reagents. Animal testing of effective doses for treatment of particular disorders will provide further predictive indication of human dosage. Various considerations are described, e.g., in Gilman, et al. (eds., 1990) Goodman and Gilman's: The Pharmacological Bases of Therapeutics, 8th Ed., Pergamon Press; and Reminqton's Pharmaceutical Sciences, 17th ed.
(1990), Mack Publishing Co., Easton, Penn.; each of which is hereby incorporated herein by reference. Methods for administration are discussed therein and below, e.g., for oral, intravenous, intraperitoneal, or intramuscular administration, transdermal diffusion, and others.
Pharmaceutically acceptable carriers will include water, saline, buffers, and other compounds described, e.g., in the Merck Index, Merck & Co., Rahway, New Jersey.
Because of the likely high affinity binding, or turnover numbers, between a putative ligand and its receptors, low dosages of these reagents would be initially expected to be effective. And the signaling pathway suggests extremely low amounts of ligand may have effect. Thus, dosage ranges would ordinarily be expected to be in amounts lower than 1 mM concentrations, typically less than about 10 ).1M concentrations, usually less than about 100 nM, preferably less than about 10 pM (picomolar), and most preferably less than about 1 fM (ferntomolar), with an appropriate carrier. Slow release formulations, or slow release apparatus will often be utilized for continuous administration.
IL-lRs,~fragments thereof, and antibodies or its fragments, antagonists, and agonists, may be administered directly to the host to be treated or, depending on the size of the compounds, it may be desirable to conjugate them to carrier proteins such as ovalbumin or serum albumin prior to their administration. Therapeutic formulations may be administered in many conventional dosage formulations. 44Thile it is possible for the active ingredient to be administered alone, it is preferable to present it as a pharmaceutical formulation. Formulations comprise at least one active ingredient, as defined above, together with one or more acceptable carriers thereof. Each carrier must be both pharmaceutically~and physiologically acceptable in the sense of being compatible with the other ingredients and not injurious to the patient. Formulations include those suitable for oral, rectal, nasal, or parenteral (including subcutaneous, intramuscular, intravenous and intradermal) administration. The formulations may conveniently be presented in unit dosage form and may be prepared by methods well known in the art of pharmacy. See, e.g., Gilman, et al. (eds. 1990) Goodman and Gilman's: The Pl~armacolog~ical Bases of Therapeutics, 8th Ed., Pergamon Press; and Remincrton's Pharmaceutical Sciences, 17th ed.
(1990), Mack Publishing Co., Easton, Penn.; Avis, et al.
(eds. 1993) Pharmaceutical Dosacxe Forms: Parenteral Medications Dekker, NY; Lieberman, et al. (eds. 1990) Pharmaceutical Dosaae Forms: Tablets Dekker, NY; and Lieberman, et al. (eds. 1990) Pharmaceutical Dosage Forms: Disperse Svste~~ Dekker, NY. The therapy of this invention may be combined with or used in association with other therapeutic agents, particularly agonists or antagonists of other IL-1 family members.
IX. Ligands The description of the IL-1 receptors herein provide means to identify ligands, as described above. Such ligand should bind specifically to the respective receptor with reasonably high affinity. Typical ligand receptor binding constants will be at least about 30 mM, e.g., generally at least about 3 mM, more generally at least about 300 EtM, typically at least about 30 E.tM, 3 ~.LM, 300 nM, 30 nM, etc. Various constructs are made available which allow either labeling of the receptor to detect its ligand. For example, directly labeling IL-1R, fusing onto it markers for secondary labeling, e.g., FLAG
or other epitope tags, etc., will allow detection of receptor. This can be histological, as an affinity method for biochemical purification, or labeling or selection in an expression cloning approach. A two-hybrid selection system may also be applied making appropriate constructs with the available IL-1R
sequences. See, e.g., Fields and Song (1989) Natu~g~
340:245-246.
Generally, descriptions of IL-1Rs will be analogously applicable to individual specific embodiments directed to IL-1RD8, IL-1RD9, OR IL-1RD10 reagents and compositions.
The broad scope of this invention is best understood with reference to the following examples, which are not intended to limit the inventions to the specific embodiments.
EXAMPLES
I. General Methods Some of the standard methods are described or referenced, e.g., in Maniatis, et al. (1982) Molecular Cloning, A Laboratory Manual, Cold Spring Harbor Laboratory, Cold Spring Harbor Press; Sambrook, et al.
(1989) Molecular Clonincr: A Laboratory Manual, (2d ed.), vols. 1-3, CSH Press, NY; Ausubel, et al. Giolocrv Greene Publishing Associates, Brooklyn, NY; or Ausubel, et al.
(1987 and Supplements) Current Protocols in Molecular F~ioloav, Greene/Wiley, New York. Methods for protein purification include such methods as ammonium sulfate precipitation, column chromatography, electrophoresis, centrifugation, crystallization, and others. See, e.g., Ausubel, et al. (1987 and periodic supplements); Coligan, et al. (ed. 1996 and periodic supplements) Current Protocols In Protein Science Greene/Wiley, New York;
Deutscher (1990) "Guide to Protein Purification" in Met ods in Enzvmoloav, vol. 182, and other volumes in this series; and manufacturer's literature on use of protein purification products, e.g., Pharmacia, Piscataway, N.J., or Bio-Rad, Richmond, CA. Combination with recombinant techniques allow fusion to appropriate 5 segments, e.g., to a FLAG sequence or an eguivalent"which can be fused via a protease-removable sequence. See, e.g., Hochuli (1989) Chemische Industrie 12:69-70;
Hochuli (1990) "Purification of Recombinant Proteins with Metal Chelate Absorbent" in Setlow (ed.) Genetic 10 Enaineering Principle and Methods 12:87-98, Plenum Press, N.Y.; and Crowe, et al. (1992) QTAe~res~: The bah Level Exuression & Protein Purification Svstem QUIAGEN, Inc., Chatsworth, CA.
Computer sequence analysis is performed, e.g., using 15 available software programs, including those from the GCG
(U. Wisconsin) and GenBank sources. Public sequence databases were also used, e.g., from GenBank, NCBI, SWISSPROT, and others.
Many techniques applicable to IL-10 receptors may be 20 applied to IL-lRs, as described, e.g., in USSN 08/110,683 (IL-10 receptor), which is incorporated herein by reference for all purposes. Also, while many of the techniques described are directed to the IL-1RD9 reagents, corresponding methods will typically be 25 applicable with the IL-1RD8, and IL-1RD10 reagents. See also, USSN 60/065,776, filed November 17, 1997, and USSN
60/078,008, filed March 12, 1998, both of which are incorporated herein by reference.
30 II. Computational Analysis.
Human sequences related to IL-1Rs were identified from various EST databases using, e.g., the BLAST server (Altschul, et al. (1994) Nature Genet. 6:119-129). More sensitive pattern- and profile-based methods (Bork and 35 Gibson (1996) Meth. Enzvmol. 266:162-184) were used to identify a fragment of a gene which exhibited certain homology to the IL-lRs.
III. Cloning of full-length human IL-1R cDNAs.

PCR primers derived from the IL-1RD8, IL-1RD9, or IL-1RD10 sequences are used (Nomura, et al. (1994) p ,es. 1:27-35) to probe an appropriate human cDNA library to yield a full length IL-1RD9 or IL-1RD10 cDNA sequence or to probe a human erythroleukemic, TF-1 cell line=
derived cDNA library (Kitamura, et al. (1989) Elood 73:375-380) to yield the IL-1R8 cDNA sequence. Full length cDNAs for human IL-1RD9 are cloned, e.g., by DNA
hybridization screening of agtl0 phage. PCR reactions were conducted using T. aquaticus Taqplus DNA polymerase (Stratagene) under appropriate conditions.
IV. Localization of IL-1RD8, IL-1RD9, and IL-1RD10 mRNA
Human multiple tissue (Cat# l, 2) and cancer cell line blots (Cat# 7757-1), containing approximately 2 ~..l,g of poly(A)+ RNA per lane, are purchased from Clontech (Palo Alto, CA). Probes are radiolabeled with [a-32p~
dATP, e.g., using the Amersham Rediprime random primer labeling kit (RPN1633). Prehybridization and hybridizations are performed at 65° C in 0.5 M Na2HP04, 7~ SDS, 0.5 M EDTA (pH 8.0). High stringency washes are conducted, e.g., at 65° C with two initial washes in 2 x SSC, 0.1~ SDS for 40 min followed by a subsequent wash in 0.1 x SSC, 0.1~ SDS for 20 min. Membranes are then exposed at -70° C to X-Ray film (Kodak) in the presence of intensifying screens. More detailed studies by cDNA
library Southerns are performed with selected human IL-1RD9 clones to examine their expression in hemopoietic or other cell subsets.
Two prediction algorithms that take advantage of the patterns of conservation and variation in multiply aligned sequences, PHD (Rost and Sander (1994) Proteins 19:55-72) and DSC (King and Sternberg (1996) Protein Sci.
5:2298-2310), are used.
Alternatively, two appropriate primers are selected from Tables l, 2, or 3. RT-PCR is used on an appropriate mRNA sample selected for the presence of message to produce a cDNA, e.g., a sample which expresses the gene.

Full length clones may be isolated by hybridization of cDNA libraries from appropriate tissues pre-selected by PCR signal. Northern blots can be performed.
Message for genes encoding, e.g., IL-1RD9 will be assayed by appropriate technology, e.g., PCR, - "
immunoassay, hybridization, or otherwise. Tissue and organ cDNA preparations are available, e.g., from Clontech, Mountain View, CA. Identification of sources of natural expression are useful, as described. And the identification of functional receptor subunit pairings will allow for prediction of what cells express the combination of receptor subunits which will result in a physiological responsiveness to each of the IL-1 ligands.
The message for IL-IRD9 is quite rare, as it is not found with a degree of frequency in the available sequence databases. This suggests, e.g., a very rare message, or a highly restricted distribution. IL-1R9 is expressed predominantly on T cells, NK cells, monocytes and dendritic cells.
Southern Analysis on cDNA libraries can be performed:
DNA (5 ~,g) from a primary amplified cDNA library is digested with appropriate restriction enzymes to release the inserts, run on a 1~ agarose gel and transferred to a nylon membrane' (Schleicher and Schuell, Keene, NH).
Samples for human mRNA isolation may include, e.g..
peripheral blood mononuclear cells (monocytes, T cells, NK cells, granulocytes, B cells), resting (T100);
peripheral blood mononuclear cells, activated with anti-CD3 for 2, 6, 12 h pooled (T101); T cell, THO clone Mot 72, resting (T102); T cell, THO clone Mot 72, activated with anti-CD28 and anti-CD3 for 3, 6, 12 h pooled (T103);
T cell, TH0 clone Mot 72, anergic treated with specific peptide for 2, 7, 12 h pooled (T104); T cell, TH1 clone HY06, resting (T107); T cell, TH1 clone HY06, activated with anti-CD28 and anti-CD3 for 3, 6, 12 h pooled (T108);
T cell, TH1 clone HY06, anergic treated with specific peptide for 2, 6, 12 h pooled (T109); T cell, TH2 clone HY935, resting (T110); T cell, TH2 clone HY935, activated with anti-CD28 and anti-CD3 for 2, 7, 12 h pooled (T111);

T cells CD4+CD45R0- T cells polarized 27 days in anti-CD28, IL-4, and anti IFN-y, TH2 polarized, activated with anti-CD3 and anti-CD28 4 h (T116); T cell tumor lines Jurkat and Hut78, resting (T117); T cell clones, pooled AD130.2, Tc783.12, Tc783.13, Tc783.58, Tc782.69, resting (T118); T cell random 'y8 T cell clones, resting {T119);
Splenocytes, resting {B100); Splenocytes, activated with anti-CD40 and IL-4 (B101); B cell EBV lines pooled WT49, RSB, JY, CVIR, 721.221, RM3, HSY, resting {B102); B cell line JY, activated with PMA and ionomycin for 1, 6 h~
pooled (B103); NK 20 clones pooled, resting {K100); NK 20 clones pooled, activated with PMA and ionomycin for 6 h (K101); NKL clone, derived from peripheral blood of LGL
leukemia patient, IL-2 treated {K106); NK cytotoxic clone 640-A30-l, resting (K107); hematopoietic precursor line TF1, activated with PMA and ionomycin for 1, 6 h pooled (C100); U937 premonocytic line, resting (M100); U937 premonocytic line, activated with PMA and ionomycin for 1, 6 h pooled (M101); elutriated monocytes, activated with LPS, IFN~y, anti-IL-10 for 1, 2, 6, 12, 24 h pooled (M102); elutriated monocytes, activated with LPS, IFN~y, IL-10 for 1, 2, 6, 12, 24 h pooled (M103); elutriated monocytes, activated with LPS, IFN~y, anti-IL-10 for 4, 16 h pooled {M106); elutriated monocytes, activated with LPS, IFNy, IL-10 for 4, 16 h pooled (M107); elutriated monocytes, activated LPS for 1 h (M108); elutriated monocytes, activated LPS for 6 h {M109); DC 70~ CDla+, from CD34+ GM-CSF, TNFa 12 days, resting (D101); DC 70~
CDla+, from CD34+ GM-CSF, TNFa 12 days, activated with PMA and ionomycin for 1 hr {D102); DC 70~ CDIa+, from CD34+ GM-CSF, TNFa 12 days, activated with PMA and ionomycin for 6 hr (D103); DC 95~ CDla+, from CD34+ GM-CSF, TNFa 12 days FACS sorted, activated with PMA and ionomycin for 1, 6 h pooled (D104); DC 95~ CD14+, ex CD34+ GM-CSF, TNFa 12 days FRCS sorted, activated with PMA and ionomycin 1, 6 hr pooled {D105); DC CDla+ CD86+, from CD34+ GM-CSF, TNFa 12 days FAGS sorted, activated~~
with PMA and ionomycin for 1, 6 h pooled (D106); DC from monocytes GM-CSF, IL-4 5 days, resting (D107); DC from monocytes GM-CSF, IL-4 5 days, resting (D108); DC from monocytes GM-CSF, IL-4 5 days, activated LPS 4, 16 h pooled (D109); DC from monocytes GM-CSF, IL-4 5 days, activated TNFoc, monocyte supe for 4, 16 h pooled (D110);
leiomyoma L11 benign tumor (X101); normal myometrium M5 (0115); malignant leiomyosarcoma GS1 (X103); lung fibroblast sarcoma line MRC5, activated with PMA and ionomycin for 1, 6 h pooled (C101); kidney epithelial carcinoma cell line CHA, activated with PMA and ionomycin for 1, 6 h pooled (C102); kidney fetal 28 wk male (0100);
lung fetal 28 wk male (0101); liver fetal 28 wk male (0102); heart fetal 28 wk male (0103); brain fetal 28 wk male (0104); gallbladder fetal 28~wk male (0106); small intestine fetal 28 wk male (0107); adipose tissue fetal 28 wk male (0108); ovary fetal 25 wk female (0109);
uterus fetal 25 wk female (0110); testes fetal 28 wk male (0111); spleen fetal 28 wk male (0112); adult placenta 28 wk (0113); and tonsil inflamed, from 12 year old (X100);
psoriasis human skin sample; normal human skin sample;
pool of rheumatioid arthritis human; Hashimoto's thryroiditis thryroid; normal human throid; ulceratived colitis human colon; normal human colon; normal weight monkey colon; pheumocysitc carnii pneumonia lung;
allergic lung; pool of three heavy smoker human lung;
pool of two normal human lung; Ascaris-challenged monkey lung, 24hr; Ascaris-challenged monkey lung, 4hr; normal weight monkey lung..
IL-1RD8 message is described below in Table 5.
There appears to be a correlation between developmental stage of tissues and the levels of messages: fetal and transformed tissues express high levels, whereas normal, adult tissues express low levels (with the exception of skeletal muscle). Further insights into this phenomenon will need further experiments.
Message for genes encoding IL-1RD8 will be assayed by appropriate technology, e.g., PCR, immunoassay, hybridization, or otherwise. Tissue and organ cDNA
preparations are available, e.g., from Clontech, Mountain View, CA. Identification of sources of natural expression are useful, as described. And the identification of functional receptor subunit pairings will allow for prediction of what cells express the combination of receptor subunits which will result in a 5 physiological responsiveness to each of the IL-1 ligands.
Table 5 Multiple Tissue Northern Blots screened with radiolabeled were a 10 probe, encompassing the cytoplasmic region of Interleukin-1 receptor R8 (IL-1RD8). The resu lts are summarized below:

In all cases listed there is a smaller band 3.4 Kb and at in a few cases a larger band at 4.0 Kb as well.

15 Tissue 3.4 kb 4.0 kb Spleen weak Thymus weak Prostate weak 2 Testis weak Ovary weak Small Intestine weak Colon (mucosal lining) weak Peripheral Blood Leukocyte weak 25 Heart moderate Brain weak Placenta moderate Lung weak Liver weak 3 Skeletal Muscle strong Kidney weak Pancreas weak Fetal brain strong weak Fetal lung strong weak 3 Fetal Liver strong weak Fetal Kidney strong weak proleukocytic leukemia HL-60 strong HeLa Cell S3 very strong weak 40 Chronic myelogenous leukemia, very strong weak Lymphoblastic leukemia, MOLT-4 weak Burkitt's lymphoma Rajii moderate Colorectal adenocarcinoma SW40 very strong strong Lung carcinoma A549 strong strong 45 Melanoma very strong weak V. Cloning of species counterparts of IL-lRDs 50 Various strategies are used to obtain species counterparts of IL-1RD8, IL-1RD9, and IL-1RD10 preferably from other primates. One method is by cross hybridization using closely related species DNA probes:
It may be useful to go into evolutionarily similar 55 species as intermediate steps. Another method is by using specific PCR primers based on the identification of blocks of similarity or difference between genes, e.g., areas of highly conserved or nonconserved polypeptide or nucleotide sequence. In addition, gene sequence databases may be screened for related sequences from other species.
VI. Production of mammalian IL-1RD8, IL-1RD9, and IL-1RD10 protein An appropriate, e.g., GST, fusion construct is engineered for expression, e.g., in E. coli. For example, a mouse IGIF pGex plasmid is constructed and transformed into E. coli. Freshly transformed cells are grown, e.g_., in LB medium containing 50 ~,g/ml ampicillin and induced with IPTG (Sigma, St. Louis, MO). After overnight induction, the bacteria are harvested and the pellets containing, e.g., the IL-1R8 polypeptide are isolated. The pellets are homogenized, e.g., in TE
buffer (50 mM Tris-base pH 8.0, 10 mM EDTA and 2 mM
pefabloc) in 2 liters. This material is passed through a microfluidizer (Microfluidics, Newton, MA) three times.
The fluidized supernatant is spun down on a Sorvall GS-3 rotor for 1 h at 13,000 rpm. The resulting supernatant containing the IL-1R polypeptide is filtered and passed over a glutathione-SEPHAROSE column equilibrated in 50 mM
Tris-base pH 8Ø The fractions containing the IL-1RD9-GST fusion protein are pooled and cleaved, e.g., with thrombin (Enzyme Research Laboratories, Inc., South Bend, IN). The cleaved pool is then passed over a Q-SEPHAROSE
column equilibrated in 50 mM Tris-base. Fractions containing IL-1RD9 are pooled and diluted in cold distilled H20, to lower the conductivity, and passed back over a fresh Q-Sepharose column, alone or in succession with an immunoaffinity antibody column.. Fractions containing the IL-1RD9 polypeptide are pooled, aliquoted, and stored in the -70° C freezer.
Comparison of the CD spectrum with IL-1R polypeptide may suggest that the protein is correctly folded. See.
Hazuda, et al. (1969) J. Biol. Chem. 264:1689-1693.

