CA2265470A1 - Biochemical markers of the human endometrium - Google Patents

Abstract

Assay methods are provided for detection or quantitation of any of several proteins which are specifically produced in the endometrium in association with hyperplasia, adenocarcinoma or the proliferative phase of the endometrium. The relevant proteins have been identified by 2D gel electrophoresis with subsequent sequence identification by mass spectroscopic finger printing of tryptic digests.

Description

?101520253035CA 02265470 1999-03-05W0 93/10291 PCT/GB97/02394-1-BIOCHEMICAL MARKERS OF THE HUMAN ENDOMETRIUMThe endometrium is the mucous lining of the uterinecavity. During the menstrual cycle, the endometrium is theorgan in the body that shows the greatest changes under theinfluence of the sex hormones, cestradiol and progesterone.In the oestrogen dominated phase the endometriumproliferates until progesterone from the corpus luteumtransforms the oestrogen—primed proliferative endometrium toa secretory phase endometrium. In due course this isfollowed by shedding of the fully transformed endometriumduring the menstruation, and a new cycle will begin.Persistent unbalanced oestrogen stimulation either dueto increased endogenous production of oestrogens, orreplacement therapy in which oestrogens are given alone, isassociated with increased risk of developing endometrialhyperplasia and subsequently endometrial adenocarcinoma.Histologically, these pathological conditions arecharacterised by increased thickness of the endometrium andirregular pattern of the endometrial glandular cells.Endometrial adenocarcinoma is a life threateningcondition.At present the endometrial status is assessed byhistological and biochemical analysis of endometrialbiopsies. This is time-consuming, expensive and causesdiscomfort for the woman. It would be highly desirable toidentify biochemical markers which could be measured in bodyfluids reflecting the endometrial status, obviating the needfor endometrial biopsies. The detection of such markers inhistological samples would also however be advantageous asan additional method of recognising the histological statusof such samples.thatproduced in the endometrium in increased amounts associatedWe have now discovered certain proteins arewith hyperplasia and that certain proteins are produced inincreased amounts associated with adenocarcinoma. These twogroups of proteins overlap somewhat. The present inventionrelates in a first aspect to such proteins and to theirdiagnostic uses.?l01520253035CA 02265470 1999-03-05WO 98/1029].2.Unless otherwise indicated, references to the proteinsherein include references to modified forms of the proteinsand derivatives of the proteins, including but notrestricted to glycosylated, phosphorylated, acetylated,methylated or lipidated forms thereof.Thus the invention provides a method of characterisinga biological sample comprising detecting or quantitatingtherein one or more proteins produced by the endometrium inincreased amounts in hyperplasia or in adenocarcinoma asshown by 2D gel electrophoresis comparison of cell lysatesof endometrial biopsies from normal endometrium andendometrium showing hyperplasia or adenocarcinoma,to thequantitating a fragment or breakdown product thereof,excludingvariations due menstrual cycle, or detecting oror anucleic acid coding therefor, or an antibody thereto.Thebiologicalinvention includes a method of characterising asample comprising detecting or quantitatingtherein one or more proteins produced by the endometrium inincreased amounts in. hyperplasia. or i11 adenocarcinoma andthe following combinations ofcharacterised by one ofmolecular weight and p1 values:hyperplasiapl MW kDa91906467664641351310114514448\.0CD\lU'|O1U'|U1U'IO\O\O\O\O\O\U'lt\)v¥>(IJO‘\bJU1\'lmL\JO\\.00‘\\)PCT/GB97/02394?lO1520253035CA 02265470 1999-03-05W0 98/1029] PCT/GB97/02394adenocarcinomapI MW (kDa)3210991906467666245453327103907813130667332554411143483239qooxoqoxoooooommmmmmmmmmmmmmmmmmm\1wu1\:mmo—-uwwmwmmmwwqmkvuzmioo?bbor a fragment or breakdown product thereof, or a nucleicacid coding therefor, or an antibody thereto.Said protein, fragment, breakdown product, antibody ornucleic acid may preferably be detected in a body fluidsample but may also be detailed in other forms of samplesuch as histological samples or cytological samples.?1O1520253035ICA 02265470 1999-03-05WO 98/10291 PCT/GB97/02394The invention includes an immunological binding partnerspecifically reactive with a protein as defined above with afragment or breakdown product thereof or with a nucelic acidcoding therefor.It also includes a cell line producing a Inonoclonalantibody being such an immunological binding partner.The invention includes also an assay kit for use insuch an analysis method comprising an immunological bindingpartner as described.This aspect of the invention has resulted from studiesaiming to detect endometrial proteins with increasedsynthesis in endometrial adenocarcinoma as compared to thesynthesis during the normal menstrual cycle; to detectendometrial proteins with increased synthesis in endometrialhyperplasia as compared to the synthesis during the normalmenstrual cycle; and to detect proteins showing acycle—related expression during the normal menstrual cycle.the thephase of theIn a second aspect invention relates todiscovery of markers of the "proliferative"human endometrium. A protein marker for the "secretory"phase of the endometrium has been previously described, seeUS—A-4, 489, 166.the proliferative phase although certain candidate proteinsNo similar marker has been described forwere described in Ref. 1.Under influence of thethesex hormones, oestradiol andendometrium undergoes cyclicalthethe intervalprogesterone, humanvariation with an oestrogen—dominated phase, i.e.proliferative phase, an ovulation phase, i.e.i.e. the secretoryshed, thevariation of thea progesterone-dominated. phase,theTheseen inphase,phase, and finally endometrium is i.e.menstrual phase. same cyclicalpostmenopausal women receivingThehighlynot only on account of theendometrium issequentially combined hormone replacement therapy.demand for endometrial status assessment hasincreased in the latest decade,extensive research into fertility, but also in order to?10152025CA 02265470 1999-03-05W0 98/ 10291 PCT/GB97l02394- 5 _estimate endometrial response to the large number ofcombined oestrogens/progestogen preparations used in hormonereplacement therapy. It would be highly desirable toidentify biochemical markers which could be measured in bodyfluids reflecting the endometrial status, obviating the needfor endometrial biopsies. Studies have suggested that serum(PP14) .glandular cells of the secretory phase endometrium (Ref. 3),placental protein 14 which is produced in theis a reliable marker of the secretory phase endometrium. Ithas been shown that serum PP14 strongly correlates with thethewomen receiving hormone replacement therapy (Ref. 4,5). Nothesecretory activity of endometrium in postmenopausalsimilar marker exists for proliferative phaseendometrium.We have now discovered that certain proteinstheareproduced in endometrium in increased amounts inproliferative phase endometrium as compared to secretoryphase endometrium.According to this aspect of the invention there is nowprovided a method of characterising a biological samplecomprising detecting or quantitating therein one or moreproteins produced. by the endometriunm in increased. amountsduring the proliferative phase of the endometrium as showncomparison of cell lysates ofin 2D gel elctrophoresisendometrial biopsies from normal endometrium in itsproliferative and secretory phases and characterised by oneof the following combinations of molecular weight and pIvalues:—?1015ICA 02265470 1999-03-05W0 98/10291 PCT/GB97/02394.5.p1 MW(kDa)6.9 865.4 345.6 675.3 236.8 528.7 