VII. Determining physiological forms of receptors The IL-1a and IL-1(3 ligands bind an IL-1RD1 as the primary receptor and this complex then forms a high affinity receptor complex with the IL-1RD3. Such receptor subunits are probably shared with the receptors for the new IL-1 ligand family members. See, e.g., USSN
60/044,165 and USSN 60/055,111. Combination of the IL-1RD9 (a subunit type, based upon sequence analysis) will combine with the IL-1RD5 ((3 subunit type, based upon sequence analysis) to form a heterodimer receptor. The IL-1b and IL-1E ligands each probably signal through a receptor comprising the association of IL-1RD4, IL-1RD6, or IL-1RD9 (alpha components) with IL-1RD3, IL-1RD8, or IL-1RD10 (beta components).
These defined subunit combinations can be tested now with the provided reagents. In particular, appropriate constructs can be made for transformation or transfection of subunits into cells. Constructs for the alpha chains, e.g., IL-1RD1, IL-1RD4, IL-1RD6, and IL-1RD9 forms can be made. Likewise for the beta subunits IL-1RD3, IL-1RD5, IL-1RD7, and IL-1RD8. Structurally, the IL-1RD10 is most similar to the IL-1RD8, suggesting that it may also be a beta receptor subunit. Combinatorial transfections of transformations can make cells expressing defined subunits, which can be tested for response to each of the IL-1 ligands. Appropriate cell types can be used, e.g., 293 T cells, Jurkat cells, with, e.g., a nuclear kappa B
lNFtcb)-controlled Iuciferase reporter construct such as described e.g., in Otieno et al.,(1997) Am J Phvsiol 273-xxx.
Such combinations of various IL-1 ligands and receptors were tested to determine if a functional signaling complex had been formed using an NFxb-controlled luciferase reporter construct to indicate formation of a functional signaling complex (+) or failure to form a functional signaling complex (-). The results, presented below, IL-1a + IL-1~ + IL-1RD1 + IL-1RD3 = +;

IL-1a + IL-1~i+IL-1RD1 + IL-1RD5 = +;
IL-1a + IL-1~i + IL-1RD1 + IL-1RD8 = +;
IL-1a + IL-1(3+ IL-1RD1 + IL-1RD10 may = +/3;
w suggest that IL-1RD3, IL-1RD5, IL-1RD8, and IL-1RD10 may functionally substitute for each other when in combination with IL-la + IL-1(3+ IL-1RD1.
Other combinations (below) demonstrate a failure of functional substitution; suggesting the importance o~
contextual dependence on substitution e.g., IL-1RD3, and IL-1RD8 cannot functionally replace IL-1RD5 in the following combination: IL-1'y+ IL-1RD9 + IL-1RD5.
IL-1'y+ IL-1RD9 + IL-1RD5 = +;
IL-1Y+IL-1RD9 + IL-1RD3 = -;
IL-1y + IL-1RD9 + IL-1RD8 = -;
A further series of experiments tested the ability of mouse (m) and human (h) homologues to functionally substitute for each other. The results, shown below, mIL-1~ + mIL-1RD5 + mIL-1RD9 = +;
mIL-1'y+ mIL-1RD5 + hIL-1RD9 = -;
mIL-1y+ hIL-1RD5 + hIL-1RD9 = -;
mIL-hy+ hIL-1RD5 + mIL-2RD9 = -;
hIL-1'y+ mIL-1RD5 + mIL-1RD9 = -;
hIL-1~+ mIL-1RD5 + hIL-1RD9 = -;
hIL-hy+ hIL-1RD5 + mIL-1RD9 = -;
hIL-1Y+ hIL-1RD5 + hIL-1RD9 = +;
suggest that species homogeneity is required to form a functioning complex in this particular constellation of ligand and receptor units.
Biological assays will generally be directed to the ligand binding feature of the protein or to the kinase/phosphatase activity of the receptor. The activity will typically be reversible, as are many other enzyme actions mediate phosphatase or phosphorylase 69 _ activities, which activities are easily measured by standard procedures. See, e.g., Hardie, et al. (eds.
1995) The Protein Kinase FactBook vols. I and II, Academic Press, San Diego, CA; Hanks, et al. (1991) Meth.
Enzvmol. 200:38-62; Hunter, et al. (1992) Cell 70~3~5-388; Lewin (1990) Cell 61:743-752; Pines, et al. (1991) Cold SRring~ Harbor Svmp. Ouant. Biol. 56:449-463; and Parker, et al. (1993) at re 363:736-738.
The family of interleukins 1 contains molecules, each of which is an important mediator of inflammatory disease. For a comprehensive review, see Dinarello (1996) "Biologic basis for interleukin-1 in disease"
Blood 87:2095-2147. There are suggestions that the various IL-1 ligands may play important roles in the initiation of disease, particularly inflammatory responses. The finding of novel polypeptides related to the IL-1 family furthers the identification of molecules that provide the molecular basis for initiation of disease and allow for the development of therapeutic strategies of increased range and efficacy.
VIII. Preparation of antibodies specific for IL-1Rs Inbred Balb/c mice are immunized intraperitoneally with recombinant forms of the polypeptide, e.g., purified IL-1RD8, IL-1RD9, and IL-1RD10, or stable transfected NIH-3T3 cells. Animals are boosted at appropriate time points with protein, with or without additional adjuvant, to further stimulate antibody production. Serum is collected, or hybridomas produced with harvested spleens.
Alternatively, Balb/c mice are immunized with cells transformed with the gene or fragments thereof, either endogenous or exogenous cells, or with isolated membranes enriched for expression of the antigen. Serum is collected at the appropriate time, typically after numerous further administrations. Various gene therapy techniques may be useful, e.g., in producing protein in situ, for generating an immune response.
Monoclonal antibodies may be made. For example, splenocytes are fused with an appropriate fusion partner and hybridomas are selected in growth medium by standard procedures. Hybridoma supernatants are screened for the presence of antibodies which bind to the desired IL-1R, e.g., by ELISA or other assay. Antibodies which 5 selectively recognize specific IL-1R embodiments may also be selected or prepared.
In another method, synthetic peptides or purified protein are presented to an immune system to generate monoclonal or polyclonal antibodies. See, e.g., Coligan 10 (1991) Current Protocols in Immunology Wiley/Greene;~and Harlow and Lane (1989) Antibodies: A Laboratory Manual Cold Spring Harbor Press. In appropriate situations, the binding reagent is either labeled as described above, e.g., fluorescence or otherwise, or immobilized to a 15 substrate for panning methods. Nucleic acids may also be introduced into cells in an animal to produce the antigen, which serves to elicit an immune response. See, e.g., Wang, et al. (1993) Proc. Nat'1. Acad. Sci.
90:4156-4160; Barry, et al. (1994) BioTechniaues 16:616-20 619; and Xiang, et al. (1995) Immunity 2:129-135.
Moreover, antibodies which may be useful to determine the combination of the IL-1RD8, IL-1RD9, or IL-1RD10 with a functional beta subunit may be generated.
Thus, e.g., epitopes characteristic of a particular 25 functional alpha/beta combination may be identified with appropriate antibodies.
IX. Production of fusion proteins with IL-1Rs Various fusion constructs are made with IL-lRs. A
30 portion of the appropriate gene is fused to an epitope tag, e.g., a FLAG tag, or to a two hybrid system construct. See, e.g., Fields and Song (1989) Nature 340:245-246.
The epitope tag may be used in an expression cloning 35 procedure with detection with anti-FLAG antibodies to detect a binding partner, e.g., ligand for the respective IL-1R. The two hybrid system may also be used to isolate proteins which specifically bind, e.g., to IL-1RD9.

X. Structure activity relationship Information on the criticality of particular residues is determined using standard procedures and analysis. Standard mutagenesis analysis is performed, e.g., by generating many different variants at determined positions, e.g., at the positions identified above, and evaluating biological activities of the variants. This may be performed to the extent of determining positions which modify activity, or to focus on specific positions to determine the residues which can be substituted to either retain, block, or modulate biological activity.
Alternatively, analysis of natural variants can indicate what positions tolerate natural mutations. This may result from population analysis of variation among individuals, or across strains or species. Samples from selected individuals are analyzed, e.g., by PCR analysis c and sequencing. This allows evaluation of population polymorphisms.
XI. Isolation of a ligand for IL-1Rs An IL-1R can be used as a specific binding reagent to identify its binding partner, by taking advantage of its specificity of binding, much like an antibody would be used. Typically, the binding receptor is a heterodimer of receptor subunits. A binding reagent is either labeled as described above, e.g., fluorescence or otherwise, or immobilized to a substrate for panning methods.
The binding composition is used to screen an expression library made from a cell line which expresses a binding partner, i.e., ligand, preferably membrane associated. Standard staining techniques are used to detect or sort surface expressed ligand, or surface expressing transformed cells are screened by panning.
Screening of intracellular expression is performed by various staining or immunofluorescence procedures. See also McMahan, et al. (1991) ~O J. 10:2821-2832.
For example, on day 0, precoat 2-chamber permanox slides with 1 ml per chamber of fibronectin, 10 ng/ml in PBS, for 30 min at room temperature. Rinse once with PBS. Then plate COS cells at 2-3 x 105 cells per chamber in 1.5 ml of growth media. Incubate overnight at 37' C.
On day 1 for each sample, prepare 0.5 ml of a solution of 66 ~.g/ml DEAF-dextran, 66 E.tM chloroquin'~, and 4 ~.l,g DNA in serum free DME. For each set, a positive control is prepared, e.g., of IL-1R-FLAG cDNA at 1 and 1/200 dilution, and a negative mock. Rinse cells with serum free DME. Add the DNA solution and incubate 5 hr at 37' C. Remove the medium and add 0.5 ml 10~ DMSO~in DME for 2.5 min. Remove and wash once with DME. Add 1.5 ml growth medium and incubate overnight.
On day 2, change the medium. On days 3 or 4, the cells are fixed and stained. Rinse the cells twice with Hank's Buffered Saline Solution (HBSS) and fix in 4~
paraformaldehyde (PFA)/glucose for 5 min. Wash 3X with HESS. The slides may be stored at -80' C after all liquid is re;noved. For each chamber, 0.5 ml incubations are performed as follows. Add HBSS/saponin (0.1~) with 32 ).~,1/ml of 1 M NaN3 for 20 min. Cells are then washed with HBSS/saponin 1X. Add appropriate IL-1R or IL-1R/antibody complex to cells and incubate for 30 min.
Wash cells twice with HBSS/saponin. If appropriate, add first antibody for 30 min. Add second antibody, e.g., Vector anti-mouse antibody, at 1/200 dilution, and incubate for 30 min. Prepare ELISA solution, e.g., Vector Elite ABC horseradish peroxidase solution, and preincubate for 30 min. Use, e.g., 1 drop of solution A
(avidin) and 1 drop solution B (biotin) per 2.5 ml HBSS/saponin. Wash cells twice with HBSS/saponin. Add ABC HRP solution and incubate for 30 min. Wash cells twice with HBSS, second wash for 2 min, which closes cells. Then add Vector diaminobenzoic acid (DAB) for 5 to 10 min. Use 2 drops of buffer plus 4 drops DAB plus 2 drops of H202 per 5 ml of glass distilled water.
Carefully remove chamber and rinse slide in water. Air dry for a few minutes, then add 1 drop of Crystal Mount and a cover slip. Bake for 5 min at 85-90' C.

Evaluate positive staining of,pools and progressively subclone to isolation of single genes responsible for the binding.
Alternatively, IL-1R reagents are used to affinity purify or sort out cells expressing a putative ligand.
See, e.g., Sambrook, et al. or Ausubel, et al.
Another strategy is to screen for a membrane bound receptor by panning. The receptor cDNA is constructed as described above. The ligand can be immobilized and used to immobilize expressing cells. Immobilization may be achieved by use of appropriate antibodies which recognize, e.g., a FLAG sequence of an IL-1R fusion construct, or by use of antibodies raised against the first antibodies. Recursive cycles of selection and amplification lead to enrichment of appropriate clones and eventual isolation of receptor expressing clones.
Phage expression libraries can be screened by mammalian IL-lRs. Appropriate label techniques, e.g., anti-FLAG antibodies, will allow specific labeling of appropriate clones.
Many modifications and variations of this invention can be made without departing from its spirit and scope, as will be apparent to those skilled in the art. The specific embodiments described herein are offered by way of example only, and the invention is to be limited by the terms of the appended claims, along with the full scope of equivalents to which such claims are entitled;
and the invention is not to be limited by the specific embodiments that have been presented herein by way of example.

SEQUENCE LISTING
(1) GENERAL INFORMATION:
(i) APPLICANT: Schering Corporation (ii) TITLE OF INVENTION: HUMAN RECEPTOR PROTEINS; RELATED
REAGENTS
(iii) NUMBER OF SEQUENCES: 33 (iv) CORRESPONDENCE ADDRESS:
(A) ADDRESSEE: Schering-Plough Corporation (B) STREET: 2000 Galloping Hill Road (C) CITY: Kenilworth (D) STATE: New Jersey (E) COUNTRY: USA
(F) ZIP: 07033-0530 (v) COMPUTER READABLE FORM:
(A) MEDIUM TYPE: Floppy disk (B) COMPUTER: IBM PC compatible (C) OPERATING SYSTEM: PC-DOS/MS-DOS
(D) SOFTWARE: PatentIn Release #1.0, Version #1.30 (vi) CURRENT APPLICATION DATA:
(A) APPLICATION NUMBER:
(B) FILING DATE:
(C) CLASSIFICATION:
(vii) PRIOR APPLICATION DATA:
(A) APPLICATION NUMBER: US 60/078,008 (B) FILING DATE: 12-MAR-1998 (vii) PRIOR APPLICATION DATA:
(A) APPLICATION NUMBER: US 60/081,883 (B) FILING DATE: 15-APR-1998 (vii) PRIOR APPLICATION DATA:
(A) APPLICATION NUMBER: US 60/095,987 (B) FILING DATE: 10-AUG-1998 (vii) PRIOR APPLICATION DATA:
(A) APPLICATION NUMBER: US 08/971,635 (B) FILING DATE: 17-NOV-1997 (vii) PRIOR APPLICATION DATA:
(A) APPLICATION NUMBER: US 09/040,714 (B) FILING DATE: 18-MAR-1998 (vii) PRIOR APPLICATION DATA:
(A) APPLICATION NUMBER: US 08/951,829 (B) FILING DATE: 15-OCT-1997 (ix) TELECOMMUNICATION INFORMATION:
(A) TELEPHONE: (908) 298-2135 (B) TELEFAX: (908) 298-5388 (2) INFORMATION FOR SEQ ID NO:1:

(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 1737 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii)~MOLECULE TYPE: cDNA
(iii) HYPOTHETICAL: YES
(ix) FEATURE:
(A) NAME/KEY: CDS
(B) LOCATION: 1..1737 (ix) FEATURE:
(A) NAME/KEY: misc-feature (B) LOCATION: 342..343 (D) OTHER INFORMATION: /note= "splice junction"
(ix) FEATURE:
(A) NAME/KEY: misc_feature (B) LOCATION: 453..454 (D) OTHER INFORMATION: /note= "splice junction"
(ix) FEATURE:
(A) NAME/KEY: misc_feature (B) LOCATION: 756..757 (D) OTHER INFORMATION: /note= "splice junction"
(ix) FEATURE:
(A) NAME/KEY: misc-feature (B) LOCATION: 885..886 . (D) OTHER INFORMATION: /note= "splice junction"
(ix) FEATURE:
(A) NAME/KEY: misc_feature (B) LOCATION: 1033..1034 (D) OTHER INFORMATION: /note= "splice junction"
(ix) FEATURE:
(A) NAME/KEY: misc_feature (B) LOCATION: 1177..1178 (D) OTHER INFORMATION: /note= "splice junction"
(ix) FEATURE:
(A) NAME/KEY: misc-feature (B) LOCATION: 1350..1351 (D) OTHER INFORMATION: /note= "splice junction"
(xi) SEQUENCE DESCRIPTION: SEQ ID N0:1:

Leu Leu Leu Thr Leu Leu Val Ser Thr Met Leu Thr Val Ser Tyr Thr TCT TCT GAT TTT CTT TCA GTG GAT GGC TGC ATT GAC TGG TCA GTG GAT" 96 Ser Ser Asp Phe Leu Ser Val Asp Gly Cys Ile Asp Trp Ser Val Asp 3 z CTCAAGACATAC ATGGCTTTG GCAGGTGAA CCAGTCCGA GTGAAA ~TGT 144 LeuLysThrTyr MetAlaLeu AlaGlyGlu ProValArg ValLys Cys AlaLeuPheTyr SerTyrIle ArgThrAsn TyrSerThr AlaGln Ser 50 55 60 . ..

ThrGlyLeuArg LeuMetTrp TyrLysAsn LysGlyAsp LeuGlu Glu ProIleIlePhe SerGluVal ArgMetSer LysGluGlu AspSer Ile TrpPheHisSer AlaGluAla GlnAspSer GlyPheTyr ThrCys..Val LeuArgAsnSer ThrTyrCys MetLysVal SerMetSer LeuThr Val AlaGluAsnGlu.SerGlyLeu CysTyrAsn SerArgIle ArgTyr Leu GluLysSerGlu ValThrLys ArgLysGlu IleSerCys ProAsp Met AspAspPheLys LysSerAsp GlnGluPro AspValVal TrpTyr Lys GluCysLysPro LysMetTrp ArgSerIle IleIleGln LysGly Asn t AlaLeuLeuIle GlnGluVal GlnGluGlu AspGlyGly AsnTyr Thr CysGluLeuLys TyrGluGly LysLeuVal ArgArgThr ThrGlu Leu LysValThrAla LeuLeuThr AspLysPro ProLysPro LeuPhe Pro MetGluAsnGln ProSerVal IleAspVal GlnLeuGly LysPro Leu AsnIleProCys LysAlaPhe PheGlyPhe SerGlyGlu SerGly Pro MetIleTyrTrp MetLysGly GluLysPhe IleGluGlu LeuAla Gly AGA AGG AAA GAA
GAA
GGT
GAA

HisIle ArgGluGly Ile LeuLeuLys GluHis LeuGlyGlu Glu Arg GCA ATC

LysGlu ValGluLeu Leu PheAspSer ValVal GluAlaAsp Ala Ile 305 310 315 . .320 TGC GTT

LeuAla AsnTyrThr His GluAsnArg AsnGly ArgLysHis Cys Val CGT AAG

AlaSer ValLeuLeu Lys AspLeuIle TyrLys IleGluLeu Arg Lys GCA TTC

AlaGly GlyLeuGly Ile LeuLeuLeu ValLeu LeuValVal Ala Phe AAC GAA

IleTyr LysCysTyr Ile LeuMetLeu PheTyr ArgGlnHis Asn Glu ACT GAT

PheGly AlaAspGlu.ThrAsn AspAsnLys GluTyr AspAlaTyr Asp GTG CAA

LeuSer TyrThrLys Asp AspThrLeu AspCys AspAsnPro Val Gln GCT GAA

GluGlu GluGlnPhe Leu ValLeuPro AspVal LeuGluLys Ala Glu CTC ATC

HisTyr GlyTyrLys Phe ProGluArg AspLeu IleProSer Leu Ile GAA CTC

GlySer AlaTyrMet Asp ThrArgTyr ValGlu GlnSerArg Glu Leu CTA CCA

ArgLeu IleIleVal Thr AspTyrIle LeuArg ArgGlyTrp Leu Pro GAA AGA

SerIle PheGluLeu Ser LeuHisAsn MetLeu ValSerGly Glu Arg TTG GAG

GluIle LysValIle Ile CysThrGlu LeuLys GlyLysVal Leu Glu GAA CTA

AsnCys GlnGluVal Ser LysArgSer IleLys LeuLeuSer Glu Leu GGA AAA

LeuIle LysTrpLys Ser SerSerLys LeuAsn SerLysPhe Gly Lys Trp Lys His Leu Val Tyr Glu Met Pro Ile Lys Lys Lys Glu Met Leu Pro Arg Cys His Val Leu Asp Ser Ala Glu Gln Gly Leu Phe Gly Glu 565 570 5a5 ..

Leu Gln Pro (2) INFORMATION FOR SEQ ID N0:2:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 579 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID N0:2:
Leu Leu Leu Thr Leu Leu Val Ser Thr Met Leu Thr Val Ser Tyr Thr Ser Ser Asp Phe Leu Ser Val Asp Gly Cys Ile Asp Trp Ser Val Asp Leu Lys Thr Tyr Met Ala Leu Ala Gly Glu Pro Val Arg Val Lys Cys Ala Leu Phe Tyr Ser Tyr Ile Arg Thr Asn Tyr Ser Thr Ala Gln Ser Thr Gly Leu Arg Leu Met Trp Tyr Lys Asn Lys Giy Asp Leu Glu Glu Pro Ile Ile Phe Ser Glu Val Arg Met Ser Lys Glu Glu Asp Ser Ile Trp Phe His Ser Ala Glu Ala Gln Asp Ser Gly Phe Tyr Thr Cys Val Leu Arg Asn Ser Thr Tyr Cys Met Lys Val Ser Met Ser Leu Thr Val Ala Glu Asn Glu Ser Gly Leu Cys Tyr Asn Ser Arg Ile Arg Tyr Leu Glu Lys Ser Glu Val Thr Lys Arg Lys Glu Ile Ser Cys Pro Asp Met Asp Asp Phe Lys Lys Ser Asp Gln Glu Pro Asp Val Val Trp Tyr Lys Glu Cys Lys Pro Lys Met Trp Arg Ser Ile Ile Ile Gln Lys Gly Asn Ala Leu Leu Ile Gln Glu Val Gln Glu Glu Asp Gly Gly Asn Tyr Thr Cys Glu Leu Lys Tyr Glu Gly Lys Leu Val Arg Arg Thr Thr Glu Leu Lys Val Thr Ala Leu Leu Thr Asp Lys Pro Pro Lys Pro Leu Phe Pro Met Glu Asn Gln Pro Ser Val Ile Asp Val Gln Leu Gly Lys Pro.l,eu Asn Ile Pro Cys Lys Ala Phe Phe Gly Phe Ser Gly Glu Ser Gly Pro Met Ile Tyr Trp Met Lys Gly Glu Lys Phe Ile Glu Glu Leu Ala Gly His Ile Arg Glu Gly Glu Ile Arg Leu Leu Lys Glu His Leu Gly Glu Lys Glu Val Glu Leu Ala Leu Ile Phe Asp Ser Val Val Glu Ala Asp Leu Ala Asn Tyr Thr Cys His Val Glu Asn Arg Asn Gly Arg Lys His Ala Ser Val Leu Leu Arg Lys Lys Asp Leu Ile Tyr Lys Ile Glu Leu Ala Gly Gly Leu Gly Ala Ile Phe Leu Leu Leu Val Leu Leu Val Val Ile Tyr Lys Cys Tyr Asn Ile Glu Leu Met Leu Phe Tyr Arg Gln His Phe Gly Ala Asp Glu Thr Asn Asp Asp Asn Lys Glu Tyr Asp Ala Tyr Leu Ser Tyr Thr Lys Val Asp Gln Asp Thr Leu Asp Cys Asp Asn Pro Glu Glu Glu Gln Phe Ala Leu Glu Val Leu Pro Asp Val Leu Glu Lys His Tyr Gly Tyr Lys Leu Phe Ile Pro Glu Arg Asp Leu Ile Pro Ser Gly Ser Ala Tyr Met Glu Asp Leu Thr Arg Tyr Val Glu Gln Ser Arg Arg Leu Ile Ile Val Leu Thr Pro Asp Tyr Ile Leu Arg Arg Gly Trp Ser Ile Phe Glu Leu Glu Ser Arg Leu His Asn Met Leu Val Ser Gly Glu Ile Lys Val Ile Leu Ile Glu Cys Thr Glu Leu Lys Gly Lys Val Asn Cys Gln Glu Val Glu Ser Leu Lys Arg Ser Ile Lys Leu Leu Ser Leu Ile Lys Trp Lys Gly Ser Lys Ser Ser Lys Leu Asn Ser Lys Phe Trp Lys His Leu Val Tyr Glu Met Pro Ile Lys Lys Lys Glu Met Leu Pro Arg Cys His Val Leu Asp Ser Ala Glu Gln Gly Leu Phe Gly Glu Leu Gln Pro . ..
(2) INFORMATION FOR SEQ ID N0:3:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 2061 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single , (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA
(ix) FEATURE:
(A) NAME/KEY: CDS
(B) LOCATION: 1..2058 (xi)SEQUENCE ID N0:3:
DESCRIPTION:
SEQ