478.2 1386.5 1247.7 1197.8 1198.1 667.1 596.8 667.9 487.7 316.8 297.2 708.0 1196.7 62or a fragment or breakdown product thereof, or a nucleicacid coding therefor or an antibody thereto.Such a method may preferably be for detecting the phaseof the endometrium.The preferred features of the first aspect of theinvention apply also to this second aspect.This of thedetermining the proliferative/secretory phase status of thetheaspect invention includes a method ofqualitativeendometrium quantitative orcomprisingmeasurement in a sample of any one or more of the proteinsdefined above or a breakdown product or fragment thereof.It also includes an immunological binding partner for any ofthe said proteins, breakdown products or fragments or a cellline producing such a binding partner.?101520253035CA 02265470 1999-03-05W0 98/10291 PCTIGB97/02394_ 7 _Whilst the sequences and properties of proteinsdiscussed above relate to human proteins, the assayprocedures of the invention may be practised on samplesEspecially in this context,should beinclude proteins having a degree of homology of at least 60%arising from other species.references to proteins herein understood towith the given amino acid sequences irrespective of anymodifications of said amino acids. When determininghomology, modified amino acids such as phosphorylated,acetylated, amidated, methylated, glycosylated or lipidatedderivatives of an amino acid should thus be considered to bethe same as the amino acid without any such modification.Such peptides may be derived from similar proteins frome.g. rabbit,guinea pig, pig, or cow or may be entirely or predominantlyother species, other mammals such as mouse,of synthetic origin.The degree of homology may be advantageously be atleast 65%,is advantageous that the degree of homology is even higheror at least 70%. Under certain circumstances, itsuch as at least 80% or at least 90%. Other DNA sequenceswhich encode substantially the same amino acid sequence as agene encoding a marker protein, i.e. a marker gene, may beused in the practice of the present invention. Theseinclude, but are not limited to, allelic genes andhomologous genes from other species.Nucleic acid fragments comprising a nucleotide sequencewhich codes for a protein described above or a peptidederived from it as well as nucleic acid fragments whichwiththereof under stringent hybridisation conditions,acid fragments or a parte.g. 5 mMmonovalent ions (O.lXSSC), neutral pH and 65°C are importanthybridise these nucleicaspects of the invention. The term "highly stringent", whenused in conjunction with hybrisidation conditions, is asdefined in the art, i.e. 5—l0°C under the melting point Tmcf, 1989, 11.49.By the term "nucleic acid" is meant a polynucleotide ofhigh molecular weight which can occur as either DNA or RNAand may be either single-stranded or double-stranded.Sambrook et al, pages 11.45 -?l01520253035ICA 02265470 1999-03-05W0 98/10291 PCT/GB97/02394-8-the amino acid sequences of the proteins ofutility in the present invention are known,Onceit is possibleto synthesise DNA or RNA probes which may be used for:i) direct detection of DNA and RNA expressing saidfrozen tissue sectionproteins on a fixed ore.g.immunofluorescent techniques;using, chromogenous, chemiluminescent orii) polymerase chain reaction (PCR) or otheramplification techniques; andiii) locating the part or all of the gene, isogene,pseudogene or other related genes either in cDNAlibraries, genomic libraries or other collectionsof genetic material from either the host or otheranimals, including man.In another aspect, the invention relates to a bindingmeans which specifically binds to a relevant protein orpeptide or nucleic acid fragment as described above. Inparticular, the invention relates to an antibody whichspecifically binds to a relevant protein or peptide or anantigen—binding fragment thereof, i.e. a polyclonalantibody, a monoclonal antibody, chimeric antibody, singlechain antibody fragment, Fab and Fab’ fragments, and an Fabexpression library.It is contemplated that both monoclonal and polyclonalantibodies will be useful in providing the basis for one ormore assays to detect relevant peptides and proteins.Antibodies which are directed against epitopes that arespecific for the proteins will be most useful as crossreaction will be minimised therewith.Based upon the identification of relevant proteinsdescribed above, assay methods and kits may be producedaccording to standard methodology. Thus, the proteins maybe obtained in purified form, either by extraction fromtissues or by synthesis, and antibodies may be raisedthereto or to characterising peptide sequences thereof.Standard assay formats employing such antibodies may beutilised according to the invention.?l0l520253035CA 02265470 1999-03-05W0 98/ 10291 PCT/GB97/02394-9-Preferred immunoassays are contemplated as includingvarious types of enzyme linked immunoassays (ELISA),immunoblot techniques, and the like, known in the art.However, it is readily appreciated that utility is notlimited to such assays, and useful embodiments includingRIAs and other non—enzyme linked antibody binding assays orprocedures. The proteins themselves or peptides derivedfrom the protein sequences may be used in detecting auto-antibodies to such proteins.Samples of the proteins described above have beensubjected to trypsin digestion and the molecular weight ofthe resulting fragments has been determined by massspectrometry. This provides a "fingerprint" of the proteinwhich can be matched to date in established data basesfield.has enabled us to identify certain of the proteins as beingavailable to those working in this This procedurepreviously known in other contexts. No matches have beenfound for certain others, indicating that they have notpreviously been known.The invention will be illustrated and explained furtherby the which thefollows:-following description in Figures asFigure 1: Fluorograph of a two—dimensional gel electro-phoresis of [”S]methionine labelled endometrialproteins separated in the first dimension by iso-(IEF; pl 3.5-7) thesodium dodecyl sulphateelectric focusing and insecond dimension bypolyacrylamide gel electrophoresis. The locationsof the withhyperplasia are indicated.spots increased synthesis intwo—dimensional gel electro—labelledproteins separated in the first dimension by non-Figure 2: Fluorograph of aphoresis of [”S]methionine endometrialequilibrium pH gradient gel(NEPHGE; pl 6.5-ll) and in the second dimension byelectrophoresissodium dodecyl sulphate polyacrylamide gel?101520253035W0 98/10291Figure 3:Figure 4:Figure 5:Figure 6:Figure 7:ICA 02265470 1999-03-05PCTIGB97/02394_10-electrophoresis. The locations of the spots withincreased synthesis in hyperplasia are indicated.of a gel electro-[35Fluorograph two-dimensionalof labelledproteins separated in the first dimension by iso-(IEF; pl 3.5-7) theby dodecyl sulphatepolyacrylamide gel elctrophoresis.of the withadenocarcinoma are indicated.phoresis S]methionine endometrialelectric focusing and insecond dimension sodiumThe locationsincreased synthesisspots inFluorograph of a two-dimensional gel electro-of labelledproteins separated in the first dimension by non-pH(NEPHGE; pI 6.5-ll)dodecylphoresis [”S]methionine endometrialequilibrium gradient gel electrophoresisand in the second dimension bysodium sulphatepolyacrylamide gelelectrophoresis. The locations of the spots withincreased synthesis in adenocarcinoma areindicated.Fluorograph of a two-dimensional gel electro-of labelledproteins separated in the first dimension by iso-(IEF; pI 3.5-7) theby sodium dodecyl sulphatepolyacrylamide gel elctrophoresis. Theof the with synthesisproliferative phase endometrium are indicated.phoresis f5S]methionine endometrialelectric focusing and insecond dimensionlocationsinspots increasedFluorograph of a two-dimensional