TTT GCC GTC GTC

MetLys Pro Pro LeuLeu LeuVal CysSer Val Ser Phe Ala Val Val ATG AAG TCT TGC

ThrAsn Leu Lys ValSer ArgAsn ValAsp Gly Ile Met Lys Ser Cys GAT ACA GCT GAA

AspTrp Ser Val LeuLys TyrMet LeuAla Gly Pro Asp Thr Ala Glu TGT TTC TAT AAC

ValArg Val Lys AlaLeu TyrSer IleArg Thr Tyr Cys Phe Tyr Asn AGC CTC ATG AAC

SerThr Ala Gln ThrGly ArgLeu TrpTyr Lys Lys Ser Leu Met Asn GAG ATC GAG AGC

GlyAsp Leu Glu ProIle PheSer ValArg Met Lys Glu Ile Glu Ser ATA CAC GAG AGT

GluGlu Asp Ser TrpPhe SerAla AlaGln Asp Gly Ile His Glu Ser GTT AAC TAT GTG

PheTyr Thr Cys LeuArg SerThr CysMet Lys Ser Val Asn Tyr Val GTT AAT GGC AAC

MetSer Leu Thr AlaGlu GluSer LeuCys Tyr Ser Val Asn Gly Asn ATC GAA AAA
TCT GAA
GTC ACT
AAA AGA
AAG GAG
ATC

ArgIle ArgTyr Leu Lys Glu Val LysArg Lys Ile Glu Ser Thr Glu GAT TTT TCC CCT

SerCys ProAsp Met Asp Lys Lys AspGln Glu Asp Asp Phe Ser Pro GAA AAG ATG ATA

ValVal TrpTyr Lys Cys Pro Lys TrpArg Ser Ile Glu Lys Met Ile GCT CTG GAA GAA

IleGln LysGly Asn Leu Ile Gln ValGln Glu Asp Ala Leu Glu Glu TGT CTT GAA GTA

GlyGly AsnTyr Thr Glu Lys Tyr GlyLys Leu Arg Cys Leu Glu Val AAA ACA CTC CCT

ArgThr ThrGlu Leu Val Ala Leu ThrAsp Lys Pro Lys Thr Leu Pro ATG AAT AGT GTC

LysPro LeuPhe Pro Glu Gln Pro ValIle Asp Gln Met Asn Ser Val AAC CCC GCA TTC

LeuGly LysPro Leu Ile Cys Lys PhePhe Gly Ser Asn Pro Ala Phe ATG TAC AAA TTT

GlyGlu SerGly Pro Ile Trp Met GlyGlu Lys Ile Met Tyr Lys Phe CAC AGA GAA CTC

GluGlu LeuAla Gly Ile Glu Gly IleArg Leu Lys His Arg Glu Leu AAA GTT GCA GAC

GluHis LeuGly Glu Glu Glu Leu LeuIle Phe Ser Lys Val Ala Asp CTG AAT TGC AAC

ValVal GluAla Asp Ala Tyr Thr HisVal Glu Arg Leu Asn Cys Asn GCC GTT CGT TTA

AsnGly ArgLys His Ser Leu Leu LysLys Asp Ile Ala Val Arg Leu GCA GGC GCA CTC

TyrLys IleGlu Leu Gly Leu Gly IlePhe Leu Leu Ala Gly Ala Leu ATT AAA AAC ATG

ValLeu LeuVal Val Tyr Cys Tyr IleGlu Leu Leu Ile Lys Asn Met TTT GCT ACT AAC

PheTyr ArgGln His Gly Asp Glu AsnAsp Asp Lys Phe Ala Thr Asn GAT TAC ACT
ACA
AAA
GTG

GluTyrAsp AlaTyrLeu SerTyr ThrLysVal AspGlnAsp ThrLeu 405 410 415.

AspCysAsp AsnProGlu GluGlu GlnPheAla LeuGluVal LeuPro 420 425 430 , ., AspValLeu GluLysHis TyrGly TyrLysLeu ~PheIlePro GluArg AspLeuIle ProSerGly ThrTyr MetGluAsp LeuThrArg TyrVal GluGlnSer ArgArgLeu IleIle ValLeuThr ProAspTyr IleLeu ArgArgGly TrpSerIle PheGlu LeuGluSer ArgLeuHis AsnMet LeuValSer GlyGluIle LysVal IleLeuIle GluCysThr GluLeu LysGlyLys ValAsnCys GlnGlu ValGluSer LeuLysArg SerIle LysLeuLeu SerLeuIle LysTrp LysGlySer LysSerSer LysLeu AsnSerLys PheTrpLys HisLeu ValTyrGlu MetProIle LysLys LysGluMet LeuProArg CysHis ValLeuAsp SerAlaGlu GlnGly LeuPheGly GluLeuGln ProIle ProSerIle AlaMetThr SerThr SerAlaThr LeuValSer SerGln AlaAspLeu ProGluPhe HisPro SerAspSer MetGlnIle ArgHis CysCysArg GlyTyrLys HisGlu IleProAla ThrThrLeu ProVal ProSerLeu GlyAsnHis HisThr TyrCysAsn LeuProLeu ThrLeu LeuAsnGly GlnLeuPro LeuAsn Asn Thr Leu Lys Asp Thr Gln Glu Phe His Arg Asn Ser Ser Leu Leu Pro Leu Ser Ser Lys Glu Leu Ser Phe Thr Ser Asp Ile Trp (2) INFORMATION FOR SEQ ID N0:4:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 686 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID N0:4:
Met Lys Pro Pro Phe Leu Leu Ala Leu Val Val Cys Ser Val Val Ser Thr Asn Leu Lys Met Val Ser Lys Arg Asn Ser Val Asp Gly Cys Ile Asp Trp Ser Val Asp Leu Lys Thr Tyr Met Ala Leu Ala Gly Glu Pro Val Arg Val Lys Cys Ala Leu Phe Tyr Ser Tyr Ile Arg Thr Asn Tyr Ser Thr Ala Gln Ser Thr Gly Leu Arg Leu Met Trp Tyr Lys Asn Lys Gly Asp Leu Glu Glu Pro Ile Ile Phe Ser Glu Val Arg Met Ser Lys Glu Glu Asp Ser Ile Trp Phe His Ser Ala Glu Ala Gln Asp Ser Gly Phe Tyr Thr Cys Val Leu Arg Asn Ser Thr Tyr Cys Met Lys Val Ser Met Ser Leu Thr Val Ala Glu Asn Glu Ser Gly Leu Cys Tyr Asn Ser Arg Ile Arg Tyr Leu Glu Lys Ser Glu Val Thr Lys Arg Lys Glu Ile Ser Cys Pro Asp Met Asp Asp Phe Lys Lys Ser Asp Gln Glu Pro Asp Val Val Trp Tyr Lys Glu Cys Lys Pro Lys Met Trp Arg Ser Ile Ile Ile Gln Lys Gly Asn Ala Leu Leu Ile Gln Glu Val Gln Glu Glu Asp"

Gly Gly Asn Tyr Thr Cys Glu Leu Lys Tyr Glu Gly Lys Leu Val Arg Arg Thr Thr Glu Leu Lys Val Thr Ala Leu Leu Thr Asp Lys Pro Pro Lys Pro Leu Phe Pro Met Glu Asn Gln Pro Ser Val Ile Asp Val Gln Leu Gly Lys Pro Leu Asn Ile Pro Cys Lys Ala Phe Phe Gly P~e",Ser Gly Glu Ser Gly Pro Met Ile Tyr Trp Met Lys Gly Glu Lys Phe Ile Glu Glu Leu Ala Gly His Ile Arg Glu Gly Glu Ile Arg Leu Leu Lys Glu His Leu Gly Glu Lys Glu Val Glu Leu Ala Leu Ile Phe Asp Ser Val Val Glu Ala Asp Leu Ala Asn Tyr Thr Cys His Val Glu Asn Arg Asn Gly Arg Lys His Ala Ser Val Leu Leu Arg Lys Lys Asp Leu Ile Tyr Lys Ile Glu Leu Ala Gly Gly Leu Gly Ala Ile Phe Leu Leu Leu Val Leu Leu Val Va1 Ile Tyr Lys Cys Tyr Asn Ile Glu Leu Met Leu Phe Tyr Arg Gln His Phe Gly Ala Asp Glu Thr Asn Asp Asp Asn Lys Glu Tyr Asp Ala Tyr Leu Ser Tyr Thr Lys Val Asp Gln Asp Thr Leu Asp Cys Asp Asn Pro Glu Glu Glu Gln Phe Ala Leu Glu Val Leu Pro Asp Val Leu Glu Lys His Tyr Gly Tyr Lys Leu Phe Ile Pro Glu Arg Asp Leu Ile Pro Ser Gly Thr Tyr Met Glu Asp Leu Thr Arg Tyr Val Glu Gln Ser Arg Arg Leu Ile Ile Val Leu Thr Pro Asp Tyr Ile Leu Arg Arg Gly Trp Ser Ile Phe Glu Leu Glu Ser Arg Leu His Asn Met Leu Val Ser Gly Glu Ile Lys Val Ile Leu Ile Glu Cys Thr Glu Leu Lys Gly Lys Val Asn Cys Gln Glu Val Glu Ser Leu Lys Arg Ser Ile Lys Leu Leu Ser Leu Ile Lys Trp Lys Gly Ser Lys Ser Ser Lys Leu Asn Ser Lys Phe Trp Lys His Leu Val Tyr Glu Met Pro Ile Lys Lys Lys Glu Met Leu Pro Arg Cys His Val Leu Asp Ser Ala Glu Gln Gly Leu Phe Gly Glu Leu Gln Pro Ile Pro Ser Ile Ala Met Thr Ser Thr Ser Ala Thr Leu Val Ser Ser Gln Ala Asp Leu Pro Glu Phe H~s"Pro Ser Asp Ser Met Gln Ile Arg His Cys Cys Arg Gly Tyr Lys His Glu Ile Pro Ala Thr Thr Leu Pro Val Pro Ser Leu Gly Asn His His Thr Tyr Cys Asn Leu Pro Leu Thr Leu Leu Asn Gly Gln Leu Pro Leu Asn Asn Thr Leu Lys Asp Thr Gln Glu Phe His Arg Asn Ser Ser Leu Leu Pro Leu Ser Ser Lys Glu Leu Ser Phe Thr Ser Asp Ile Trp (2) INFORMATION FOR SEQ ID N0:5:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 482 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA
(ix) FEATURE:
(A) NAME/KEY: CDS
(B) LOCATION: 1..480 (ix) FEATURE:
(A) NAME/KEY: misc_feature (B) LOCATION: 9 (D) OTHER INFORMATION: /note= "residues 9, 459, 462, 469, and 474 are indicated as C; each may be A, C, G, or T "
(ix) FEATURE:
(A) NAME/KEY: misc_feature (B) LOCATION: 246 (D) OTHER INFORMATION: /note= "residue 246 indicated as C, may be C or G"
(ix) FEATURE:
(A) NAME/KEY: misc_feature (B) LOCATION: 321 (D) OTHER INFORMATION: /note= "residues 321, 335, 360, and 423 are indicated as C; each may be C or T"
(ix) FEATURE:
(A) NAME/KEY: misc feature (B) LOCATION: 426 (D) OTHER INFORMATION: /note= "residue 426 indicated as C, may be A or C"

(xi) SEQUENCE DESCRIPTION: SEQ ID N0:5:

Lys Tyr Gly Tyr Ser Leu Phe Phe Leu Glu Arg Asn Val Ala Pro Gly Gly Val Tyr Ala Glu Asp Ile Val Ser Ile Ile Lys Arg Ser Arg Arg ATA ACC GTC ATC

GlyIle PheIle Leu Pro AsnTyrVal AsnGlyPro SerIlePhe Thr GTG

GluLeu GlnAla Ala Asn LeuAlaLeu AspAspGln ThrLeuLys Val TTC

LeuIle LeuIle Lys Cys TyrPheGln GluProGlu SerLeuPro Phe GCT

HisLeu ValLys Lys Leu ArgValLeu ProThrVal ThrTrpArg Ala CCT

GlyLeu LysSer Val Pro AsnSerArg PheTrpAla LysMetArg Pro AAA

TyrHis MetPro Val Asn LeuSerGly IleHisVal GlyThrSer Lys AGG

SerArg IleThr Ser Asp PhePheSer GlyLysAsp SerValGlu Arg GGA

GlnLys ProTrp Gly Ala ProSerLeu LysGlyArg CysAsnGlu Gly (2) INFORMATION FOR SEQ ID N0:6:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 160 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID N0:6:
Lys Tyr Gly Tyr Ser Leu Phe Phe Leu Glu Arg Asn Val Ala Pro Gly Gly Val Tyr Ala Glu Asp Ile Val Ser Ile Ile Lys Arg Ser Arg Arg Gly Ile Phe Ile Leu Thr Pro Asn Tyr Val Asn Gly Pro Ser Ile Phe Glu Leu Gln Ala Ala Val Asn Leu Ala Leu Asp Asp Gln Thr Leu Lys Leu Ile Leu Ile Lys Phe Cys Tyr Phe Gln Glu Pro Glu Ser Leu Pro w His Leu Val Lys Lys Ala Leu Arg Val Leu Pro Thr Val Thr Trp Arg Gly Leu Lys Ser Val Pro Pro Asn Ser Arg Phe Trp Ala Lys Met Arg Tyr His Met Pro Val Lys Asn Leu Ser Gly Ile His Val Gly Thr Ser 115 120 125 , Ser Arg Ile Thr Ser Arg Asp Phe Phe Ser Gly Lys Asp Ser Val Glu Gln Lys Pro Trp Gly Gly Ala Pro Ser Leu Lys Gly Arg Cys Asn Glu (2) INFORMATION FOR SEQ ID N0:7:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 1404 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA
(ix) FEATURE:
(A) NAME/KEY: CDS
(B) LOCATION: 1..1401 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:7:

AGC ATT

PheProArgSer ProTyrAsp ValAlaCys CysValLys MetIleLeu GluValLysPro GlnThrAsn AlaSerCys GluTyrSer AlaSerHis LysGlnAspLeu LeuLeuGly SerThrGly SerIleSer CysProSer LeuSerCysGln SerAspAla GlnSerPro AlaValThr TrpTyrLys AsnGlyLysLeu LeuSerVal GluArgSer AsnArgIle ValValAsp GluValTyrAsp TyrHisGln GlyThrTyr ValCysAsp TyrThrGln GAT GTG
ACT AGA
GTG
AGT
TCG
TGG
ACA
GTC
AGA
GCT

Ser AspThr Val Ser Trp Thr Val Arg ValVal GlnVal Arg Ser Ala GAC GAT

Thr IleVal Gly Thr Lys Leu Lys Pro IleLeu AspPro ~lal Asp Asp GAA CCT

Glu AspThr Leu Val Glu Leu Gly Lys LeuThr IleSer Cys Glu Pro GGC AAC

Lys AlaArg Phe Phe Glu Arg Val Phe ProVal IleLys Trp Gly Asn TCT GTC

Tyr IleLys Asp Asp Leu Glu Trp Glu SerVal ProGlu Ala Ser Val TCC ATC

Lys SerIle Lys Thr Leu Lys Asp Glu IleGlu ArgAsn Ile Ser Ile GTC CGC

Ile LeuGlu Lys Thr Gln Arg Asp Leu ArgLys PheVal Cys Val Arg TCC CAG

Phe ValGln Asn Ile Gly Asn Thr Thr SerVal GlnLeu Lys Ser Gln GTG CTG

Glu LysArg Gly Val Leu Leu Tyr Ile LeuGly ThrIle Gly Val Leu GTG CTC

Thr LeuVal Ala Leu Ala Ala Ser Ala LeuTyr ArgHis Trp Val Leu CTG CAG

Ile GluIle Val Leu Tyr Arg Thr Tyr SerLys AspGln Thr Leu Gln AAG GTA

Leu GlyAsp Lys Asp Phe Asp Ala Phe SerTyr AlaLys Trp Lys Val AGT CTG

Ser SerPhe Pro Glu Ala Thr Ser Ser SerGlu GluHis Leu Ser Leu TTT AAC

Ala LeuSer Leu Pro Asp Val Leu Glu LysTyr GlyTyr Ser Phe Asn GAA GGA

Leu CysLeu Leu Arg Asp Val Ala Pro GlyVal TyrAla Glu Glu Gly ATT GAG

Asp IleVal Ser Ile Lys Arg Ser Arg ValIle PheIle Leu Ile Glu GTC GGA CTA
CCC CAA
AGT GCA
ATC GCA

SerPro AsnTyr ValAsn Pro Ser PheGluLeu Gln Ala Ala Gly Ile GAT CTG

ValAsn LeuAla LeuAsp Gln Thr LysLeuIle Leu I~e"Lys Asp Leu CCA CTA

PheCys TyrPhe GlnGlu Glu Ser ProHisLeu Val Lys Lys Pro Leu ACA TGG

AlaLeu ArgVal LeuPro Val Thr ArgGlyLeu Lys Ser Val Thr Trp TGG ATG

ProPro AsnSer ArgPhe Ala Lys ArgTyrHis Met Pro Val Trp Met ACG CAG

LysAsn SerGln GlyPhe Trp Asn LeuArgIle Thr Ser Arg Thr Gln CTC ACA

IlePhe GlnTrp LysGly Ser Arg GluThrThr Gly Glu Glu Leu Thr LeuPro Ala (2) INFORMATION FOR SEQ ID N0:8:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 467 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID N0:8:
Phe Pro Arg Ser Pro Tyr Asp Val Ala Cys Cys Val Lys Met Ile Leu Glu Val Lys Pro Gln Thr Asn Ala Ser Cys Glu Tyr Ser Ala Ser His Lys Gln Asp Leu Leu Leu Gly Ser Thr Gly Ser Ile Ser Cys Pro Ser Leu Ser Cys Gln Ser Asp Ala Gln Ser Pro Ala Val Thr Trp Tyr Lys Asn Gly Lys Leu Leu Ser Val Glu Arg Ser Asn Arg Ile Val Val Asp 65 70 75 80w Glu Val Tyr Asp Tyr His Gln Gly Thr Tyr Val Cys Asp Tyr Thr Gln Ser Asp Thr Val Ser Ser Trp Thr Val Arg Ala Val Val Gln Val Arg Thr Ile Val Gly Asp Thr Lys Leu Lys Pro Asp Ile Leu Asp Pro Val Glu Asp Thr Leu Glu Val Glu Leu Gly Lys Pro Leu Thr Ile Ser ~Cys Lys Ala Arg Phe Gly Phe Glu Arg Val Phe Asn Pro Val Ile Lys Trp Tyr Ile Lys Asp Ser Asp Leu Glu Trp Glu Val Ser Val Pro Glu Ala Lys Ser Ile Lys Ser Thr Leu Lys Asp Glu Ile Ile Glu Arg Asn Ile Ile Leu Glu Lys Val Thr Gln Arg Asp Leu Arg Arg Lys Phe Val Cys Phe Val Gln Asn Ser Ile Gly Asn Thr Thr Gln Ser Val Gln Leu Lys Glu Lys Arg Gly Val Val Leu Leu Tyr Ile Leu Leu Gly Thr Ile Gly Thr Leu Val Ala Val Leu Ala Ala Ser Ala Leu Leu Tyr Arg His Trp Ile Glu Ile Val Leu Leu Tyr Arg Thr Tyr Gln Ser Lys Asp Gln Thr Leu Gly Asp Lys Lys Asp Phe Asp Ala Phe Val Ser Tyr Ala Lys Trp Ser Ser Phe Pro Ser Glu Ala Thr Ser Ser Leu Ser Glu Glu His Leu Ala Leu Ser Leu Phe Pro Asp Val Leu Glu Asn Lys Tyr Gly Tyr Ser Leu Cys Leu Leu Glu Arg Asp Val Ala Pro Gly Gly Val Tyr Ala Glu Asp Ile Val Ser Ile Ile Lys Arg Ser Arg Glu Val Ile Phe Ile Leu Ser Pro Asn Tyr Val Asn Gly Pro Ser Ile Phe Glu Leu Gln Ala Ala Val Asn Leu Ala Leu Asp Asp Gln Thr Leu Lys Leu Ile Leu Ile Lys Phe Cys Tyr Phe Gln Glu Pro Glu Ser Leu Pro His Leu Val Lys Lys Ala Leu Arg Val Leu Pro Thr Val Thr Trp Arg Gly Leu Lys Ser Val Pro Pro Asn Ser Arg Phe Trp Ala Lys Met Arg Tyr His Met Pro Val Lys Asn Ser Gln Gly Phe Thr Trp Asn Gln Leu Arg Ile Thr Ser Arg Ile Phe Gln Trp Lys Gly Leu Ser Arg Thr Glu Thr Thr Gly Glu Glu Leu Pro Ala (2) INFORMATION FOR SEQ ID N0:9:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 2314 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA
(ix) FEATURE:
(A) NAME/KEY: CDS
(B) LOCATION: 109..1905 (xi)SEQUENCE SEQID
DESCRIPTION: N0:9:

GGAATTCGGA ACTTTAATCT
TACTCAGGGC
AGAGTTCTGA

TGAAGTTATT ATG
GGAGTGATGA CTC
CAGGAACACG TGT

Me t Leu Cys TTT

Leu GlyTrpIle LeuTrp LeuValAla GlyGluArg IleLysGly Phe GGT

Phe AsnIleSer CysSer ThrLysLys LeuLeuTrp ThrTyrSer Gly GAG

Thr ArgSerGlu GluPhe ValLeuPhe CysAspLeu ProGluPro Glu TTC

Gln LysSerHis CysHis ArgAsnArg LeuSerPro LysGlnVal Phe CCC

Pro GluHisLeu PheMet GlySerAsn AspLeuSer AspValGln Pro CCT

Trp TyrGlnGln SerAsn GlyAspPro LeuGluAsp IleArgLys Pro ATC

Ser TyrProHis IleGln AspLysCys ThrLeuHis PheLeuThr Ile AAT

Pro GlyValAsn SerGly SerTyrIle CysArgPro LysMetIle Asn CCC GTC AAG ATG ATT GAA
GTT

Lys Ser Tyr Asp Val Ala Cys Cys Met LeuGlu Val Pro Val Lys Ile CAG TAT TCC TCA

Lys Pro Thr Asn Ala Ser Cys Glu Ala HisLys"~Gln Gln Tyr Ser Ser CTT ATT TCT CCC

Asp Leu Leu Gly Ser Thr Gly Ser Cys SerLeu Ser Leu Ile Ser Pro AGT GTA ACC TAC

Cys Gln Asp Ala Gln Ser Pro Ala Trp LysAsn Gty Ser Val Thr Tyr CTC CGA ATC GTG

Lys Leu Ser Val Glu Arg Ser Asn Val AspGlu Val Leu Arg Ile Val TAT TGT GAT ACT

Tyr Asp His Gln Gly Thr Tyr Val Tyr GlnSer Asp Tyr Cys Asp Thr AGT GTT GTT GTG

Thr Val Ser Trp Thr Val Arg Ala Gln ArgThr Ile Ser Val Val Val GAC ATT CTG CCT

Val Gly Thr Lys Leu Lys Pro Asp Asp ValGlu Asp Asp Ile Leu Pro GAA TTA ACT AGC

Thr Leu Val Glu Leu Gly Lys Pro Ile CysLys Ala Glu Leu Thr Ser GGC CCT GTC AAA

Arg Phe Phe Glu Arg Val Phe Asn Ile TrpTyr Ile Gly Pro Val Lys TCT TCA GTA GAG

Lys Asp Asp Leu Glu Trp Glu Val Pro AlaLys Ser Ser Ser Val Glu TCC ATT GAG AAT

Ile Lys Thr Leu Lys Asp Glu Ile Arg IleIle Leu Ser Ile Glu Asn GTC AGG AAG GTT

Glu Lys Thr Gln Arg Asp Leu Arg Phe CysPhe Val Val Arg Lys Val TCC TCC GTC CTG

Gln Asn Ile Gly Asn Thr Thr Gln Gln LysGlu Lys Ser Ser Val Leu GTG CTT GGC ATC

Arg Gly Val Leu Leu Tyr Ile Leu Thr GlyThr Leu Val Leu Gly Ile GTG CTC TAC CAC

Val Ala Leu Ala Ala Ser Ala Leu Arg TrpIle Glu Val Leu Tyr His 20 _ GTG AGC ACG GGG
CTG
CTG
TAC
CGG
ACC

Ile ValLeu Leu Tyr Arg Tyr Gln LysAsp Gln LeuGly Thr Ser Thr GCT TCC TGG

Asp LysLys Asp Phe Asp Phe Val TyrAla Lys Ser.,aSer Ala Ser Trp TCA AGT TTG

Phe ProSer Glu Ala Thr Ser Leu GluGlu His AlaLeu Ser Ser Leu TTA AAA AGC

Ser LeuPhe Pro Asp Val Glu Asn TyrGly Tyr LeuCys Leu Lys Ser GCT GGA GAA

Leu LeuGlu Arg Asp Val Pro Gly ValTyr Ala AspIle Ala Gly Glu AGC GGA TTG

Val SerIle Ile Lys Arg Arg Arg IlePhe Ile SerPro Ser Gly Leu AGT GAA GCA

Asn TyrVal Asn Gly Pro Ile Phe LeuGln Ala ValAsn Ser Glu Ala ACA CTC AAG

Leu AlaLeu Asp Asp Gln Leu Lys IleLeu Ile PheCys Thr Leu Lys TCT CAT AAA

Tyr PheGln Glu Pro Glu Leu Pro LeuVal Lys AlaLeu Ser His Lys ACT GGC GTT

Arg ValLeu Pro Thr Val Trp Arg LeuLys Ser ProPro Thr Gly Val AAA TAC GTG

Asn SerArg Phe Trp A1a Met Arg HisMet Pro LysAsn Lys Tyr Val AAC AGA AGG

Ser GlnGly Phe Thr Trp Gln Leu IleThr Ser IlePhe Asn Arg Arg AGA ACA GAA ACC AGC

Gln Lys Gly Leu Ser Thr Glu ThrGly SerGln Trp Arg Thr Arg Ser AAG CCTCCAGTCC
AGTCCCTGGG

Pro Lys Glu Trp 21 _ (2) INFORMATION FOR SEQ ID NO:10:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 599 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO:10:
Met Leu Cys Leu Gly Trp Ile Phe Leu Trp Leu Val Ala Gly Glu Arg Ile Lys Gly Phe Asn Ile Ser Gly Cys Ser.Thr Lys Lys Leu Leu Trp Thr Tyr Ser Thr Arg Ser Glu Glu Glu Phe Val Leu Phe Cys Asp Leu Pro Glu Pro Gln Lys Ser His Phe Cys His Arg Asn Arg Leu Ser Pro Lys Gln Val Pro Glu His Leu Pro Phe Met Gly Ser Asn Asp Leu Ser Asp Val Gln Trp Tyr Gln Gln Pro Ser Asn Gly Asp Pro Leu Glu Asp Ile Arg Lys Ser Tyr Pro His Ile Ile Gln Asp Lys Cys Thr Leu His Phe Leu Thr Pro Gly Val Asn Asn Ser Gly Ser Tyr Ile Cys Arg Pro Lys Met Ile Lys Ser Pro Tyr Asp Val Ala Cys Cys Val Lys Met Ile Leu Glu Val Lys Pro Gln Thr Asn Ala Ser Cys Glu Tyr Ser Ala Ser His Lys Gln Asp Leu Leu Leu Gly Ser Thr Gly Ser Ile Ser Cys Pro Ser Leu Ser Cys Gln Ser Asp Ala Gln Ser Pro Ala Val Thr Trp Tyr Lys Asn Gly Lys Leu Leu Ser Val Glu Arg Ser Asn Arg Ile Val Val Asp Glu Val Tyr Asp Tyr His Gln Gly Thr Tyr Val Cys Asp Tyr Thrw Gln Ser Asp Thr Val Ser Ser Trp Thr Val Arg AIa Val Val Gln Val Arg Thr Ile Val Gly Asp Thr Lys Leu Lys Pro Asp Ile Leu Asp Pro Val Glu Asp Thr Leu Glu Val Glu Leu Gly Lys Pro Leu Thr Ile Ser Cys Lys Ala Arg Phe Gly Phe Glu Arg Val Phe Asn Pro Val Ile ~ys Trp Tyr Ile Lys Asp Ser Asp Leu Glu Trp Glu Val Ser Val Pro Glu Ala Lys Ser Ile Lys Ser Thr Leu Lys Asp Glu Ile Ile Glu Arg Asn Ile Ile Leu Glu Lys Val Thr Gln Arg Asp Leu Arg Arg Lys Phe Val Cys Phe Val Gln Asn Ser Ile Gly Asn Thr Thr Gln Ser Val Gln Leu Lys Glu Lys Arg Gly Val Val Leu Leu Tyr Ile Leu Leu Gly Thr Ile Gly Thr Leu Val Ala Val Leu Ala Ala Ser Ala Leu Leu Tyr Arg His Trp Ile Glu Ile Val Leu Leu Tyr Arg Thr Tyr Gln Ser Lys Asp Gln Thr Leu Gly Asp Lys Lys Asp Phe Asp Ala Phe Val Ser Tyr Ala Lys Trp Ser Ser Phe Pro Ser Glu Ala Thr Ser Ser Leu Ser Glu Glu His Leu Ala Leu Ser Leu Phe Pro Asp Val Leu Glu Asn Lys Tyr Gly Tyr Ser Leu Cys Leu Leu Glu Arg Asp Val Ala Pro Gly Gly Val Tyr Ala Glu Asp Ile Val Ser Ile Ile Lys Arg Ser Arg Arg Gly Ile Phe Ile Leu Ser Pro Asn Tyr Val Asn Gly Pro Ser Ile Phe Glu Leu Gln Ala Ala Val Asn Leu Ala Leu Asp Asp Gln Thr Leu Lys Leu Ile Leu Ile Lys Phe Cys Tyr Phe Gln Glu Pro Glu Ser Leu Pro His Leu Val Lys Lys Ala Leu Arg Val Leu Pro Thr Val Thr Trp Arg Gly Leu Lys Ser Val Pro Pro Asn Ser Arg Phe Trp Ala Lys Met Arg Tyr His Met Pro 545 550 555 560.
Val Lys Asn Ser Gln Gly Phe Thr Trp Asn Gln Leu Arg Ile Thr Ser Arg Ile Phe Gln Trp Lys Gly Leu Ser Arg Thr Glu Thr Thr Gly Arg Ser Ser Gln Pro Lys Glu Trp (2) INFORMATION FOR SEQ ID NO:11:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 768 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA
(ix) FEATURE:
(A) NAME/KEY: CDS
(B) LOCATION: 1..360 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:11:

Ala Ala Val Asn Leu Ala Leu Val Asp Gln Thr Leu Lys Leu Ile Leu Ile Lys Phe Cys Ser Phe Gln Glu Pro Glu Ser Leu Pro Tyr Leu Val Lys Lys Ala Leu Arg Val Leu Pro Thr Val Thr Trp Lys Gly Leu Lys Ser Val His Ala Ser Ser Arg Phe Trp Thr Gln Ile Arg Tyr His Met Pro Val Lys Asn Ser Asn Arg Phe Met Phe Asn Gly Leu Arg Ile Phe Leu Lys Gly Phe Ser Pro Glu Lys Asp Leu Val Thr Gln Lys Pro Leu Glu Gly Met Pro Lys Ser Gly Asn Asp His Gly Ala Gln Asn Leu Leu Leu Tyr Ser Asp Gln Lys Arg Cys (2) INFORMATION FOR SEQ ID N0:12:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 120 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID N0:12:
Ala Ala Val Asn Leu Ala Leu Val Asp Gln Thr Leu Lys Leu Ile Leu Ile Lys Phe Cys Ser Phe Gln Glu Pro Glu Ser Leu Pro Tyr Leu Val Lys Lys Ala Leu Arg Val Leu Pro Thr Val Thr Trp Lys Gly Leu Lys Ser Val His Ala Ser Ser Arg Phe Trp Thr Gln Ile Arg Tyr His Met Pro Val Lys Asn Ser Asn Arg Phe Met Phe Asn Gly Leu Arg Ile Phe Leu Lys Gly Phe Ser Pro Glu Lys Asp Leu Val Thr Gln Lys Pro Leu Glu Gly Met Pro Lys Ser Gly Asn Asp His Gly Ala Gln Asn Leu Leu Leu Tyr Ser Asp Gln Lys Arg Cys (2) INFORMATION FOR SEQ ID N0:13:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 1833 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA
(ix) FEATURE:
(A) NAME/KEY: CDS
(B) LOCATION: 1..1830 (ix) FEATURE:
(A) NAME/KEY: mat~eptide (B) LOCATION: 52..1830 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:13:

TGT GCA GGA GAG AAG ACC ACA

MetSer Val Trp Leu Val Phe Leu Val GlyGlu Lys Thr Thr Cys Ala AAA AAT

GlyPhe Asn His Ser Ala Cys Ala Thr SerVal Asp Ile Phe Lys Asn 1 5 10 . ., 15 TTT GTG

AlaArg Gly Ala Glu Asn Phe Val Tyr Thr Phe Val Tyr Lys Ser Phe AAC TGT

ArgSer Lys Asn Ser Pro Met Gln Val HisGln His Lys Val Asn Cys AAG GAC

CysSer Gln Thr Cys Ser Gly Ser Gln LeuSer Asp Val Gln Lys Asp CCA CTA

TrpTyr Met Gln Pro Arg Ser Gly Ser GluGlu Ile Ser Arg Pro Leu ATG CTG

AsnSer Pro His Met Gln Ser Glu Gly HisIle Leu Ala Pro Met Leu TGT AGA

GlnThr Asn Ser Ile Trp Ser Tyr Ile ProArg Ile Arg Ser Cys Arg ACA GTC

ProGln Asp Met Ala Cys Cys Ile Lys LeuGlu Val Lys Pro Thr Val GCA CAA

GlnArg Asn Val Ser Cys Gly Asn Thr AspGlu Gln Val Leu Ala Gln TGT CCC

LeuLeu Gly Ser Thr Gly Ser Ile His SerLeu Ser Cys Gln Cys Pro TGG TAC

SerAsp Val Gln Ser Pro Glu Met Thr LysAsp Gly Arg Leu Trp Tyr GAG ATG

LeuPro Glu His Lys Lys Asn Pro Ile AlaAsp Ile Tyr Val Glu Met TAC ACA

PheAsn Gln Gly Leu Tyr Val Cys Asp GlnSer Asp Asn Val Tyr Thr AAA GTG

SerSer Trp Thr Val Arg Ala Val Val ArgThr Ile Gly Lys Lys Val GAT CCC

AspIle Asn Val Lys Pro Glu Ile Leu IleThr Asp Thr Leu Asp Pro CTC CCC AGA
GTA
CAG
TTT

Asp Val Glu Leu Gly Lys Pro Leu Thr Cys Val Gln Phe Leu Pro Arg ATA AAG TAT

Gly Phe Gln Arg Leu Ser Lys Pro Val Trp Val Lys Glu Ile Lys Tyr TTT GAG AAA

Ser Thr Gln Glu Trp Glu Met Ser Val ~Glu Arg Ile Gln Phe Glu Lys CGT ACC TTC

Ser Thr Phe Lys Asn Glu Val Ile Glu Ile Leu Arg Glu Arg Thr Phe TTT GTT TTT

Val Thr Gln Arg Asp Leu Ser Arg Lys Cys Ala Gln Asn Phe Val Phe ' CGG CTG AAG

Ser Ile Gly Asn Thr Thr Arg Thr Ile Arg Lys Glu Glu Arg Leu Lys ACG GCC ATG

Val Val Phe Val. Tyr Ile Leu Leu Gly Leu Leu Val Gly Thr Ala Met TGG TAC ATT

Val Leu Val Ala Ala Ala Phe Leu Tyr Trp Glu Val Val Trp Tyr Ile GAT GAG CTG

Leu Leu Cys Arg Thr Tyr Lys Asn Lys Thr Gly Asp Lys Asp Glu Leu TCG AAT AGC

Lys Glu Phe Asp Ala Phe Val Ser Tyr Trp Ser Pro Glu Ser Asn Ser GAA CAC GCT

Thr Asp Ala Val Gly Ser Leu Ser Glu Leu Leu Asn Leu Glu His Ala GGG TAC TTG

Phe Pro Glu Val Leu Glu Asp Thr Tyr Arg Cys Leu Leu Gly Tyr Leu TAT GCA GAC

Asp Arg Asp Val Thr Pro Gly Gly Val Asp Ile Val Ser Tyr Ala Asp TTT ATC AGT

Ile Ile Lys Lys Ser Arg Arg Gly Ile Leu Pro Ser Tyr Phe Ile Ser 450 455. 460 CAA GCA GTG

Leu Asn Gly Pro Arg Val Phe Glu Leu Ala Asn Leu Ala Gln Ala Val TTA ATT TTC

Leu Val Asp Gln Thr Leu Lys Leu Ile Lys Cys Ser Phe Leu Ile Phe GAG GAA CTT TAC GTC
AAA
AAG

Gln Pro GluSer Pro LeuVal Lys AlaLeu Arg Val Glu Leu Tyr Lys CCC TGG GGC GTC

Leu Thr ValThr Lys LeuLys Ser HisAla Ser Ser Pro Trp Gly Val 515 520 525 . ..

TTC ATT TAC GTG

Arg Trp ThrGln Arg HisMet Pro LysAsn Ser Asn Phe Ile Tyr Val TTT GGG AGA AAG

Arg Met PheAsn Leu IlePhe Leu GlyPhe Ser Pro Phe Gly Arg Lys AAG ACA AAA GGA

Glu Asp LeuVal Gln ProLeu Glu MetPro Lys Ser Lys Thr Lys Gly 560 565 570 ~ 575 AAT GCT AAC TAC

Gly Asp HisGly Gln LeuLeu Leu SerAsp Gln Lys Asn Ala Asn Tyr TGC

Arg Cys (2) INFORMATION FOR SEQ ID N0:14:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 610 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID N0:14:
Met Ser Val Trp Leu Val Phe Leu Val Cys Ala Gly Glu Lys Thr Thr Gly Phe Asn His Ser Ala Cys Ala Thr Lys Asn Ser Val Asp Ile Phe Ala Arg Gly Ala Glu Asn Phe Val Tyr Phe Val Thr Tyr Lys Ser Phe Arg Ser Lys Asn Ser Pro Met Gln Val Asn Cys His Gln His Lys Val Cys Ser Gln Thr Cys Ser Gly Ser Gln Lys Asp Leu Ser Asp Val Gln Trp Tyr Met Gln Pro Arg Ser Gly Ser Pro Leu Glu Glu Ile Ser Arg Asn Ser Pro His Met Gln Ser Glu Gly Met Leu His Ile Leu Ala Pro"

Gln Thr Asn Ser Ile Trp Ser Tyr Ile Cys Arg Pro Arg Ile Arg Ser ' 100 105 110 WO 99/19480 PCT/US98l10939 Pro Gln Asp Met Ala Cys Cys Ile Lys Thr Val Leu Glu Val Lys Pro Gln Arg Asn Val Ser Cys Gly Asn Thr Ala Gln Asp Glu Gln Val Leu Leu Leu Gly Ser Thr Gly Ser Ile His Cys Pro Ser Leu Ser Cys.,Gln Ser Asp Val Gln Ser Pro Glu Met Thr Trp Tyr Lys Asp Gly Arg Leu Leu Pro Glu His Lys Lys Asn Pro Ile Glu Met Ala Asp Ile Tyr Val Phe Asn Gln Gly Leu Tyr Val Cys Asp Tyr Thr Gln Ser Asp Asn Val Ser Ser Trp Thr Val Arg Ala Val Val Lys Val Arg Thr Ile Gly Lys 210 215 ,22 0 Asp Ile Asn Val Lys Pro Glu Ile Leu Asp Pro Ile Thr Asp Thr Leu Asp Val Glu Leu Gly Lys Pro Leu Thr Leu Pro Cys Arg Val Gln Phe Gly Phe Gln Arg Leu Ser Lys Pro Val Ile Lys Trp Tyr Val Lys Glu Ser Thr Gln Glu Trp Glu Met Ser Val Phe Glu Glu Lys Arg Ile Gln Ser Thr Phe Lys Asn Glu Val Ile Glu Arg Thr Ile Phe Leu Arg Glu Val Thr Gln Arg Asp Leu Ser Arg Lys Phe Val Cys Phe Ala Gln Asn Ser Ile Gly Asn Thr Thr Arg Thr Ile Arg Leu Arg Lys Lys Glu Glu Val Val Phe Val Tyr Ile Leu Leu Gly Thr Ala Leu Met Leu Val Gly Val Leu Val Ala Ala Ala Phe Leu Tyr Trp Tyr Trp Ile Glu Val Val Leu Leu Cys Arg Thr Tyr Lys Asn Lys Asp Glu Thr Leu Gly Asp Lys Lys Glu Phe Asp Ala Phe Val Ser Tyr Ser Asn Trp Ser Ser Pro Glu Thr Asp Ala Val Gly Ser Leu Ser Glu Glu His Leu Ala Leu Asn Leu Phe Pro Glu Val Leu Glu Asp Thr Tyr Gly Tyr Arg Leu Cys Leu Leu Asp Arg Asp Val Thr Pro Gly Gly Val Tyr Ala Asp Asp Ile Val Ser Ile Ile Lys Lys Ser Arg Arg Gly Ile Phe Ile Leu Ser Pro Ser Tyr Leu Asn Gly Pro Arg Val Phe Glu Leu Gln Ala Ala Val Asn Leu Ala Leu Val Asp Gln Thr Leu Lys Leu Ile Leu Ile Lys Phe Cys Ser Phe 480 485 490 ~ 495 Gln Glu Pro Glu Ser Leu Pro Tyr Leu Val Lys Lys Ala Leu Arg Val Leu Pro Thr Val Thr Trp Lys Gly Leu Lys Ser Val His Ala Ser Ser Arg Phe Trp Thr Gln Ile Arg Tyr His Met Pro Val Lys Asn Ser Asn Arg Phe Met Phe Asn Gly Leu Arg Ile Phe Leu Lys Gly Phe Ser Pro Glu Lys Asp Leu Val Thr Gln Lys Pro Leu Glu Gly Met Pro Lys Ser Gly Asn Asp His Gly Ala Gln Asn Leu Leu Leu Tyr Ser Asp Gln Lys Arg Cys (2) INFORMATION FOR SEQ ID N0:15:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 2259 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA
(ix) FEATURE:
(A) NAME/KEY: CDS
(H) LOCATION: 22..1863 (xi) SEQUENCE
DESCRIPTION:
SEQ ID
N0:15:

AAAGGGGAAC GGC GTG
C ATG
CTC

Met Leu Cys Leu Trp PheLeuTrp Gly Val GCA ACA AAT TCA

Phe Val Gly Glu Lys Thr Gly Phe His AlaCysAla Ala Thr Asn Ser AAA TAT AGG GCA

Thr Lys Leu Leu Trp Thr Ser Ala Gly GluAsnPhe Lys Tyr Arg Ala TTT GAG GAG AAA

Val Leu Cys Asp Leu Gln Leu Gln Gln PheSerHis Phe Glu Glu Lys AGT CCA CCT
ACA
CAA
AGT
CCT
GCT

AlaSer GlnLeuSer Pro Thr Gln Pro HisLysPro Cys Ser Ser Ala GTC TGG

GlySer GlnLysAsp Leu Ser Asp Gln TyrMetGln Pro Arg Val Trp 75 80 85 , ~, 90 AGT AAC

SerGly SerProLeu Glu Glu Ile Arg SerProHis Met Gln Ser Asn GCC CAG

SerGlu GlyMetLeu His Ile Leu Pro ThrAsnSer Ile Trp Ala Gln AGG CCC

SerTyr IleCysArg Pro Arg Ile Ser GlnAspMet Ala Cys Arg Pro 125 130 1'35 ATC GTC GTT AGA

Cys IleLysThr ValLeu Glu LysPro GlnArgAsn ValSerCys Val CAA

Gly AsnThrAla GlnAsp Glu ValLeu LeuLeuGly SerThrGly Gln AGC

Ser IleHisCys ProSer Leu CysGln SerAspVal GlnSerPro Ser GGA

Glu MetThrTrp TyrLys Asp ArgLeu LeuProGlu HisLysLys Gly ATT

Asn ProIleGlu MetAla Asp TyrVal PheAsnGln GlyLeuTyr Ile GAT

Val CysAspTyr ThrGln Ser AsnVal SerSerTrp ThrValArg Asp ATT

Ala ValValLys ValArg Thr GlyLys AspIleAsn ValLysPro Ile GAT

Glu IleLeuAsp ProIle Thr ThrLeu AspValGlu LeuGlyLys Asp GTA

Pro LeuThrLeu ProCys Arg GlnPhe GlyPheGln ArgLeuSer Val GTC

Lys ProValIle LysTrp Tyr LysGlu SerThrGln GluTrpGlu Val AGA

Met SerValPhe GluGlu Lys IleGln SerThrPhe LysAsnGlu Arg CGT GTT ACC GAT
ACC
ATC
TTC

ValIle GluArgThr Ile Leu Arg Glu ThrGln Arg Leu Phe Val Asp TTT TCC ACA

SerArg LysPheVal Cys Ala Gln Asn IleGly Asn Thr Phe Ser Thr ..

AAG GTG TAC

ArgThr IleArgLeu Arg Lys Glu Glu ValPhe Val Ile Lys Val Tyr ATG GTT GCT

LeuLeu GlyThrAla Leu Leu Val Gly LeuVal Ala Ala Met Val Ala 365 370 375 , ATT CTG ACC

PheLeu TyrTrpTyr Trp Glu Val Val LeuCys Arg Tyr Ile Leu Thr CTG AAG GCA

LysAsn LysAspGlu Thr Gly Asp Lys GluPhe Asp Phe Leu Lys Ala AGC ACT GGA

ValSer TyrSerAsn Trp Ser Pro Glu AspAla Val Ser Ser Thr Gly GCT TTC CTA

LeuSer GluGluHis Leu Leu Asn Leu ProGlu Val Glu Ala Phe Leu GGG AGA GAC GAT
TTG

AspThr TyrGlyTyr LeuCys LeuLeuAsp ArgAspVal ThrPro Arg GAT

GlyGly ValTyrAla AspIle ValSerIle IleLysLys SerArg Asp CTG

ArgGly IlePheIle SerPro SerTyrLeu AsnGlyPro ArgVal Leu GCA

PheGlu LeuGlnAla ValAsn LeuAlaLeu ValAspGln ThrLeu Ala AAG

LysLeu IleLeuIle PheCys SerPheGln GluProGlu SerLeu Lys AAG

ProTyr LeuValLys AlaLeu ArgValLeu ProThrVal ThrTrp Lys GTC

LysGly LeuLysSer HisAla SerSerArg PheTrpThr GlnIle Val GTG

ArgTyr HisMetPro LysAsn SerAsnArg PheMetPhe AsnGly Val CTC AGA ATT TTC CTG AAG 'M'T CCT GAA AAG CTA GTG ACA 1779 GGC TCC GAC

Leu Arg Ile Phe Leu Lys Phe Pro Glu Lys Leu Val Thr Gly Ser Asp ATG AAG GAC

Gln Lys Pro Leu Glu Gly Pro Ser Gly Asn His Gly Ala Met Lys Asp w Gln Asn Leu Leu Leu Tyr Ser Asp Gln Lys Arg Cys CTGTAAAGAA AAAAAP.AAAA AAAAAA 2259 (2) INFORMATION FOR SEQ ID N0:16:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 614 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID N0:16:
Met Leu Cys Leu Gly Trp Val Phe Leu Trp Phe Val Ala Gly Glu Lys Thr Thr Gly Phe Asn His Ser Ala Cys Ala Thr Lys Lys Leu Leu Trp Thr Tyr Ser Ala Arg Gly Ala Glu Asn Phe Val Leu Phe Cys Asp Leu Gln Glu Leu Gln Glu Gln Lys Phe Ser His Ala Ser Gln Leu Ser Pro Thr Gln Ser Pro Ala His Lys Pro Cys Ser Gly Ser Gln Lys Asp Leu 65 70 75 ao Ser Asp Val Gln Trp Tyr Met Gln Pro Arg Ser Gly Ser Pro Leu Glu Glu Ile Ser Arg Asn Ser Pro His Met Gln Ser Glu Gly Met Leu His Ile Leu Ala Pro Gln Thr Asn Ser Ile Trp Ser Tyr Ile Cys Arg Pro Arg Ile Arg Ser Pro Gln Asp Met Ala Cys Cys Ile Lys Thr Val Leu Glu Val Lys Pro Gln~Arg Asn Val Ser Cys Gly Asn Thr Ala Gln Asp Glu Gln Val Leu Leu Leu Gly Ser Thr Gly Ser Ile His Cys Pro Ser Leu Ser Cys Gln Ser Asp Val Gln Ser Pro Glu Met Thr Trp Tyr Lys Asp Gly Arg Leu Leu Pro Glu His Lys Lys Asn Pro Ile Glu Met Ala Asp Ile Tyr Val Phe Asn Gln Gly Leu Tyr Val Cys Asp Tyr Thr Gln Ser Asp Asn Val Ser Ser Trp Thr Val Arg Ala Val Val Lys Val Arg Thr Ile Gly Lys Asp Ile Asn Val Lys Pro Glu Ile Leu Asp Pro Ile Thr Asp Thr Leu Asp Val Glu Leu Gly Lys Pro Leu Thr Leu Pro Cys Arg Val Gln Phe Gly Phe Gln Arg Leu Ser Lys Pro Val Ile Lys Trp Tyr Val Lys Glu Ser Thr Gln Glu Trp Glu Met Ser Val Phe Glu Glu Lys Arg Ile Gln Ser Thr Phe Lys Asn Glu Val Ile Glu Arg Thr Ile Phe Leu Arg Glu Val Thr Gln Arg Asp Leu Ser Arg Lys Phe Val Cys 325 . 330 335 Phe Ala Gln Asn Ser Ile Gly Asn Thr Thr Arg Thr Ile Arg Leu Arg Lys Lys Glu Glu Val Val Phe Val Tyr Ile Leu Leu Gly Thr Ala Leu Met Leu Val Gly Val Leu Val Ala Ala Ala Phe Leu Tyr Trp Tyr Trp Ile Glu Val Val Leu Leu Cys Arg Thr Tyr Lys Asn Lys Asp Glu Thr Leu Gly Asp Lys Lys Glu Phe Asp Ala Phe Val Ser Tyr Ser Asn Trp Ser Ser Pro Glu Thr Asp Ala Val Gly Ser Leu Ser Glu Glu His Leu Ala Leu Asn Leu Phe Pro Glu Val Leu Glu Asp Thr Tyr Gly Tyr Arg Leu Cys Leu Leu Asp Arg Asp Val Thr Pro Gly Gly Val Tyr Ala Asp Asp Ile Val Ser Ile Ile Lys Lys Ser Arg Arg Gly Ile Phe Ile Leu Ser Pro Ser Tyr Leu Asn Gly Pro Arg Val Phe Glu Leu Gln Ala Ala Val Asn Leu Ala Leu Val Asp Gln Thr Leu Lys Leu Ile Leu Ile Lys Phe Cys Ser Phe Gln Glu Pro Glu Ser Leu Pro Tyr Leu Val Lys,~ys Ala Leu Arg Val Leu Pro Thr Val Thr Trp Lys Gly Leu Lys Ser Val His Ala Ser Ser Arg Phe Trp Thr Gln Ile Arg Tyr His Met Pro Val Lys Asn Ser Asn Arg Phe Met Phe Asn Gly Leu Arg Ile Phe Leu Lys Gly Phe Ser Pro Glu Lys Asp Leu Val Thr Gln Lys Pro Leu Glu Gly Met Pro Lys Ser Gly Asn Asp His Gly Ala Gln Asn Leu Leu Leu Tyr Ser Asp Gln Lys Arg Cys (2) INFORMATION FOR SEQ ID N0:17:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 516 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA
(ix) FEATURE:
(A) NAME/KEY: CDS
(B) LOCATION: 2..514 (ix) FEATURE:
(A) NAME/KEY: misc-feature (B) LOCATION: 374 (D) OTHER INFORMATION: /note= "nucleotides 374, 383, 396, 403, 433. 458, 459, 483, and 515 are indicated as C; each may be A, C, G, or T"
(xi) SEQUENCE DESCRIPTION: SEQ ID N0:17:

Cys Glu Leu Lys Tyr Gly Gly Phe Val Val Arg Arg Thr Thr Glu Leu Thr Val Thr Ala Pro Leu Thr Asp Lys Pro Pro Lys Leu Leu Tyr Pro Met Glu Ser Lys Leu Thr Ile Gln Glu Thr Gln Leu Gly Asp Ser AAT ACC AGA TAC GAT
GCT

Ala Leu CysArg PhePhe Gly SerGly Val Ser Asn Thr Ala Tyr Asp TTA TAC AAA TTT GAT

Pro Ile TrpMet GlyGlu Lys IleGlu Leu Asp Leu Tyr Lys Phe Asp ..

AAT GTT AGT ATT GAG

Glu Arg TrpGlu AspIle Arg ,LeuLys His Leu Asn Val Ser Ile Glu GAA GAA ATC GTG GTG

Gly Gln ValSer SerLeu Ile AspSer Glu Glu Glu Glu Ile Val Val GAC GGA TCC GAA CAA

Gly Leu AsnTyr CysTyr Val LysTrp Trp Thr Asp Gly Ser Glu Gln ACA CAG CCC GAG ATG

Pro Arg ProSer PheIle Asn SerLeu Tyr Thr Thr Gln Pro Glu Met GGA TGC GCC AAA TGG

Val Thr LeuGlu LeuGly Pro ProTrp Leu Asn Gly Cys Ala Lys Trp TCG CCA AAG GTT

Val Gly ProSer CysThr Lys Gly Ser Pro Lys Val (2) INFORMATION FOR SEQ ID N0:18:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 171 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID N0:18:
Cys Glu Leu Lys Tyr Gly Gly Phe Val Val Arg Arg Thr Thr Glu Leu Thr Val Thr Ala Pro Leu Thr Asp Lys Pro Pro Lys Leu Leu Tyr Pro Met Glu Ser Lys Leu Thr Ile Gln Glu Thr Gln Leu Gly Asp Ser Ala Asn Leu Thr Cys Arg Ala Phe Phe Gly Tyr Ser Gly Asp Val Ser Pro Leu Ile Tyr Trp Met Lys Gly Glu Lys Phe Ile Glu Asp Leu Asp Glu Asn Arg Val Trp Glu Ser Asp Ile Arg Ile Leu Lys Glu His Leu Gly Glu Gln Glu Val Ser Ile Ser Leu Ile Val Asp Ser Val Glu Glu Gly Asp Leu Gly Asn Tyr Ser Cys Tyr Val Glu Lys Trp Gln Trp Thr Pro Thr Arg Gln Pro Ser Pro Phe Ile Asn Glu Ser Leu Met Tyr Thr Val Gly Thr Cys Leu Glu Ala Leu Gly Pro Lys Pro Trp Trp Leu Asn Val Ser Gly Pro Pro Ser Lys Cys Thr Lys Val Gly (2) INFORMATION FOR SEQ ID N0:19:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 1991 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA
(ix) FEATURE:
(A) NAME/KEY: CDS
(B) LOCATION: 1..1458 (xi) SEQUENCE
DESCRIPTION:
SEQ ID N0:19:

ACG TAT GGA GTT

Glu Phe Gly Ser Cys Glu Leu Lys Gly Phe Val Arg Thr Tyr Gly Val GAA CCT CTG AAG

Arg Thr Thr Leu Thr Val Thr Ala Thr Asp Pro Pro Glu Pro Leu Lys TAT CTG ACA GAG

Lys Leu Leu Pro Met Glu Ser Lys Ile Gln Thr Gln Tyr Leu Thr Glu TCT AGA GCT GGG

Leu Gly Asp Ala Asn Leu Thr Cys Phe Phe Tyr Ser Ser Arg Ala Gly AGT ATG AAA AAA

Gly Asp Val Pro Leu Ile Tyr Trp Gly Glu Phe Ile Ser Met Lys Lys GAT GAA AGT AGA

Glu Asp Leu Glu Asn Arg Val Trp Asp Ile Ile Leu Asp Glu Ser Arg CTT TCC ATC ATT

Lys Glu His Gly Glu Gln Glu Val Ser Leu Val Asp Leu Ser Ile Ile GAA TAC TCC GTT

Ser Val Glu Gly Asp Leu Gly Asn Cys Tyr Glu Asn Glu Tyr Ser Val WO 99/19480 37 PCT/US98l20939 AAT
GGA
CGT
CGA
CAC
GCC
AGC
GTT
CTC
CTT
CAT
AAA
CGA
GAG
CTA

Gly Asn Gly Arg Arg His Ala Ser Val Leu Leu His Lys Arg Glu Leu TAC TTG CTG

Met Thr Val Glu Leu Ala Gly Gly Leu Gly Ala Ile Leu Tyr Leu Leu . ..

GTA ATC ATG

Leu Cys Leu Val Thr Ile Tyr Lys Cys Tyr Lys Ile Glu Val Ile Met TTC GAC AAT

Leu Tyr Arg Asn His Phe Gly Ala Glu Glu Leu Asp Gly Phe Asp Asn GAT GAC CAG

Lys Tyr Asp Ala Tyr Leu Ser Tyr Thr Lys Val Asp Pro Asp Asp Gln AAT ATC CTA

Trp Gln Glu Thr Gly Glu Glu Glu Arg Phe Ala Leu Glu Asn Ile Leu GAT CCA GAT

Pro Met Leu Glu Lys His Tyr Gly Tyr Lys Leu Phe Ile Asp Pro Asp GAT AGA TGT

Arg Leu Ile Pro Thr Gly Thr Tyr Ile Glu Asp Val Ala Asp Arg Cys GAT TAC GTA

Val Gln Ser Lys Arg Leu Ile Ile Val Met Thr Pro Asn Asp Tyr Val AGA CGA AAT

Val Arg Gly Trp Ser Ile Phe Glu Leu Glu Thr Arg Leu Arg Arg Asn CTT AGT GAA

Met Val Thr Gly Glu Ile Lys Val Ile Leu Ile Glu Cys Leu Ser Glu AGA CAC ACC

Leu Gly Ile Met Asn Tyr Gln Glu Val Glu Ala Leu Lys Arg His Thr AAG AAC AAG

Ile Leu Leu Thr Val Ile Lys Trp His Gly Pro Lys Cys Lys Asn Lys AAC TTT AAG

Leu Ser Lys Phe Trp Lys Arg Leu Gln Tyr Glu Met Pro Asn Phe Lys ATA GAG CAA

Arg Glu Pro Ile Thr His Glu Gln Ala Leu Asp Val Ser Ile Glu Gln CCT GCC GCG

Gly Phe Gly Glu Leu Gln Thr Val Ser Ala Ile Ser Met Pro Ala Ala WO 99/19480 3 $ PCT/US98/20939 GCC ACT CCA GAT
CTC CGT
TCT ACC

Ala Thr Ser Thr Ala Leu Ala His Asp Leu Arg Ser Ala Thr Pro Thr TCA CAA CAG CGA

Phe His Asn Thr Tyr His Met Arg Lys His Tyr Tyr Ser Gln Gln Arg , ..

CCT CCT ACC TCC

Ser Tyr Glu Tyr Asp Val Thr Gly Leu Pro Leu Thr Pro Pro Thr Ser TAC TGT CCT AAC

Ile Gly Asn Gln His Thr Asn Ile Met Thr Leu Ile Tyr Cys Pro Asn AAA TCG GAG GAG

Gly Gln Arg Pro Gln Thr Ser Arg Gln Asn Pro Asp Lys Ser Glu Glu ATC CTG TTG AGT

Ala His Thr Asn Ser Ala Pro Leu Pro Arg Glu Thr Ile Leu Leu Ser ATA TCC AGT GTG ATA TGG l4gg TGACAGAAAA GCAAGGGACA TCCCGTCCCT

Ile Ser Ser Val Ile Trp A.AAAAACATG CATTAGAATC TTTAGAACACGAGGAAAAACAGGGTCTTGT ACATATGTTT1608 GGAAAAAAAA ~1,AAAAA.AAAA 19 (2) INFORMATION FOR SEQ ID N0:20:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 486 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID N0:20:
Glu Phe Gly Thr Ser Cys Glu Leu Lys Tyr Gly Gly Phe Val Val Arg..

Arg Thr Thr Glu Leu Thr Val Thr Ala Pro Leu Thr Asp Lys Pro Pro Lys Leu Leu Tyr Pro Met Glu Ser Lys Leu Thr Ile Gln Glu Thr Gln Leu Gly Asp Ser Ala Asn Leu Thr Cys Arg Ala Phe Phe Gly Tyr Ser Gly Asp Val Ser Pro Leu Ile Tyr Trp Met Lys Gly Glu Lys Phe"tle Glu Asp Leu Asp Glu Asn Arg Val Trp Glu Ser Asp Ile Arg Ile Leu Lys Glu His Leu Gly Glu Gln Glu Val Ser Ile Sex Leu Ile Val Asp Ser Val Glu Glu Gly Asp Leu Gly Asn Tyr Ser Cys Tyr Val Glu Asn Gly Asn Gly Arg Arg His Ala Ser Val Leu Leu His Lys Arg Glu Leu Met Tyr Thr Val Glu Leu Ala Gly Gly Leu Gly Ala Ile Leu Leu Leu Leu Val Cys Leu Val Thr Ile Tyr Lys Cys Tyr Lys Ile Glu Ile Met Leu Phe Tyr Arg Asn His Phe Gly Ala Glu Glu Leu Asp Gly Asp Asn Lys Asp Tyr Asp Ala Tyr Leu Ser Tyr Thr Lys Val Asp Pro Asp Gln Trp Asn Gln Glu Thr Gly Glu Glu Glu Arg Phe Ala Leu Glu Ile Leu Pro Asp Met Leu Glu Lys His Tyr Gly Tyr Lys Leu Phe Ile Pro Asp Arg Asp Leu Ile Pro Thr Gly Thr Tyr Ile Glu Asp Val Ala Arg Cys Val Asp Gln Ser Lys Arg Leu Ile Ile Val Met Thr Pro Asn Tyr Val Val Arg Arg Gly Trp Ser Ile Phe Glu Leu Glu Thr Arg Leu Arg Asn Met Leu Val Thr Gly Glu Ile Lys Val Ile Leu Ile Glu Cys Ser Glu Leu Arg Gly Ile Met Asn Tyr Gln Glu Val Glu Ala Leu Lys His Thr Ile Lys Leu Leu Thr Val Ile Lys Trp His Gly Pro Lys Cys Asn Lys Leu Asn Ser Lys Phe Trp Lys Arg Leu Gln Tyr Glu Met Pro Phe Lys Arg Ile Glu Pro Ile Thr His Glu Gln Ala Leu Asp Val Ser Glu Gln Gly Pro Phe Gly Glu Leu Gln Thr Val Ser Ala Ile Ser Met Ala Ala Ala Thr Ser Thr Ala Leu Ala Thr Ala His Pro Asp Leu Arg Ser Thr Phe His Asn Thr Tyr His Ser Gln Met Arg Gln Lys His Tyr T~rr.Arg Ser Tyr Glu Tyr Asp Val Pro Pro Thr Gly Thr Leu Pro Leu Thr Ser Ile Gly Asn Gln His Thr Tyr Cys Asn Ile Pro Met Thr Leu Ile Asn Gly Gln Arg Pro Gln Thr Lys Ser Ser Arg Glu Gln Asn Pro Asp Glu Ala His Thr Asn Ser Ala Ile Leu Pro Leu Leu Pro Arg Glu Thr Ser Ile Ser Ser Val Ile Trp (2) INFORMATION FOR SEQ ID N0:21:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 570 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: not relevant (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID N0:21:
Met Gly Leu Leu Trp Tyr Leu Met Ser Leu Ser Phe Tyr Gly Ile Leu Gln Ser His Ala Ser Glu Arg Cys Asp Asp Trp Gly Leu Asp Thr Met Arg Gln Ile Gln Val Phe Glu Asp Glu Pro Ala Arg Ile Lys Cys Pro Leu Phe Glu His Phe Leu Lys Tyr Asn Tyr Ser Thr Ala His Ser Ser Gly Leu Thr Leu Ile Trp Tyr Trp Thr Arg Gln Asp Arg Asp Leu Glu Glu Pro Ile Asn Phe Arg Leu Pro Glu Asn Arg Ile Ser Lys Glu Lys Asp Val Leu Trp Phe Arg Pro Thr Leu Leu Asn Asp Thr Gly Asn Tyr Thr Cys Met Leu Arg Asn Thr Thr Tyr Cys Ser Lys Val Ala Phe Pro Leu Glu Val Val Gln Lys Asp Ser Cys Phe Asn Ser Ala Met Arg Phe Pro Val His Lys Met Tyr Ile Glu His Gly Ile His Lys Ile Thr Cys Pro Asn Val Asp Gly Tyr Phe Pro Ser Ser Val Lys Pro Sex Val Thr Trp Tyr Lys Gly Cys Thr Glu Ile Val Asp Phe His Asn Val Leu Pro Glu Gly Met Asn Leu Ser Phe Phe Ile Pro Leu Val Ser Asn Asn Gly Asn Tyr Thr Cys Val Val Thr Tyr Pro Glu Asn Gly Arg Leu Phe His Leu Thr Arg Thr Val Thr Val Lys Val Val Gly Ser Pro Lys Asp Ala Leu Pro Pro Gln Ile Tyr Ser Pro Asn Asp Arg Val Val Tyr GIu Lys Glu Pro Gly Glu Glu Leu Val Ile Pro Cys Lys Val Tyr Phe Ser Phe Ile Met Asp Ser His Asn Glu Val Trp Trp Thr Ile Asp Gly Lys Lys Pro Asp Asp Val Thr Val Asp Ile Thr Ile Asn Glu Ser Val Ser Tyr Ser Ser Thr Glu Asp Glu Thr Arg Thr Gln Ile Leu Ser Ile Lys Lys Val Thr Pro Glu Asp Leu Arg Arg Asn Tyr Val Cys His Ala Arg Asn Thr Lys Gly Glu Ala Glu Gln Ala Ala Lys Val Lys Gln Lys Val Ile Pro Pro Arg Tyr Thr Val Glu Leu Ala Cys Gly Phe Gly Ala Thr Val Phe Leu Val Val Val Leu Ile Val Val Tyr His Val Tyr Trp Leu Glu Met Val Leu Phe Tyr Arg Ala His Phe Gly Thr Asp Glu Thr Ile Leu Asp Gly Lys Glu Tyr Asp Ile Tyr Val Ser Tyr Ala Arg Asn Val Glu Glu Glu Glu Phe Val Leu Leu Thr Leu Arg Gly Val Leu Glu Asn Glu Phe Gly Tyr Lys Leu Cys Ile Phe Asp Arg Asp Ser Leu Pro Gly Gly Ile Val Thr Asp Glu Thr Leu Ser Phe Ile Gln Lys Ser Arg Arg Leu Leu Val Val Leu Ser Pro Asn Tyr Val Leu Gln Gly Thr Gln Ala Leu Leu Glu Leu Lys Ala Gly Leu Glu Asn Met Ala Ser Arg Gly Asn Ile Asn Val Ile Leu Val Gln Tyr Lys Ala Val Lys Asp Met ~y$,Val Lys Glu Leu Lys Arg Ala Lys Thr Val Leu Thr Val Ile Lys Trp Lys Gly Glu Lys Ser Lys Tyr Pro Gln Gly Arg Phe Trp Lys Gln Leu Gln Val Ala Met Pro Val Lys Lys Ser Pro Arg Trp Ser Ser Asn Asp Lys Gln Gly Leu Ser Tyr Sex Ser Leu Lys Asn Val (2) INFORMATION FOR SEQ ID N0:22:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 562 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: not relevant (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID N0:22:
Met Trp Ser Leu Leu Leu Cys Gly Leu Ser Ile Ala Leu Pro Leu Ser Val Thr Ala Asp Gly Cys Lys Asp Ile Phe Met Lys Asn Glu Ile Leu Ser Ala Ser Gln Pro Phe Ala Phe Asn Cys Thr Phe Pro Pro Ile Thr Ser Gly Glu Val Ser Val Thr Trp Tyr Lys Asn Ser Ser Lys Ile Pro Val Ser Lys Ile Ile Gln Ser Arg Ile His Gln Asp Glu Thr Trp Ile 65 70 75 g0 Leu Phe Leu Pro Met Glu Trp Gly Asp Ser Gly Val Tyr Gln Cys Val Ile Lys Gly Arg Asp Ser Cys His Arg Ile His Val Asn Leu Thr Val Phe Glu Lys His Trp Cys Asp Thr Ser Ile Gly Gly Leu Pro Asn Leu Ser Asp Glu Tyr Lys Gln Ile Leu His Leu Gly Lys Asp Asp Ser Leu Thr Cys His Leu His Phe Pro Lys Ser Cys Val Leu Gly Pro Ile Lys Trp Tyr Lys Asp Cys Asn Glu Ile Lys Gly Glu Arg Phe Thr Val Leu Glu Thr Arg Leu Leu Val Ser Asn Val Ser Ala Glu Asp Arq, Gly Asn Tyr Ala Cys Gln Ala Ile Leu Thr His Ser Gly Lys Gln Tyr Glu Val Leu Asn Gly Ile Thr Val Ser Ile Thr Glu Arg Ala Gly Tyr Gly Gly Ser Val Pro Lys Ile Ile Tyr Pro Lys Asn His Ser Ile Glu Val Gln Leu Gly Thr Thr Leu Ile Val Asp Cys Asn Val Thr Asp Thr Lys Asp Asn Thr Asn Leu Arg Cys Trp Arg Val Asn Asn Thr Leu Val Asp Asp Tyr Tyr Asp Glu Ser Lys Arg Ile Arg Glu Gly Val Glu Thr His Val Ser Phe Arg Glu His Asn Leu Tyr Thr Val Asn Ile Thr Phe Leu Glu Val Lys Met Glu Asp Tyr Gly Leu Pro Phe Met Cys His Ala Gly Val Ser Thr Ala Tyr Ile Ile Leu Gln Leu Pro Ala Pro Asp Phe Arg Ala Tyr Leu Ile Gly Gly Leu Ile Ala Leu Val Ala Val Ala Val Ser Val Val Tyr Ile Tyr Asn Ile Phe Lys Ile Asp Ile Val Leu Trp Tyr Arg Ser Ala Phe His Ser Thr Glu Thr Ile~Val Asp Gly Lys Leu Tyr Asp Ala Tyr Val Leu Tyr Pro Lys Pro His Lys Glu Ser Gln Arg His Ala Val Asp Ala Leu Val Leu Asn Ile Leu Pro Glu Val Leu Glu Arg Gln Cys Gly Tyr Lys Leu Phe Ile Phe Gly Arg Asp Glu Phe Pro Gly Gln Ala Val Ala Asn Val Ile Asp Glu Asn Val Lys Leu Cys Arg Arg Leu Ile Val Ile Val Val Pro Glu Ser Leu Gly Phe Gly Leu Leu Lys Asn Leu Ser Glu Glu Gln Ile Ala Val Tyr Ser Ala Leu Ile Gln Asp Gly Met Lys Val Ile Leu Ile Glu Leu Glu Lys Ile Glu Asp Tyr Thr Val 485 ~ 490 495 Met Pro Glu Ser Ile Gln Tyr Ile Lys Gln Lys His Gly Ala Ile Arg Trp His Gly Asp Phe Thr Glu Gln Ser Gln Cys Met Lys Thr Lys Phe Trp Lys Thr Val Arg Tyr His Met Pro Pro Arg Arg Cys Arg Pro Phe Leu Arg Ser Thr Cys Arg Ser Thr His Leu Cys Thr Ala Pro Gln Ala Gln Asn (2) INFORMATION FOR SEQ ID N0:23:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 561 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: not relevant (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID N0:23:
Met Gly Met Pro Pro Leu Leu Phe Cys Trp Val Ser Phe Val Leu Pro Leu Phe Val Ala Ala Gly Asn Cys Thr Asp Val Tyr Met His His Glu Met Ile Ser Glu Gly Gln Pro Phe Pro Phe Asn Cys Thr Tyr Pro Pro Val Thr Asn Gly Ala Val Asn Leu Thr Trp His Arg Thr Pro Ser Lys Ser Pro Ile Ser Ile Asn Arg His Val Arg Ile His Gln Asp Gln Ser 65 70 75 gp Trp Ile Leu Phe Leu Pro Leu Ala Leu Glu Asp Ser Gly Ile Tyr Gln Cys Val Ile Lys Asp Ala His Ser Cys Tyr Arg Ile Ala Ile Asn Leu Thr Val Phe Arg Lys His Trp Cys Asp Ser Ser Asn Glu Glu Ser Ser Ile Asn Ser Ser Asp Glu Tyr Gln Gln Trp Leu Pro Ile Gly Lys Ser Gly Ser Leu Thr Cys His Leu Tyr Phe Pro Glu Ser Cys Val Leu Asp Ser Ile Lys Trp Tyr Lys Gly Cys Glu Glu Ile Lys Val Ser Lys Lys Phe Cys Pro Thr Gly Thr Lys Leu Leu Val Asn Asn Ile Asp Val Glu Asp Ser Gly Ser Tyr Ala Cys Ser Ala Arg Leu Thr His Leu.Gly Arg Ile Phe Thr Val Arg Asn Tyr Ile Ala Val Asn Thr Lys Glu Val Gly Ser Gly Gly Arg Ile Pro Asn Ile Thr Tyr Pro Lys Asn Asn Ser Ile Glu Val Gln Leu Gly Ser Thr Leu Ile Val Asp Cys Asn Ile Thr Asp Thr Lys Glu Asn Thr Asn Leu Arg Cys Trp Arg Val Asn Asn Thr Leu Val Asp Asp Tyr Tyr Asn Asp Phe Lys Arg Ile Gln Glu Gly Ile Glu Thr Asn Leu Ser Leu Arg Asn His Ile Leu Tyr Thr Val Asn Ile Thr Phe Leu Glu Val Lys Met Glu Asp Tyr Gly His Pro Phe Thr Cys His Ala Ala Val Ser Ala Ala Tyr Ile Ile Leu Lys Arg Pro Ala Pro Asp Phe Arg Ala Tyr Leu Ile Gly Gly Leu Met Ala Phe Leu Leu Leu Ala Val Ser Ile Leu Tyr Ile Tyr Asn Thr Phe Lys Val Asp Ile Val Leu Trp Tyr Arg Ser Thr Phe His Thr Ala Gln Ala Pro Asp Asp Glu Lys Leu Tyr Asp Ala Tyr Val Leu Tyr Pro Lys Tyr Pro Arg Glu Ser Gln Gly His Asp Val Asp Thr Leu Val Leu Lys Ile Leu Pro Glu Val Leu Glu Lys Gln Cys Gly Tyr Lys Leu Phe Ile Phe Gly Arg Asp Glu Phe Pro Gly Gln Ala Val Ala Ser Val Ile Asp Glu Asn Ile Lys Leu Cys Arg Arg Leu Met Val Leu Val Ala Pro Glu Thr Ser Ser Phe Ser Phe Leu Lys Asn Leu Thr Glu Glu Gln Ile Ala Val Tyr Asn Ala Leu Val 465 470 475 .. 480 Gln Asp Gly Met Lys Val Ile Leu Ile Glu Leu Glu Arg Val Lys Asp Tyr Thr Pro SerIleGln TyrIle Arg Lys His Ser Met Glu Gln Gly Ala Ile GlnTrp GlyAspPhe ThrGlu Gln Gln Cys Lys Asp Ala Ala Thr Lys PheTrp LysValArg TyrHis Met Pro Ar Arg Tyr Lys Pro Pro Ala SerPro ValGlnLeu LeuGly His Pro Arg Pro Pro Thr Ile Gly (2) INFORMATION FOR SEQ ID N0:24:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 567 amino acids (B) TYPE: amino acid - (C) STRANDEDNESS: not relevant (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID N0:24:
Met Ile Asp Arg Gln Arg Met Gly Leu Trp Ala Leu Ala Ile Leu Thr Leu Pro Met Tyr Leu Thr Val Thr Glu Gly Ser Lys Ser Ser Trp Gly Leu Glu Asn Glu Ala Leu Ile Val Arg Cys Pro Gln Arg Gly Arg Ser Thr Tyr Pro Val Glu Trp Tyr Tyr Ser Asp Thr Asn Glu Ser Ile Pro Thr Gln Lys Arg Asn Arg Ile Phe Val Ser Arg Asp Arg Leu Lys Phe 65 70 75 , 80 Leu Pro Ala Arg Val Glu Asp Ser Gly Ile Tyr Ala Cys Val Ile Arg Ser Pro Asn Leu Asn Lys Thr Gly Tyr Leu Asn Val Thr Ile His Lys Lys Pro Pro Ser Cys Asn Ile Pro Asp Tyr Leu Met Tyr Ser Thr Val Arg Gly Ser Asp Lys Asn Phe Lys Ile Thr Cys Pro Thr Ile Asp Leu Tyr Asn Trp Thr Ala Pro Val Gln Trp Phe Lys Asn Cys Lys Ala Leu 145 250 155 °-160 Gln Glu Pro Arg Phe Arg Ala His Arg Ser Tyr Leu Phe Ile Asp Asn Val Thr His Asp Asp Glu Gly Asp Tyr Thr Cys Gln Phe Thr His Ala Glu Asn Gly Thr Asn Tyr Ile Val Thr Ala Thr Arg Ser Phe Thr Val Glu Glu Lys Gly Phe Ser Met Phe Pro Val Ile Thr Asn ,Prq Pro Tyr Asn His Thr Met Glu Val Glu Ile Gly Lys Pro Ala Ser Ile Ala Cys Ser Ala Cys Phe Gly Lys Gly Ser His Phe Leu Ala Asp Val Leu Trp Gln Ile Asn Lys Thr Val Val Gly Asn Phe Gly Glu Ala Arg ,Ile Gln Glu Glu Glu Gly Arg Asn Glu Ser Ser Ser Asn Asp Met Asp Cys Leu Thr Ser Val Leu Arg Ile Thr Gly Val Thr Glu Lys Asp Leu Ser Leu Glu Tyr Asp Cys Leu Ala Leu Asn Leu His Gly Met Ile Arg His Thr Ile Arg Leu Arg Arg Lys Gln Pro Ile Asp His Arg Ser Ile Tyr Tyr Ile Val Ala Gly Cys Ser Leu Leu Leu Met Phe Ile Asn Val Leu Val Ile Val Leu Lys Val Phe Trp Ile Glu Val Ala Leu Phe Trp Arg Asp Ile Val Thr Pro Tyr Lys Thr Arg Asn Asp Gly Lys Leu Tyr Asp Ala Tyr Ile Ile Tyr Pro Arg Val Phe Arg Gly Ser Ala Ala Gly Thr His Ser Val Glu Tyr Phe Val His His Thr Leu Pro Asp Val Leu Glu Asn Lys Cys Gly Tyr Lys Leu Cys Ile Tyr Gly Arg Asp Leu Leu Pro Gly Gln Asp Ala Ala Thr Val Val Glu Ser Ser Ile Gln Asn Ser Arg Arg Gln Val Phe Val Leu Ala Pro His Met Met His Ser Lys Glu Phe Ala Tyr Glu Gln Glu Ile Ala Leu His Ser Ala Leu Ile Gln Asn Asn Ser Lys Val Ile Leu Ile Glu Met Glu Pro Leu Gly Glu Ala Ser Arg Leu Gln Val Gly Asp Leu Gln Asp Ser Leu Gln His Leu Val Lys Ile Gln Gly Thr Ile Lys Trp Arg Glu Asp His Val Ala Asp Lys Gln Ser Leu Ser Ser Lys Phe Trp Lys His Val Arg Tyr Gln Met Pro Val Pro Glu Arg Ala Ser Lys Thr Ala Ser Val Ala Ala Pro Leu Ser (',,lX,Lys Ala Cys Leu Asp Leu Lys His Phe (2) INFORMATION FOR SEQ ID N0:25:
(i) SEQUENCE CHARACTERISTICS: , (A) LENGTH: 328 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: not relevant (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID N0:25:
Met Gly Phe Trp Ile Leu Ala Ile Leu Thr Ile Leu Met Tyr Ser Thr Ala Ala Lys Phe Ser Lys Gln Ser Trp Gly Leu Glu Asn Glu Ala Leu Ile Val Arg Cys Pro Arg Gln Gly Lys Pro Ser Tyr Thr Val Asp Trp Tyr Tyr Ser Gln Thr Asn Lys Ser Ile Pro Thr Gln Glu Arg Asn Arg Val Phe Ala Ser Gly Gln Leu Leu Lys Phe Leu Pro Ala Glu Val Ala 65 70 75 g0 Asp Ser Gly Ile Tyr Thr Cys Ile Val Arg Ser Pro Thr Phe Asn Arg Thr Gly Tyr Ala Asn Val Thr Ile Tyr Lys Lys Gln Ser Asp Cys Asn Val Pro Asp Tyr Leu Met Tyr Ser Thr Val Ser Gly Ser Glu Lys Asn Ser Lys Ile Tyr Cys Pro Thr Ile Asp Leu Tyr Asn Trp Thr Ala Pro Leu Glu Trp Phe Lys Asn Cys Gln Ala Leu Gln Gly Ser Arg Tyr Arg Ala His Lys Ser Phe Leu Val Ile Asp Asn Val Met Thr Glu Asp Ala Gly Asp Tyr Thr Cys Lys Phe Ile His Asn Glu Asn Gly Ala Asn Tyr Ser Val Thr Ala Thr Arg Ser Phe Thr Val Lys Asp Glu Gln Gly Phe Ser Leu Phe Pro Val Ile Gly Ala Pro Ala Gln Asn Glu Ile Lys Glu Val Glu Ile Gly Lys Asn Ala Asn Leu Thr Cys Ser Ala ~y~ Phe GIy Lys Gly Thr Gln Phe Leu Ala Ala Val Leu Trp Gln Leu Asn Gly Thr Lys Ile Thr Asp Phe Gly Glu Pro Arg Ile Gln Gln Glu Glu Gly Gln Asn Gln Ser Phe Ser Asn Gly Leu Ala Cys Leu Asp Met Val Leu Arg Ile Ala Asp Val Lys Glu Glu Asp Leu Leu Leu~Gln Tyr Asp Cys Leu Ala Leu Asn Leu His Gly Leu Arg Arg His Thr Val Arg Leu Ser Arg Lys Asn Pro Ser Lys Glu Cys Phe (2) INFORMATION FOR SEQ ID N0:26:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 398 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: not relevant (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID N0:26:
Met Leu Arg Leu Tyr Val Leu Val Met Gly Val Ser Ala Phe Thr Leu Gln Pro Ala Ala His Thr Gly Ala Ala Arg Ser Cys Arg Phe Arg Gly Arg His Tyr Lys Arg Glu Phe Arg Leu Glu Gly Glu Pro Val Ala Leu Arg Cys Pro Gln Val Pro Tyr Trp Leu Trp Ala Ser Val Ser Pro Arg Ile Asn Leu Thr Trp His Lys Asn Asp Ser Ala Arg Thr Val Pro Gly Glu Glu Glu Thr Arg Met Trp Ala Gln Asp Gly Ala Leu Trp Leu Leu 85 9p 95..
Pro Ala Leu Gln Glu Asp Ser Gly Thr Tyr Val Cys Thr Thr Arg Asn Ala Ser Tyr Cys Asp Lys Met Ser Ile Glu Leu Arg Val Phe Glu Asn Thr Asp Ala Phe Leu Pro Phe Ile Ser Tyr Pro Gln Ile Leu Thr Leu Ser Thr Ser Gly Val Leu Val Cys Pro Asp Leu Ser Glu ~h~ Thr Arg Asp Lys Thr Asp Val Lys Ile Gln Trp Tyr Lys Asp Ser Leu Leu Leu Asp Lys Asp Asn Glu Lys Phe Leu Ser Val Arg Gly Thr Thr His Leu Leu Val His Asp Val Ala Leu Glu Asp Ala Gly Tyr Tyr Arg Cys Val Leu Thr Phe Ala His Glu Gly Gln Gln Tyr Asn-Ile Thr Arg Ser Ile Glu Leu Arg Ile Lys Lys Lys Lys Glu Glu Thr Ile Pro Val Ile Ile Ser Pro Leu Lys Thr Ile Ser Ala Ser Leu Gly Ser Arg Leu Thr Ile Pro Cys Lys Val Phe Leu Gly Thr Gly Thr Pro Leu Thr Thr Met Leu Trp Trp Thr Ala Asn Asp Thr His Ile Glu Ser Ala Tyr Pro Gly Gly Arg Val Thr Glu Gly Pro Arg Gln Glu Tyr Ser Glu Asn Asn Glu Asn Tyr Ile Glu Val Pro Leu Ile Phe Asp Pro Val Thr Arg Glu Asp Leu His Met Asp Phe Lys Cys Val Val His Asn Thr Leu Ser Phe Gln Thr Leu Arg Thr Thr Val Lys Glu Ala Ser Ser Thr Phe Ser Trp Gly Ile Val Leu Ala Pro Leu Ser Leu Ala Phe Leu Val Leu Gly Gly Ile Trp Met His Arg Arg Cys Lys His Arg Thr Gly Lys Ala Asp Gly Leu Thr Val Leu Trp Pro His His Gln Asp Phe Gln Ser Tyr Pro Lys (2) INFORMATION FOR SEQ ID N0:27:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 410 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: not relevant (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID N0:27:
Met Phe Ile Leu Leu Val Leu Val Thr Gly Val Ser Ala Phe Thr Thr 1 5 10 ~, " 15 Pro Thr Val Val His Thr Gly Lys Val Ser Glu Ser Pro Ile Thr Ser Glu Lys Pro Thr Val His Gly Asp Asn Cys Gln Phe Arg Gly Arg Glu Phe Lys Ser Glu Leu Arg Leu Glu Gly Glu Pro Val Val Leu Arg Cys Pro Leu Ala Pro His Ser Asp Ile Ser Ser Ser Ser His Ser Phe Leu Thr Trp Ser Lys Leu Asp Ser Ser Gln Leu Ile Pro Arg Asp Glu Pro Arg Met Trp Val Lys Gly Asn Ile Leu Trp Ile Leu Pro Ala Val Gln Gln Asp Ser Gly Thr Tyr Ile Cys Thr Phe Arg Asn Ala Ser His Cys Glu Gln Met Ser Val Glu Leu Lys Val Phe Lys Asn Thr Glu Ala Ser Leu Pro His Val Ser Tyr Leu Gln Ile Ser Ala Leu Ser Thr Thr Gly Leu Leu Val Cys Pro Asp Leu Lys Glu Phe Ile Ser Ser Asn Ala Asp Gly Lys Ile Gln Trp Tyr Lys Gly Ala Ile Leu Leu Asp Lys Gly Asn Lys Glu Phe Leu Ser Ala Gly Asp Pro Thr Arg Leu Leu Ile Ser Asn Thr Ser Met Asp Asp Ala Gly Tyr Tyr Arg Cys Val Met Thr Phe Thr Tyr Asn Gly Gln Glu Tyr Asn Ile Thr Arg Asn Ile Glu Leu Arg Val Lys Gly Thr Thr Thr Glu Pro Ile Pro Val Ile Ile Ser Pro Leu Glu Thr Ile Pro Ala Ser Leu Gly Ser Arg Leu Ile Val Pro Cys Lys Val Phe Leu Gly Thr Gly Thr Ser Ser Asn Thr Ile Val Trp Trp Leu Ala Asn Ser Thr Phe Ile Ser Ala Ala Tyr Pro Arg Gly Arg Val Thr Glu Gly Leu His His Gln Tyr Ser Glu Asn Asp Glu Asn Tyr Val Glu Val Ser Leu Ile Phe Asp Pro Val Thr Arg Glu Asp Leu His Thr Asp Phe Lys Cys Val Ala Ser Asn Pro Arg Ser Ser Gln Ser Leu His Thr Thr Val Lys Glu Val Ser Ser Thr Phe Ser Trp Ser Ile Ala Leu Ala Pro Leu Ser Leu Ile Ile Leu Val Val Gly Ala Ile Trp Met Arg Arg Arg Cys Lys Arg Arg Ala Gly Lys Thr Tyr Gly Leu Thr Lys Leu Arg Thr 385 390 395 . 400 Asp Asn Gln Asp Phe Pro Ser Ser Pro Asn (2) INFORMATION FOR SEQ ID N0:28:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 541 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: not relevant (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID N0:28:
Met Asn Cys Arg Glu Leu Pro Leu Thr Leu Trp Val Leu Ile Ser Val Ser Thr Ala Glu Ser Cys Thr Ser Arg Pro His Ile Thr Val Val Glu Gly Glu Pro Phe Tyr Leu Lys His Cys Ser Cys Ser Leu Ala His Glu Ile Glu Thr Thr Thr Lys Ser Trp Tyr Lys Ser Ser Gly Ser Gln Glu His Val Glu Leu Asn Pro Arg Ser Ser Ser Arg Ile Ala Leu His Asp Cys Val Leu Glu Phe Trp Pro Val Glu Leu Asn Asp Thr Gly Ser Tyr Phe Phe Gln Met Lys Asn Tyr Thr Gln Lys Trp Lys Leu Asn Val Ile loo l05 llo Arg Arg Asn Lys His Ser Cys Phe Thr Glu Arg Gln Val Thr Ser Lys Ile Val Glu Val Lys Lys Phe Phe Gln Ile Thr Cys Glu Asn Ser~Tyr Tyr Gln Thr Leu Val Asn Ser Thr Ser Leu Tyr Lys Asn Cys Lys Lys Leu Leu Leu Glu Asn Asn Lys Asn Pro Thr Ile Lys Lys Asn Ala Glu Phe Glu Asp Gln Gly Tyr Tyr Ser Cys Val His Phe Leu His His Asn Gly Lys Leu Phe Asn Ile Thr Lys Thr Phe Asn Ile Thr Zle,Va1 Glu Asp Arg Ser Asn Ile Val Pro Val Leu Leu Gly Pro Lys Leu Asn His Val Ala Val Glu Leu Gly Lys Asn Val Arg Leu Asn Cys Ser Ala Leu Leu Asn Glu Glu Asp Val Ile Tyr Trp Met Phe Gly Glu Glu Asn Gly Ser Asp Pro Asn Ile His Glu Glu Lys Glu Met Arg Ile Met Thr Pro Glu Gly Lys Trp His Ala Ser Lys Val Leu Arg Ile Glu Asn Ile Gly Glu Ser Asn Leu Asn Val Leu Tyr Asn Cys Thr Val Ala Ser Thr Gly Gly Thr Asp Thr Lys Ser Phe Ile Leu Val Arg Lys Ala Asp Met Ala Asp Ile Pro Gly His Val Phe Thr Arg Gly Met Ile Ile Ala Val Leu Ile Leu Val Ala Val Val Cys Leu Val Thr Val Cys Val Ile Tyr Arg Val Asp Leu Val Leu Phe Tyr Arg His Leu Thr Arg Arg Asp Glu Thr Leu Thr Asp Gly Lys Thr Tyr Asp Ala Phe Val Ser Tyr Leu Lys Glu Cys Arg Pro Glu Asn Gly Glu Glu His Thr Phe Ala Val Glu Ile Leu Pro Arg Val Leu Glu Lys His Phe Gly Tyr Lys Leu Cys Ile Phe Glu Arg Asp Val Val Pro Gly Gly Ala Val Val Asp Glu Ile His Ser Leu Ile Glu Lys Ser Arg Arg Leu Ile Ile Val Leu Ser Lys Ser Tyr Met Ser Asn Glu Val Arg Tyr Glu Leu Glu Ser Gly Leu His Glu Ala Leu Val Glu Arg Lys Ile Lys Ile Ile Leu Ile Glu Phe Thr Pro Val Thr 465 470 475 ~~ -48o-Asp Phe Thr Phe Leu Pro Gln Ser Leu Lys Leu Leu Lys Ser His Arg WO 99/194$0 PCT/US98/20939 Val Leu Lys Trp Lys Ala Asp Lys Ser Leu Ser Tyr Asn Ser Arg Phe Trp Lys Asn Leu Leu Tyr Leu Met Pro Ala Lys Thr Val Lys Pro Gly Arg Asp Glu Pro Glu Val Leu Pro Val Leu Ser Glu Ser " "