gel electro-of labelledproteins separated in the first dimension by non-pH(NEPHGE; pl 6.5-11)dodecylphoresis [”S]methionine endometrialequilibrium gradient gel electrophoresisand in the second dimension bysodium sulphate polyacrylamide gelelectrophoresis. The locations of the spots withincreased synthesis in proliferative phaseendometrium are indicated.Tryptic digestion mass spectroscopic charac-teristics of I#350. The peaks marked with a star?101520253035CA 02265470 1999-03-05W0 98ll029l PCTIGB97/02394-11-are not protein identification specific butrepresents methodologically non-specific peaks.Figure 8: Tryptic digestion mass spectroscopic charac-teristics of I#687. The peaks marked with a starare not protein identification specific butrepresents methodologically non-specific peaks.massTryptic charac-teristics of N#4l4.Figure 9: digestion spectroscopicThe peaks marked with a starare not protein identification specific butrepresents methodologically non-specific peaks.Figure 10: Tryptic digestion mass spectroscopic charac-teristics of I#lO35. The peaks marked with a starare not protein identification specific butrepresents methodologically non-specific peaks.Figure 1l:Tryptic digestion mass charac-teristics of N#26.spectroscopicThe peaks marked with a starare not protein identification specific butrepresents methodologically non-specific peaks.Figure l2:Tryptic digestion mass spectroscopic charac-teristics of N#31+N#32.star are not protein identification specific butThe peaks marked with arepresents methodologically non-specific peaks.To identify proteins expressed at an increased level indiffering endometrial conditions, endometrial samples wereobtained as follows.Normal menstrual cycle samples were obtained asdescribed in Ref. 1. Endometrial biopsies were collectedregular cycling women(n=l)for‘ a variety of medicalfrom 13 pre—menopausal, (35-50 yearsold)(removal of the uterus)undergoing endometrial curettage(n=l2)not related to abnormality or malignancy of theor hysterectomyreasonsendometrium. None used hormone contraception. Forcondition samples,collected from 16 patients (41 to 79 years old)(n=9) orpathological endometrial biopsies wereundergoing(n=7)medical reasons related to abnormality or malignancy of theendometrial curettage hysterectomy forendometrium.?101520253035CA 02265470 1999-03-05wo 93/10291 PCT/GB97/02394-12-The samples were treated as described in Ref. 1.the“S-methionine for 20 hours,2D gelseparatedThemetabolicallyand total cellproteins of endometrial biopsies werelabelled withwere for electrophoresis, alysates processedtechnique in which proteins are in the firstdimension according to the isoelectric point and in thesecond dimension according to the molecular weight. It waspossible to study proteins with iso—electric points rangingfrom 3.5 to 11 and relative molecular weights ranging from10 to 300 kDa. After electrophoresis the gels were fixed andof the 2D gelelectrophoresis were subjected to quantitative analysis bytreated for fluorography. The fluorogramscomputer-aided analysis, by which the density of each spotwas quantified, the fluorogranl patterns were matched i.e.numbers were assigned to each. spot and the same spot wasgiven the same number on all the fluorograms. The density(quantity synthesis) of each spot was assessed to findproteins with increased synthesis in endometrialadenocarcinoma or hyperplasia and assessed for periodiccharacteristics during the normal menstrual cycle to findproteins with the menstrual cycle-related synthesis.Some of the menstrual cycle-related proteins soidentified have been identified by amino acid sequenceanalysis (Ref.2). Selected menstrual cycle-relatedproteins were excised from several 2D gels, concentrated by1D sodium dodecylsulphate polyacrylamide gel electro-horesis, and cleaved in situ by trypsin. The trypticfragments were extracted and separated by reverse phase highpressure liquid chromatography. Finally, the