(2) INFORMATION FOR SEQ ID N0:29:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 537 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: not relevant , (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID N0:29:
Met His His Glu Glu Leu Ile Leu Thr Leu Cys Ile Leu Ile Val Lys 1 ° 5 10 15 Ser Ala Ser Lys Ser Cys Ile His Arg Ser Gln I).e His Val Val Glu Gly Glu Pro Phe Tyr Leu Lys Pro Cys Gly Ile Ser Ala Pro Val His Arg Asn Glu Thr Ala Thr Met Arg Trp Phe Lys Gly Ser Ala Ser His Glu Tyr Arg Glu Leu Asn Asn Arg Ser Ser Pro Arg Val Thr Phe His Asp His Thr Leu Glu Phe Trp Pro Val Glu Met Glu Asp Glu Gly Thr Tyr Ile Ser Gln Val Gly Asn Asp Arg Arg Asn Trp Thr Leu Asn Val Thr Lys Arg Asn Lys His Ser Cys Phe Ser Asp Lys Leu Val Thr Ser Arg Asp Val Glu Val Asn Lys Ser Leu His Ile Thr Cys Lys Asn Pro Asn Tyr Glu Glu Leu Ile Gln Asp Thr Trp Leu Tyr Lys Asn Cys Lys Glu Ile Ser Lys Thr Pro Arg Ile Leu Lys Asp Ala Glu Phe Gly Asp Glu Gly Tyr Tyr Ser Cys Val Phe Ser Val His His Asn Gly Thr Arg Tyr Asn Ile Thr Lys Thr Val Asn Ile Thr Val Ile Glu Gly Arg Ser Lys Val Thr Pro Ala Ile Leu Gly Pro Lys Cys Glu Lys Val Gly Val Glu Leu Gly Lys Asp Val Glu Leu Asn Cys Ser Ala Ser Leu Asn Lys Asp Asp Leu Phe Tyr Trp Ser Ile Arg Lys Glu Asp Ser Se ~ Asp Pro Asn Val Gln Glu Asp Arg Lys Glu Thr Thr Thr Trp Ile Ser Glu Gly Lys Leu His Ala Ser Lys Ile Leu Arg Phe Gln Lys Ile Thr Glu Asn Tyr Leu Asn Val Leu Tyr Asn Cys Thr Val Ala Asn Glu Glu Ala Iie Asp Thr Lys Ser Phe Val Leu Val Arg Lys Glu Ile Pro Asp Ile Pro Gly His Val Phe Thr Gly Gly Val Thr Val Leu Val Leu Ala Ser Val Ala Ala Val Cys Ile Val Ile Leu Cys Val Ile Tyr Lys Val Asp Leu Val Leu Phe Tyr Arg Arg Ile Ala Glu Arg Asp Glu Thr Leu Thr Asp Gly Lys Thr Tyr Asp Ala Phe Val Ser Tyr Leu Lys Glu Cys His Pro Glu Asn Lys Glu Glu Tyr Thr Phe Ala Val Glu Thr Leu Pro Arg Val Leu Glu Lys Gln Phe Gly Tyr Lys Leu Cys Ile Phe Glu Arg Asp Val Val Pro Gly Gly Ala Val Val Glu Glu Ile His Ser Leu Ile Glu Lys Ser Arg erg Leu Ile Ile Val Leu Ser Gln Ser Tyr Leu Thr Asn Gly Ala Arg Arg Glu Leu Glu Ser Gly Leu His Glu Ala Leu Val Glu Arg Lys Ile Lys Ile Ile Leu Ile Glu Phe Thr Pro Ala Ser Asn Ile Thr 465 470 475 . 480 Phe Leu Pro Pro Ser Leu Lys Leu Leu Lys Ser Tyr Arg Val Leu Lys Trp Arg Ala Asp Ser Pro Ser Met Asn Ser Arg Phe Trp Lys Asn Leu Val Tyr Leu Met Pro Ala Lys Ala Val Lys Pro Trp Arg Glu Glu Ser Glu Ala Arg Ser Val Leu Ser Ala Pro (2) INFORMATION FOR SEQ ID N0:30:

(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 576 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: not relevant (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID N0:30:
Met Glu Asn Met Lys Val Leu Leu Gly Leu Ile Cys Leu Met jlal Pro Leu Leu Ser Leu Glu Ile Asp Val Cys Thr Glu Tyr Pro Asn Gln Ile Val Leu Phe Leu Ser Val Asn Glu Ile Asp Ile Arg Lys Cys Pro Leu Thr Pro Asn Lys Met His Gly Asp Thr Ile Ile Trp Tyr Lys Asn Asp Ser Lys Thr Pro Ile Ser Ala Asp Arg Asp Ser Arg Ile His Gln Gln Asn Glu His Leu Trp Phe Val Pro Ala Lys Val Glu Asp Ser Gly Tyr Tyr Tyr Cys Ile Val Arg Asn Ser Thr Tyr Cys Leu Lys Thr Lys Val Thr Val Thr Val Leu Glu Asn Asp Pro Gly Leu Cys Tyr Ser Thr Gln Ala Thr Phe Pro Gln Arg Leu His Ile Ala Gly Asp Gly Ser Leu Val Cys Pro Tyr Val Ser Tyr Phe Lys Asp Glu Asn Asn Glu Leu Pro Glu Val Gln Trp Tyr Lys Asn Cys Lys Pro Leu Leu Leu Asp Asn Val Ser Phe Phe Gly Val Lys Asp Lys Leu Leu Val Arg Asn Val Ala Glu Glu His Arg Gly Asp Tyr Ile Cys Arg Met Ser Tyr Thr Phe Arg Gly Lys Gln Tyr Pro Val Thr Arg Val Ile Gln Phe Ile Thr Ile Asp Glu Asn Lys Arg Asp Arg Pro Val Ile Leu Ser Pro Arg Asn Glu Thr Ile Glu Ala Asp Pro Gly Ser Met Ile Gln Leu Ile Cys Asn Val Thr Gly Gln Phe Ser Asp Leu Val Tyr Trp Lys Trp Asn Gly Ser Glu Ile Glu Trp Asn Asp Pro Phe Leu Ala Glu Asp Tyr Gln Phe Val Glu His Pro Ser Thr Lys Arg Lys Tyr Thr Leu Ile Thr Thr Leu Asn Ile Ser Glu Val Lys Ser Gln Phe Tyr Arg Tyr Pro Phe Ile Cys Val Val Lys Asn Thr Asn Ile Phe Glu Ser Ala His Val Gln Leu Ile Tyr Pro Val Pro Asp Phe Lys Asn Tyr Leu Ile Gly Gly Phe Ile Ile Leu Thr Ala Thr Ile 340 345 350 .
Val Cys Cys Val Cys Ile Tyr Lys Val Phe Lys Val Asp Ile Val Leu Trp Tyr Arg Asp Ser Cys Ser Gly Phe Leu Pro Ser Lys Ala Ser Asp 370 375 3g0 Gly Lys Thr Tyr Asp Ala Tyr Ile Leu Tyr Pro Lys Thr Leu Gly Glu Gly Ser Phe Ser Asp Leu Asp Thr Phe Val Phe Lys Leu Leu Pro Glu Val Leu Glu Gly Gln Phe Gly Tyr Lys Leu Phe Ile Tyr Gly Arg Asp Asp Tyr Val Gly Glu Asp Thr Ile Glu Val Thr Asn Glu Asn Val Lys Lys Ser Arg Arg Leu Ile Ile Ile Leu Val Arg Asp Met Gly Gly Phe Ser Trp Leu Gly Gln Ser Ser Glu Glu Gln Ile Ala Ile Tyr Asn Ala Leu Ile Gln Glu Gly Ile Lys Ile Val Leu Leu Glu Leu Glu Lys Ile Gln Asp Tyr Glu Lys Met Pro Asp Ser Ile Gln Phe Ile Lys Gln Lys His Gly Val Ile Cys Trp Ser Gly Asp Phe Gln Glu Arg Pro Gln Ser Ala Lys Thr Arg Phe Trp Lys Asn Leu Arg Tyr Gln Met Pro Ala Gln Arg Arg Ser Pro Leu Ser Lys His Arg Leu Leu Thr Leu Asp Pro Val Arg Asp Thr Lys Glu Lys Leu Pro Ala Ala Thr His Leu Pro Leu Gly (2) INFORMATION FOR SEQ ID N0:31:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 569 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: not relevant (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID N0:31:
Met Lys Val Leu Leu Arg Leu Ile Cys Phe Ile Ala Leu Leu Ile Ser Ser Leu Glu Ala Asp Lys Cys Lys Glu Arg Glu Glu Lys Ile Ile Leu Val Ser Ser Ala Asn Glu Ile Asp Val Arg Pro Cys Pro Leu Asn Pro Asn Glu His Lys Gly Thr Ile Thr Trp Tyr Lys Asp Asp Ser Lys Thr Pro Val Ser Thr Glu Gln Ala Ser Arg Ile His Gln His Lys Glu Lys Leu Trp Phe Val Pro Ala Lys Val Glu Asp Ser Gly His Tyr Tyr Cys Val Val Arg Asn Ser Ser Tyr Cys Leu Arg Ile Lys Ile Ser Ala Lys Phe Val Glu Asn Glu Pro Asn Leu Cys Tyr Asn Ala Gln Ala Ile Phe Lys Gln Lys Leu Pro Val Ala Gly Asp Gly Gly Leu Val Cys Pro Tyr Met Glu Phe Phe Lys Asn Glu Asn Asn Glu Leu Pro Lys Leu Gln Trp Tyr Lys Asp Cys Lys Pro Leu Leu Leu Asp Asn Ile His Phe Ser Gly Val Lys Asp Arg Leu Ile Val Met Asn Val Ala Glu Lys His Arg Gly Asn Tyr Thr Cys His Ala Ser Tyr Thr Tyr Leu Gly Lys Gln Tyr Pro Ile Thr Arg Val Ile Glu Phe Ile Thr Leu Glu Glu Asn Lys Pro Thr Arg Pro Val Ile Val Ser Pro Ala Asn Glu Thr Met Glu Val Asp Leu Gly Ser Gln Ile Gln Leu Ile Cys Asn Val Thr Gly Gln Leu Ser Asp Ile Ala Tyr Trp Lys Trp Asn Gly Ser Val Ile Asp Glu Asp Asp~Pro Val Leu Gly Glu Asp Tyr Tyr Ser Val Glu Asn Pro Ala Asn Lys Arg Arg Ser Thr Leu Ile Thr Val Leu Asn Ile Ser Glu Ile Glu Ser Arg Phe Tyr Lys His Pro Phe Thr Cys Phe Ala Lys Asn Thr His Gly Ile Asp Ala Ala Tyr Ile Gln Leu Ile Tyr Pro Val Thr Asn ~h~ Gln Lys His Met Ile Gly Ile Cys Val Thr Leu Thr Val Ile Ile Val Cys Ser Val Phe Ile Tyr Lys Ile Phe Lys Ile Asp Ile Val Leu Trp Tyr Arg Asp Ser Cys Tyr Asp Phe Leu Pro Ile Lys Ala Ser Asp Gly Lys Thr Tyr Asp Ala Tyr Ile Leu Tyr Pro Lys Thr Val Gly Glu Gly Ser Thr Ser Asp Cys Asp Ile Phe Val Phe Lys Val Leu Pro Glu Val Leu Glu Lys Gln Cys Gly Tyr Lys Leu Phe Ile Tyr Gly Arg Asp Asp Tyr Val Gly Glu Asp Ile Val Glu Val Ile Asn Glu Asn Val Lys Lys Ser Arg Arg Leu Ile Ile Ile Leu Val Arg Glu Thr Ser Gly Phe Ser Trp Leu Gly Gly Ser Ser Glu Glu Gln Ile Ala Met Tyr Asn Ala Leu Val Gln Asp Gly Ile Lys Val Val Leu~Leu Glu Leu Glu Lys Ile Gln Asp Tyr Glu Lys Met Pro Glu Ser Ile Lys Phe Ile Lys Gln Lys His Gly Ala Ile Arg Trp Ser Gly Asp Phe Thr Gln Gly Pro Gln Ser Ala Lys Thr Arg Phe Trp Lys Asn Val Arg Tyr His Met Pro Val Gln Arg Arg Ser Pro Ser Ser Lys His Gln Leu Leu Ser Pro Ala Thr Lys Glu Lys Leu Gln Arg Glu Ala His Val Pro Leu Gly (2) INFORMATION FOR SEQ ID N0:32:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: S55 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: not relevant (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID N0:32:
Met His Lys Met Thr Ser Thr Phe Leu Leu Ile Gly His Leu Ile Leu Leu Ile Pro Leu Phe Ser Ala Glu Glu Cys Val Ile Cys Asn Tyr Phe Val Leu Val Gly Glu Pro Thr Ala Ile Ser Cys Pro Val Ile Thr Leu Pro Met Leu His Ser Asp Tyr Asn Leu Thr Trp Tyr Arg Asn Qly Ser Asn Met Pro Ile Thr Thr Glu Arg Arg Ala Arg Ile His Gln Arg Lys 65 70 75 g0 Gly Leu Leu Trp Phe Ile Pro Ala Ala Leu Glu Asp Ser Gly Leu Tyr Glu Cys Glu Val Arg Ser Leu Asn Arg Ser Lys Gln Lys Ile Ile Asn Leu Lys Val Phe Lys Asn Asp Asn Gly Leu Cys Phe Asn Gly Glu Met Lys Tyr Asp Gln Ile Val Lys Ser Ala Asn Ala Gly Lys Ile Ile Cys Pro Asp Leu Glu Asn Phe Lys Asp Glu Asp Asn Ile Asn Pro Glu Ile His Trp Tyr Lys Glu Cys Lys Ser Gly Phe Leu Glu Asp Lys Arg Leu Val Leu Ala Glu Gly Glu Asn Ala Ile Leu Ile Leu Asn Val Thr Ile Gln Asp Lys Gly Asn Tyr Thr Cys Arg Met Val Tyr Thr Tyr Met Gly Lys Gln Tyr Asn Val Ser Arg Thr Met Asn Leu Glu Val Lys Glu Ser Pro Leu Lys Met Arg Pro Glu Phe Ile Tyr Pro Asn Asn Asn Thr Ile Glu Val Glu Leu Gly Ser His Val Val Met Glu Cys Asn Val Ser Ser Gly Val Tyr Gly Leu Leu Pro Tyr Trp Gln Val Asn Asp Glu Asp Val Asp Ser Phe Asp Ser Thr Tyr Arg Glu Gln Phe Tyr Glu Glu Gly Met Pro His Gly Ile Ala Val Ser Gly Thr Lys Phe Asn Ile Ser Glu Val Lys Leu Lys Asp Tyr Ala Tyr Lys Phe Phe Cys His Phe Ile Tyr Asp Ser Gln Glu Phe Thr Ser Tyr Ile Lys Leu Glu His Pro Val Gln Asn Ile Arg Gly Tyr Leu Ile Gly Gly Gly Ile Ser Leu Ile Phe Leu Leu Phe Leu Ile Leu Ile Val Tyr Lys Ile Phe Lys Ile Asp Ile Val Leu Trp Tyr Arg Ser Ser Cys His Pro Leu Leu Gly Lys Lys Val Ser Asp Gly Lys Ile Tyr Asp Ala Tyr Val Leu Tyr Pro Lys Asn Arg Glu Ser 385 390 395 , 400 Cys Leu Tyr Ser Ser Asp Ile Phe Ala Leu Lys Ile Leu Pro Glu Val Leu Glu Arg Gln Cys Gly Tyr Asn Leu Phe Ile Phe Gly Arg Asn Asp Leu Ala Gly Glu Ala Val Ile Asp Val Thr Asp Glu Lys Ile His Gln Ser Arg Arg Val Ile Ile Ile Leu Val Pro Glu Pro Ser Cys Tyr Gly Ile Leu Glu Asp Ala Ser Glu Lys His Leu Ala Val Tyr Asn Ala Leu Ile Gln Asp Gly Ile Lys Ile Ile Leu Ile Glu Leu Glu Lys Ile Glu Asp Tyr Ala Asn Met Pro Glu Ser Ile Lys Tyr Val Lys Gln Lys Tyr Gly Ala Ile Arg Trp Thr Gly Asp Phe Ser Glu Arg Ser His Ser Ala Ser Thr Arg Phe Trp Lys Lys Val Arg Tyr His Met Pro Ser Arg Lys His Gly Ser Ser Ser Gly Phe His Leu Ser Ser (2) INFORMATION FOR SEQ ID N0:33:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 802 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: not relevant (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID N0:33:
Met Arg Leu Leu Leu Ala Leu Leu Gly Val Leu Leu Ser Val Pro Gly Pro Pro Val Leu Ser Leu Glu Ala Ser Glu Glu Val Glu Leu Glu Pro Cys Leu Ala Pro Ser Leu Glu Gln Gln Glu Gln Glu Leu Thr Val Ala Leu Gly Gln Pro Val Arg Leu Cys Cys Gly Arg Ala Glu Ark Gly Gly His Trp Tyr Lys Glu Gly Ser Arg Leu Ala Pro Ala Gly Arg Val Arg 65 70 75 g0 Gly Trp Arg Gly Arg Leu Glu Ile Ala Ser Phe Leu Pro Glu Asp Ala Gly Arg Tyr Leu Cys Leu Ala Arg Gly Ser Met Ile Val Leu Gln Asn Leu Thr Leu Ile Thr Gly Asp Ser Leu Thr Ser Ser Asn Asp Asp Glu Asp Pro Lys Ser His Arg Asp Pro Ser Asn Arg His Ser Tyr Pro Gln Gin Ala Pro Tyr Trp Thr His Pro Gln Arg Met Glu Lys Lys Leu His Ala Val Pro Ala Gly Asn Thr Val Lys Phe Arg Cys Pro Ala Ala Gly Asn Pro Thr Pro Thr Ile Arg Trp Leu Lys Asp Gly Gln Ala Phe His Gly Glu Asn Arg Ile Gly Gly Ile Arg Leu Arg His Gln His Trp Ser Leu Val Met Glu Ser Val Val Pro Ser Asp Arg Gly Thr Tyr Thr Cys Leu Val Glu Asn Ala Val Gly Ser Ile Arg Tyr Asn Tyr Leu Leu Asp Val Leu Glu Arg Ser Pro His Arg Pro Ile Leu Gln Ala Gly Leu Pro Ala Asn Thr Thr Ala Val Val Gly Ser Asp Val Glu Leu Leu Cys Lys Val Tyr Ser Asp Ala Gln Pro His Ile Gln Trp Leu Lys His Ile Val Ile Asn Gly Ser Ser Phe Gly Ala Val Gly Phe Pro Tyr Val Gln Val Leu Lys Thr Ala Asp Ile Asn Ser Ser Glu Val Glu Val Leu Tyr Leu Arg Asn Val Ser Ala Glu Asp Ala Gly Glu Tyr Thr Cys Leu Ala Gly 325 330 335 .
Asn Ser Ile Gly Leu Ser Tyr Gln Ser Ala Trp Leu Thr Val Leu Pro Glu Glu Asp Pro Thr Trp Thr Ala Ala Ala Pro Glu Ala Arg Tyr Thr Asp Ile Ile Leu Tyr Ala Ser Gly Ser Leu Ala Leu Ala Val Leu Leu Leu Leu Ala Gly Leu Tyr Arg Gly Gln Ala Leu His Gly Arg His Pro 385 390 395 ~ ~ 400 Arg Pro Pro Ala Thr Val Gln Lys Leu Ser Arg Phe Pro Leu Ala Arg Gln Phe Ser Leu Glu Ser Gly Ser Ser Gly Lys Ser Ser Ser Ser Leu Val Arg Gly Val Arg Leu Ser Ser Ser Gly Pro Ala Leu Leu Ala Gly Leu Val Ser Leu Asp Leu Pro Leu Asp Pro Leu Trp Glu Phe Pro Arg Asp Arg Leu Val Leu Gly Lys Pro Leu Gly Glu Gly Cys Phe Gly Gln Val Val Arg Ala Glu Ala Phe Gly Met Asp Pro Ala Arg Pro Asp Gln Ala Ser Thr Val Ala Val Lys Met Leu Lys Asp Asn Ala Ser Asp Lys Asp Leu Ala Asp Leu Val Ser Glu Met Glu Val Met Lys Leu Ile Gly 515 , 520 525 Arg His Lys Asn Ile Ile Asn Leu Leu Gly Val Cys Thr Gln Glu Gly Pro Leu Tyr Val Ile Val Glu Cys Ala Ala Lys Gly Asn Leu Arg Glu Phe Leu Arg Ala Arg Arg Pro Pro Gly Pro Asp Leu Ser Pro Asp Gly Pro Arg Ser Ser Glu Gly Pro Leu Ser Phe Pro Val Leu Val Ser Cys Ala Tyr Gln Val Ala Arg Gly Met Gln Tyr Leu Glu Ser Arg Lys Cys Ile His Arg Asp Leu Ala Ala Arg Asn Val Leu Val Thr Glu Asp Asn Val Met Lys Ile Ala Asp Phe Gly Leu Ala Arg Gly Val His His Ile Asp Tyr Tyr Lys Lys Thr Ser Asn Gly Arg Leu Pro Val Lys Trp Met Ala Pro Glu Ala Leu Phe Asp Arg Val Tyr Thr His Gln Ser Asp Val Trp Ser Phe Gly Ile Leu Leu Trp Glu Ile Phe Thr Leu Gly Gly Ser WO 99/19480 PC'fNS98/20939 Pro Tyr Pro Gly Ile Pro Val Glu Glu Leu Phe Ser Leu Leu Arg Glu Gly His Arg Met Asp Arg Pro Pro His Cys Pro Pro Glu Leu Tyr Gly Leu Met Arg Glu Cys Trp His Ala Ala Pro Ser Gln Arg Pro Thr Phe Lys Gln Leu Val Glu Ala Leu Asp Lys Val. Leu Leu Ala Val Ser Glu Glu Tyr Leu Asp Leu Arg Leu Thr Phe Gly Pro Tyr Ser Pro Ser Gly Gly Asp Ala Ser Ser Thr Cys Ser Ser Ser Asp Ser Val Phe Ser His Asp Pro Leu Pro Leu Gly Ser Ser Ser Phe Pro Phe Gly Ser Gly Val Gln Thr

Claims

WHAT IS CLAIMED IS:
1. An isolated or recombinant IL-1RD9 polypeptide:
a) consisting of SEQ ID NO: 6, 8, 10, 12, 14, or 16;
b) encoded by a polynucleotide comprising the open reading frame of SEQ ID NO: 5, 7, 9, 11, 13; or 15; or c) encoded by a naturally occurring allelic variant of a polynucleotide comprising the open reading frame of SEQ ID NO: 5, 7, 9, 11, 13, or 15.
2. The polypeptide of claim 1, encoded by a naturally occurring allelic variant of a polynucleotide comprising the open reading frame of SEQ ID NO: 5, 7, 9, 11, 13, or 15.
3. An isolated or recombinant Il-1RD9 polypeptide which:
a) has an apparent molecular weight 68.3 kD as determined by SDS/polyacrylamide gel electrophoresis;
b) has as estimated pI of 9.04; and c) is found on T cells; and wherein said polypeptide has at least one of the following properties:
i) is a heterodimer;
iii) is an IL-1 .alpha. subunit type, or iii) when brought into contact with IL-1RD5 and IL-1.alpha., for a sufficient time, forms a functional high affinity receptor complex that activates an NF~B transcription factor reporter construct.
4. An isolated or recombinant polypeptide comprising a segment of contiguous amino acid residues selected from the following group:
a) 15 contiguous amino acid residues of said polypeptide of claim 2;

b) 20 contiguous amino acid residues of said polypeptide of claim 2;
c) 25 contiguous amino acid residues of said polypeptide of claim 2;
d) 30 contiguous amino acid residues of said polypeptide of claim 2;
e) 35 contiguous amino acid residues of said polypeptide of claim 2; or f) 40 contiguous amino acid residues of said polypeptide of claim 2.
5. The polypeptide of claim 1 which is immunogenic.
6. An isolated or recombinant polypeptide comprising an immunogenic peptide of said polypeptide of claim 3.
7. An isolated or recombinant polypeptide comprising an immunogenic polypeptide of claim 4.
8. A fusion protein comprising said polypeptide of claim 4 and:
a) a detection or purification tag selected from the group consisting of a FLAG, His6, and immunoglobulin peptide;
b) a carrier protein selected from the group consisting of keyhole limpet hemocyanin, bovine serum albumin, and tetanus toxoid; or c) another peptide selected from the group consisting of luciferase, bacterial .beta.-galactosidase, trpE, protein A, .beta.-lactamase, alpha amylase, alcohol dehydrogenase, and yeast alpha mating factor.
9. A fusion protein comprising said polypeptide of claim 5 and:
a) a detection or purification tag selected from the group consisting of a FLAG, His6, and immunoglobulin peptide;

b) a carrier protein selected from the group consisting of keyhole limpet hemocyanin, bovine serum albumin, the tetanus toxoid; or c) another peptide selected from the group consisting of luciferase, bacterial .beta.-galactosidase, trpE, protein A, .beta.-lactamase, alpha amylase, alcohol dehydrogenase, and yeast alpha mating factor.
10. A composition comprising said polypeptide of claim 1, that is:
a) in a pharmaceutically acceptable carrier;
b) in a sterile composition;
c) in a buffered solution; or d) in an aqueous suspension.
11. A composition comprising said polypeptide of claim 4, that is:
a) in a pharmaceutically acceptable carrier;
b) in a sterile composition;
c) in a buffered solution; or d) in an aqueous suspension.
12. A polypeptide of claim 4, that is:
a) denatured;
b) immunopurified;
c) attached to a solid substrate;
d) detestably labeled; or e) chemically synthesized.
13. A polypeptide of claim 5, that is:
a) denatured;
b) immunopurified;
c) attached to a solid substrate;
d) detestably labeled; or e) chemically synthesized.
14. A kit comprising said polypeptide of claim 1, and:
a) a compartment comprising said protein; or b) instructions for use or disposal of reagents in said kit.
15. A kit comprising said polypeptide of claim 4, and:
a) a compartment comprising said protein; or b) instructions for use or disposal of reagents in said kit.
16. A method of raising an antibody, comprising immunizing an animal with a polypeptide of claim 5.
17. A method of producing an antibody:antigen complex, comprising contacting a polypeptide of claim 5 with an antibody which specifically binds said polypeptide, thereby forming said complex.
18. A composition of matter selected from the group consisting of:
a) a substantially pure or recombinant IL-1RD8 polypeptide exhibiting identity over a length of at least about 12 amino acids to SEQ ID NO:
4;
b) a natural sequence IL-1RD8 comprising SEQ ID NO:
4;
c) a fusion polypeptide comprising IL-1RD8 sequence;
d) a substantially pure or recombinant IL-1RD10 polypeptide exhibiting identity over a length of at least about 12 amino acids to SEQ ID NO:
20;
e) a natural sequence IL-1RD10 comprising SEQ ID
NO: 20; and f) a fusion protein comprising IL-1RD10 sequence.
19. A substantially pure or isolated polypeptide comprising a segment exhibiting sequence identity to a corresponding portion of an:
a) IL-1RD8 of claim 18, wherein:

i) said polypeptide further exhibits identity to a distinct segment of 9 amino acids;
ii) said length of identity is at least l7 amino acids;
iii) said length of identity is at least about 25 amino acids; or b) IL-1RD10 of claim 18, wherein:
i) said polypeptide further exhibits identity to a distinct segment of 9 amino acids;
ii) said length of identity is at least 17 amino acids;
iii) said length of identity is at least about 25 amino acids.
20. The composition of matter of claim 18, wherein said:
a) IL-1RD8 comprises a mature sequence of SEQ ID NO
2 or 4;
b) IL-1RD10 comprises a mature sequence of Seq ID
NO: 18 or 20; or c) polypeptide:
i) is from a warm blooded animal selected from a primate, such as a human;
ii) comprises at least one polypeptide segment of SEQ ID NO: 4 or 20;
iii) exhibits a plurality of portions exhibiting said identity;
iv) is a natural allelic variant of a primate or rodent IL-1RD8 or primate IL-1RD10;
v) has a length at least about 30 amino acids;
vi) exhibits at least two non-overlapping epitopes which are specific for a primate or rodent IL-1RD8 or primate IL-1RD10;
vii) exhibits a sequence identity at least about 90% over a length of at least about 20 amino acids to a primate IL-1RD8 or IL-1RD10;
viii) has a molecular weight of at least 100 kD with natural glycosylation;
ix) is a synthetic polypeptide;

x) is attached to a solid substrate;
xi) is conjugated to another chemical moiety;
xii) is a 5-fold or less substitution from natural sequence; or xiii) is a deletion or insertion variant from a natural sequence.
21. A composition comprising:
a) a sterile IL-1RD8 polypeptide of claim 18;
b) said IL-1RD8 protein or peptide of claim 18 and a carrier, wherein said carrier is:
i) an aqueous compound, including water, saline, and/or buffer; and/or ii) formulated for oral, rectal, nasal, topical, or parenteral administration;
c) a sterile IL-1RD10 polypeptide of claim 18; or d) said IL-1RD10 polypeptide of claim 18 and a carrier, wherein said carrier is:
i) an aqueous compound, including water, saline, and/or buffer; and/or ii) formulated for oral, rectal, nasal, topical, or parenteral administration.
21. A fusion protein of claim 18, comprising:
a) mature protein sequence of SEQ ID NO: 2, 4, 18 or 20;
b) a detection or purification tag, including a FLAG, His6, or Ig sequence; or c) sequence of another receptor protein.
22. A kit comprising a polypeptide of claim 18, and:
a) a compartment comprising said polypeptide;
and/or b) instructions for use or disposal of reagents in said kit.
23. A binding compound comprising an antigen binding site from an antibody, which specifically binds to a natural:

A) IL-1RD8 protein of claim 18, wherein:
a) said protein is a primate or rodent protein;
b) said binding compound is an Fv, Fab, or Fab2 fragment;
c) said binding compound is conjugated to another chemical moiety; or d) said antibody:
i) is raised against a peptide sequence of a mature polypeptide of Seq ID NO
2 or 4;
ii) is raised against a mature primate or rodent IL-1RD8;
iii) is raised to a purified human IL-1RD8;
iv) is raised to a purified mouse IL-1RD8;
v) is immunoselected;
vi) is a polyclonal antibody;
vii) binds to a denatured IL-1RD8 ;
viii) exhibits a Kd to antigen of at least 30 µM;
ix) is attached to a solid substrate, including a bead or plastic membrane;
x) is in a sterile composition; or xi) is detectably labeled, including a radioactive or fluorescent label; or B) IL-1RD10 polypeptide of claim 18, wherein:
a) said polypeptide is a primate polypeptide;
b) said binding compound is an Fv, Fab, or Fab2 fragment;
c) said binding compound is conjugated to another chemical moiety; or d) said antibody:
i) is raised against a peptide sequence of a mature polypeptide of SEQ ID NO:
18 or 20;
ii) is raised against a mature primate IL-1RD10;

iii) is raised to a purified human IL-1RD10;
iv) is immunoselected;
v) is a polyclonal antibody;
vi) binds to a denatured IL-1RD10;
vii) exhibits a Kd to antigen of at least 30 µM;
viii) is attached to a solid substrate, including a bead or plastic membrane;
ix) is in a sterile composition; or x) is detectably labeled, including a radioactive or fluorescent label 24. A kit comprising said binding compound of claim 25, and:
a) a compartment comprising said binding compound;
and/or b) instructions for use or disposal of reagents in said kit.

26. A method of:
A) making an antibody of claim 23, comprising immunizing an immune system with an immunogenic amount of:
a) a primate IL-1RD8 polypeptide;
b) a primate IL-1RD10 polypeptide;or thereby causing said antibody to be produced;
or B) producing an antigen: antibody complex, comprising contacting:
a) a primate IL-1RD8 polypeptide with an antibody of claim 23A; or b) a primate IL-1RD10 polypeptide with an antibody of claim 23B;
thereby allowing said complex to form.

27. A composition comprising:
a) a sterile binding compound of claim 23, or b) said binding compound of claim 23 and a carrier, wherein said carrier is:
i) an aqueous compound, including water, saline, and/or buffer; and/or ii) formulated for oral, rectal, nasal, topical, or parenteral administration.

28. An isolated or recombinant nucleic acid encoding a protein or peptide or fusion protein of claim 18, wherein:
a) said IL-1RD8 or IL-1RD10 is from a mammal; or b) said nucleic acid:
i) encodes an antigenic polypeptide sequence of SEQ ID NO: 2, 4, 18 or 20;
ii) encodes a plurality of antigenic polypeptide sequences of SEQ ID NO: 2, 4, 18 or 20;
iii) exhibits identity to a natural cDNA
encoding said segment;
iv) is an expression vector;
v) further comprises an origin of replication;
vi) is from a natural source;
vii) comprises a detectable label;
viii) comprises synthetic nucleotide sequence;
ix) is less than 6 kb, preferably less than 3 kb;
x) is from a mammal, including a primate, such as a human;
xi) comprises a natural full length coding sequence;
xii) is a hybridization probe for a gene encoding said IL-1RD8 or IL-1RD10;
xiii) comprises a plurality of nonoverlapping segments of at least 15 nucleotides from SEQ ID NO: 1, 3, 17 or 19; or xiv) is a PCR primer, PCR product, or mutagenesis primer.

29. A cell transfected or transformed with a recombinant nucleic acid of claim 28.

30. The cell of claim 29, wherein said cell is:
a) a prokaryotic cell;
b) a eukaryotic cell;
c) a bacterial cell;
d) a yeast cell;
e) an insect cell;
f) a mammalian cell;
g) a mouse cell;
h) a primate cell; or i) a human cell.

31. A kit comprising said nucleic acid of claim 28, and:
a) a compartment comprising said nucleic acid;
b) a compartment further comprising a primate or rodent IL-1RD8 or primate IL-1RD10 polypeptide;
and/or b) instructions for use or disposal of reagents in said kit.

32. A method of:
A) making a polypeptide, comprising expressing said nucleic acid of claim 28, thereby producing said polypeptide; or B) making a duplex nucleic acid, comprising contacting said nucleic acid of claim 28 with a hybridizing nucleic acid, thereby allowing said duplex to form.

33. A nucleic acid which:
a) hybridizes under wash conditions of 40° C and less than 2M salt to SEQ ID NO: 3 or 19; or b) exhibits identity over a stretch of at least about 30 nucleotides to a primate IL-1RD8 or IL-1RD10.

34. The nucleic acid of claim 33, wherein:

84~

a) said wash conditions are at 55° C and/or 500 mM
salt; or b) said stretch is at least 55 nucleotides.

35. The nucleic acid of claim 34, wherein:
a) said wash conditions are at 65° C and/or 150 mM
salt; or b) said stretch is at least 75 nucleotides.

36. A method of modulating physiology or development of a cell or tissue culture cells comprising contacting said cell with an agonist or antagonist of a primate IL-1RD8 or IL-1RD10.

37. The method of claim 36, wherein said cell is transformed with a nucleic acid encoding either an IL-1RD8 or IL-1RD10, and another IL-1R.