partialamino—terminal amino acid sequence of selected trypticfragments were determined for each protein. Foridentification the amino acid sequences of the trypticfragments were compared to previously reported sequences bysearching in databases.The hyperplasia and adenocarcinoma associated proteinsof thecharacterised by similar methods.present invention may be sequenced and further?l015202530CA 02265470 1999-03-05W0 98/ 10291 PCTIGB97/02394-13-Out of a total number of approximately 1,700 spots, 14spots were found to have increased synthesis in hyperplasia.The locations of these are shown in Figures 1 and 2. SomeTheTheinformation obtained from the 2D—gel electrophoresis with(pl)27 spots had increased synthesis in adenocarcinoma.locations of these are shown in Figures 3 and 4.isoelectric point and the molecularthe with increasedgiven in Table 1, theincreased synthesis in adenocarcinoma are listedrespect to the(MW) ofhyperplasia issynthesis inwithin Table 2.hyperplasiaweight spotsand spotsEight spots had increased expression in. bothand adenocarcinoma. Based on subjective evaluation,preferred subgroups of spots were selected. with increasedsynthesis in hyperplasia and in adenocarcinoma,The withincreased synthesis in hyperplasia were selected as beingrespectively. preferred subgroup of spotsthe spots showing the highest relative increase inexpression in hyperplasia as compared to the samplesobtained from women during the normal mentrual cycle andwomen with irregular proliferative phase endometrium.Similarly, the preferred. subgroup of spots with increasedsynthesis in adenocarcinoma were selected as the spotsshowing the highest relative increase in expression inadenocarcinoma as compared to the samples obtained fromwomen during the normal menstrual cycle and women withirregular proliferative phase endometrium. The preferredsubgroup of 7 spots with increased synthesis in hyperplasiais given in Table 3, and the preferred subgroup of 12 spotswith increased synthesis in adenocarcinoma is given in Table4.?CA 02265470 1999-03-05WO 98/10291 PCT/GB97/02394-14-TABLE 1Endometrial proteins with increased synthesis inhyperplasiaMatch # pI MW(kDa)I#lll 6.7 91I#l2l 6.6 90I#l58 6.9 64I#l77 6.6 67I#l9l 6.3 66I#307 6.8 46I#35O 5.7 41I#405 5.5 35I#G53 5.3 13I#892 6.6 101I#ll83 5.8 14N#l26 7.4 51N#l48 8.2 44N#4l4 9.5 48?CA 02265470 1999-03-05wo 98/10291 PCTIGB97/02394-15-Table 2Endometrial proteins with increased synthesis inadenocarcinomaMatch # pI MW(kDa)I#l6 6.3 32I#53 6.0 109I#lll 6.7 91I#l2l 6.6 90I#l58 6.9 64I#l77 6.6 67I#l9l 6.3» 66I#l94 6.2 62I#337 6.2 45I#346 5.7 45I#436 5.4 33I#452 6.3 27I#542 6.5 103I#558 6.8 90I#627 6.9 78I#653 5.3 13I#788 6.2 130I#ll37 6.3 66I#l27l 6.3 73N#l5 8.3 32N#9l 8.1 55N#l48 8.2 44N#25l 6.6 111N#354 7.7 43N#4l4 9.5 48N#549 8.3 32N#55l 7.7 39?ICA 02265470 1999-03-05W0 98/10291 PCT/GB97/02394-15-TABLE 3Preferred endometrial proteins with increased synthesis inhyperplasiaMatch # pl MW(kDa)I#111 6.7 91I#158 6.9 64I#19l 6.3 66I#350 5.7 41I#405 5.5 35I#653 5.3 13I#892 6.6 101TABLE 4Preferred endometrial proteins with increased synthesis inadenocarcinomaMatch # pI MW(kDa)I#ll1 6.7 91I#158 6.9 64I#19l 6.3 66I#194 6.2 62I#337 6.2 45I#346 5.7 45I#4S2 6.3 27I#627 6.9 78I#653 5.3 13N#9l 8.1 55N#354 7.7 43N#55l 7.7 39?l0CA 02265470 1999-03-05W0 98/10291 PCT/GB97/02394-17-Out of the total number of approximately 1,700 spots,135 had a menstrual cycle—related expression. These 135had follows: 61proliferative intervalendometrium, 41 in secretory phase endometrium and 4 in latesecretory/menstrual Theobtained from the 2D—gel electrophoresis with respect to the(PI) (MW)subgroup of increasedspots maximal expression as spots inendometrium, 29 spots in phasephase endometrium. informationand the molecular weight of awhichsynthesis in proliferative phase endometriunt are given inisoelectric pointthese spots showpreferredTable 5 and their positions are indicated in Figures 5 and6 .?5CAWO 98/10291-18..TABLE 502265470 1999-03-05PCT/GB97/02394Endometrial proteins with menstrual cycle-related expressionMaximal expression in proliferative phase endometriumMatch # pI MW(kDa)I#1o3 6.9 86I#59O 5.4 34I#687 5.6 67I#96O 5.3 23I#1o35 6.8 52N#8 8.7 47N#2l 8.2 138N#26 6.5 124N#3l 7.7 119N#32 7.8 119N#64 8.1 66N#7l 7.1 59N#74 6.8 66N#124 7.9 48N#l92 7.7 31N#207 6.8 29N#265 7.2 70N#332 8.0 119N#342 6.7 62Fluorographs of gels exemplifying those upon which theidentifications givenappear in Figures 1 to 6.in Tables1 to 5 aboveare based?1015CA 02265470 1999-03-05W0 98/ 10291 PCT/GB97/02394-19-The above furthercharacterised by partial amino acid sequence analysis asproteins described may bedescribed in Ref. 2, or by the more sensitive technique ofmass spectrometric peptide mapping. By way of example, wehave identified the proteins for which previously givennames, data—base accession numbers and amino acid sequencesare given in Table 6. Mass spectroscopic characteristics oftryptic digests of further proteins are shown in Figures 7to 13 which have not matches to any known protein.proteinscan besequenced by known techniquesincluded per se within the scope of the invention.TABLE 6Match #NameIDAmino AcidSequenceI#l9lAndI#ll37SEQ ID~No.1Human heatshock 70 kDprotein 1PO8l07MAKAAAIGIDNDQGNRTTPSLNPQNTVFDAPFQVINDGDKSMVLTKMKEINDSQRQATKDIAYGLDRTGKTIDDGIFEVKFVEEFKRKHKKRTLSSSTQARFEELCSDLFIHDLVLVGGSKSINPDEAVALLLLDVAPLSPTKQTQIFTTKDNNLLGRFEDANGILNVTASKEEIERMVQALESYAFNMKLDKCQEVISWQVCNPIISGLSGSGPTIEEVLGTTYSCVGVYVAFTDTERLKRLIGRKFGDPKVQVSYKGEAEAYLGYPVTAGVIAGLNVLGERNVLIFDLATAGDTHLGGKDISQNKRAVSLEIDSLFEGRSTLEPVEKATRIPKVQKLLYGAAVQAAILLGLETAGGVMYSDNQPGVLILSGIPPAPRGTDKSTGKANKEAEKYKAEDESAVEDEGLKGLDANTLAEKDYQGAGGPGPGDFQHGKVEIIAIGDAAKNQVAPVVQSDMKHWTKAFYPEEISNAVITVPAYFRIINEPTAAAGGGTFDVSILEDFDNRLVNHRRLRTACERAIDFYTSITRALRDAKLDKAQQDFFNGRDLNMGDKSENVQDTALIKRNSTIQVYEGERAMTVPQIEVTFDIITITNDKGRLVQRERVSAKNKISEADKKKVEFEHKRKELEGFGAQGPKGGI#337SEQ IDNo.2CAMP-dependentproteinkinase typeI—betaregulatorychainASPPACPSEEKDCIVHLCISNRQILARQKSKARRRRGGVSYKTMTALAKAAMFPVTHIAGVDVYVNGEWVAATVKAKTDLDESLKGCELYKPERPMKFLRNSQSDSHDEEAEVYTEEDAVISKNVLFAHLETVIQQGNEGTNISEGGSFGKLWGIDRDSYVQLHGIQQVLEHFEKLEKEEVSPTPPNPVVSYVRKVIPKDDDNERSDIFDDNFYVVDQGEELALIYGTPRRRILMGSTLRand are?CA 02265470 1999-03-05WO 98/10291 PCT/GB97/02394-20_P3l32lKRKMYEEFLSEPVQFEDGEKVLQRRSPNEENRPRAATVVACSEILKRNIQKVSILESLEKIVVQGEPGDDYVEVGRLGPSRGPLKCVKLDRYNSFISLTWERLTVADRLFYIITEGTASDYFGEIALLLRPRFERVLGPI#346AndI#405SEQ IDNo.3VimentinP0867OSTRSVSSSSYTTSTRTYSLGATRSSAVRLRINTEFKNTRTRFLEQQNKILEEMRELRRQVIMRLREKLQENASLARLDLEIQELQAQIQEVRQQYESVAAANRNNDALRQALKGTNESLEIGRLQDEIQNALDIEIATYRNLRETNLDSLGQVINETSQHRRMFGGPGTASALRPSTSRSSSVPGVRLLQNEKVELQELNLAELEQLKGQDQLTNDKARVEMLQREEAENRKVESLQEEIQHVQIDVDVSKNLQEAEEWYAKQESTEYRRRQMREMEENFMKEEMARHLRKLLEGEESRIPLVDTHSKRTHDDLESRPSSSRSYVLYASSPGGVYDSVDFSLADADRFANYIDKVGKSRLGDLYEEVERDNLAEDTLQSFRQDVDAFLKKLHEEEKPDLTAALRDKSKFADLSEAQVQSLTCEVDAVEAANYQDTEYQDLLNVKMSLPLPNFSSLFLIKTVETRDI#452SEQ IDNo.4Heat Shock 27KD ProteinPO4792AndProhibitinP35232(inadmixture)MTERRVPFSLQAFGLPRLPEAAIESPAVAATADRWRVSLDTGKHEERQDETQVSSSLSPEIPVTFESRAQMAAKVFESIGDAGHRAVIFDVQKPIIFDCRTLRILFRPVASITTEILKSVSDDLTERAATEAVEAKQVAQIISAEGDSKALEAAEDIAYQLRGPSWDPFREWSQWLGGSSPAYSRALSRQVNHFAPDELTHGYISRCFTRGTLTVEAPMPLGGRSCKIRKFGLALAVAGRFRGVQDIVVSRPRNVPVITSQLPRIFTSIVARFDAGELIFGLILDDVSLQEAERARFVVAELIANSLATLSRSRNITYLDWYPHSRLFDWPGYVRPLPPLSSGVSEIRHVKTKDGVVEIKYTLPPGVDPKLATQSNEITGVVNSALYNVGEGTHFLIPWGSKDLQNVNIGEDYDERVLPTQRELVSRQVTHLTFGKEFTEKAEQQKKAAAGDGLIELRKPAGQSVLLQLPQI#436AndI#59OSEQ IDNo.5Tropomyosinfibroblastisoform TM3PO9494MDAIKKKMQMAAEDRSKQLESEALKDAQEKRIQLVEEELDDESERGMKVIAKHIAEDADRERAELSEGQVEDKYSQKEDRFAERSVTKLEHQMLDQTLLELKLDKENALDDELVSLQKKLLELAEKKATDRAQERLATALESRAQKDEEKKYEEVARKLVRQLEEQLRIMYEEEIKVLSDKSIDDLEEKVLNNMRAEQAEADKKKGTEDELDKYAEADVASLNRQKLEEAEKAAMEIQEIQLKEIIESDLERAEDQTLKALMAAKLKEAETRAEAHAKEENLSMI#627SEQ IDNo.6Serotrans—ferrinprecursorPO2787MRLAVGALLVSEHEATKCQSKASYLDCIRALAPNNLKPVVKKDSGFQMNQIGLLYCDLPEADGTDFPQLCFKCLKDGAGDQYELLCLDNTVARSMGGKEDCAVLGLCLAVFRDHMKSVIPIAANEADAVTAEFYGSKEDPLRGKKSCHTGPRKPLEKAVAQLCPGCGCSTVAFVKHSTIFRKPVDEYKDCLIWELLNQAQPDKTVRWCAVSDGPSVACVKLDAGLVYDAYQTFYYAVAVVLGRSAGWNIPNFFSGSCAPCLNQYFGYSGAENLANKADRDHLAQVPSHTVEHFGKDKSKE?CAW0 98/ 10291-21..02265470 1999-03-05PCTIGB97/02394FQLFSSPHGKKMYLGYEYVTVKWCALSHHEETTEDCIAKICGLVPVLAENVKKSASDLTWIPMGLLYNKISSLCKLCMGSRCLVEKGDVAKNLNEKDYELAPNHAVVTRKVTDCSGNFCLHDRNTYEKYLDLLFKDSAHGAIRNLREGTCRLKCDEWSVNMNGEADAMSLYNKSDNCEDTDNLKGKKSCHNHCRFDEFFSGLNLCEPNNKFVKHQTVPQNLCLDGTRKPVDKEACVHKILFRSETKDLLFGEEYVKAVGNFLKVPPRMDAPEAPTDECKPSVGKIECVSADGGFVYIAGKPEAGYFAVAVTAVGRTAGWNEGCAPGSKKDEGYYGYTGAFTGGKNPDPWAEEYANCHLARRQQQHLFGSNRDDTVCLAKLLRKCSTSSLLEACTFRRP47 KD HeatShock ProteinPrecursorN#8SEQ IDNo.7P29043MRSLLLGTLCTAEKLSSKATQAVENILVSPQAKAVLSAEQTARNVTWKLGQHYNCEHSKIDGKLPEVTKDEKFHHKMVDNLYNYYDDEKEPHHVEPLERLVAISLPKGVVKNKADLSRMSDGNPFDQDIYVRDTQSGSLLLLAVALAAEVTLAEPSTGLAVVVASSLGLVLRDEEVHAGLSRLYGPSSVSNFPDKRSALQVERTDGALLVRGFMVTRSYTKLQLVEMPLAEKLLTKEQLKEVTHDLQKHLGKKDLYLASVGREELRSPKLFIGRLVRLKGKKPVEAAAPGFSLYQAMAKDSLGGKATTASGELLRSLSNSFADDFVRSSKSINEWAAQTTNAMFFKPHWDVGVTMMHRTGHKLSSLIILMIWMGKMQKKAAGLGLTEAIDFHATAFELDTFYADHPFIFLDKMRDELN#l24 Ubiquinol-cytochrom Creductasecomplex coreprotein 2precursorSEQ IDNo.8P22695MKLLTRAGSFAPPQPQDLEFIGLFIKAGSRTKGASSFKITYTVECLRGDVVADLQPQLKIAYQNALANPLNHFTSARMALRGGLGLSGAKFVAESAVAGSRGSNTTSHLHSDSGLFGIYTIAQGNLSNTDCFLEEVGSQANADIINAAKKSRFYSLKVAPTKLPNGLVIAYEDFSNLGTTRGIEAVGGKLDILMEFLLNVDKAVAFQNPQYCPDYRIGKVIGLGVSHPVLANYRGGEIREAEANAFSVLQQAVAKATQQPISQATAAGDVVQAAKNKLKALVAGSYMPPSFVSGQKSMAAKVKATAAPAGSLENYSPVSRHLLRLTSSLTSVTATRENMATTAPEFRRWETHVIENLHAATSEELHYFVQKQVAEQFLNMQNGDSLVHAAHVLGAGPHVKFDVSAFNASYIKAAYNQVKRGYLMSVESSETVLQQIDSVASGNLGHTPFVDELN#l26 Alpha EnolasePC6733SEQ IDNo.9SILKIHAREIRAAVPSGASTVSKAVEHINKKLMIEMDGTEGAVEKGVPLYVINGGSHAGNMRIGAEVYHNGFAPNILENKIGMDVAASEFSPDQLADLYKGAWQKFTASAVNEKSCNCLLANGWGVMVSHQIKTGAPCRSKFAGRNFRNPFDSRGNPTVEGIYEALELRDTIAPALVSKKNKSKFGANAIRHIADLAGNSKLAMQEFMILLKNVIKEKYGEGLELLKTAIFRSGKYDLDFSFIKDYPVVSGIQVVGDDLTLKVNQIGSVTRSGETEDTFIERLAKYNQLLLAKVDLFTSKGLFNDKTRYMGKGLNVTEQEKIDLGVSLAVCKAEVILPVPAFNPVGAANFREAKDATNVGDEGGKAGYTDKVVKSPDDPSRYIIEDPFDQDDWVTNPKRIAKAESLQACKLAQADLVVGLCTGRIEEELGSKA?CA 02265470 1999-03-05WO 98/10291 PCT/GB97I02394_22_N#l48 Phospho— SLSNKLTLDK LDVKGKRVVMSEQ IDNo.10qlyceratekinase 1POO558RVDFNVPMKNNQITNNQRIK AAVPSIKFCLDNGAKSVVLMSHLGRPDGVP MPDKYSLEPV AVELKSLLGKDVLFLKDCVGLRFHVEEEGKASLSKLGDVYPQKAGGFLMKLGGAKVADKIFTFLKVLNNMKAEKNGVKITTVASGIPAGWKQIVWNGPVGATSRGCITIIVSTGGGASLEPEVEKACANPGKDASGNKVKVNDAFGTAHRKELNYFAKALQLINNMLDKVEIGTSLFDEELPVDFVTADKMGLDCGPESSVFEWEAFARGGGGDTATCCALLEGKVLPGVAAGSVILLENAEPAKIEAFRAHSSMVGVNLESPERPFLAINEMIIGGGMAGAKIVKDLMSFDENAKTGQAKKYAEAVTRATKALMDEVVKKWNTEDKVSHDALSNILN#207SEQ IDNo.11Triose-phosphatisomeraseISHUTS29743MAPSRKFFVGAAKVPADTEVAVAAQNCYKVWVVLGHSERRGLGVIACIGEIADNVKDWSKQAQEVHEKLRGSVTGATCKEFVDIINAKQGNWKMNGRKQVCAPPTAYIDTNGAFTGEISHVFGESDELIKLDEREAGITVVLAYEPVWAGWLKSNVSDALASQPDVDGFSLGELIGTLNFARQKLDPKIPGMIKDCGATGQKVAHALAEEKVVFEQTKVIGTGKTATPQVAQSTRIIYGLVGGASLKPEN#332SEQ IDNo.12Hypo—theticalProteinKIAAOO83P5153OPVPLSFLSTVFLGGGGLPAENRYLVLAVNTENKELCILRNSDTWIIDKDFRCMRRAVLSEVFQKAINNSFEMYRLNLSQDMHKNTSTDFPVVKPMDIEESHRGYKTKYKIVLYTLLSQERPANHLDKRELRQKTQLASLPYSRAVEQQMDQTHLEYFSLWWLMPASEMEKQYLHNFQCKHRSLFALSRGYPESTLFRLDQTFVSKKLRDLAKDTVACILKLIVGMPGTGKTSYTHSAVDNVHPAIQQFTESQLIVATTCMSQISQPICLGVLNREARALGVQYRMNSKIMNAVINLRHFKVFEPNNPVCFTEAKLIVFLTQQLKIINDLLSIVLVSFVRSTRAKHKLILLCDPRVQDGAALFQKKVVASFVQNKEGNCEKDWCSVPVEPGGYLILYPDMLTFRSSDPATRAPEKLQELAFEIKQEVEDYLQMQLSLPSDNIWSPRFGLKGMPLELKTGKERADPEAGLLLLKLRNQMAFSQIIEEEKTCKCSSVPIVMLPCLMLTLESQSSGSCIGNLIRGAIPVTNLMAVKEINMTTVTEEKNCDIDTPIIDFREPQFIGLNKPQRQAMTTTICTLVRIILLKLAKFKIQEICRSKSIKGINHPIFSRKPLFFSRRFVLMSESLFKRLESLSNKLTYEGDVKLELEFYALNTDKVPAPESIFVKAGCSPARSIGMVEVNNKDGTVGELLGCVPSLNCYPERTGAADGEEPRTVLSTGMDRLVITASQSLDIIHLEGDCTISGTSIASSIQMLIGTVLHEQTIQEIRHLKPSFCKWAGDFSKDNSTCNIEKIDVTVGVKISNSIEHRSQVYLKTGQMYPVLFHRISKSATYCSQIGNCALKIEEETQHLKKDNKKNHQNIMEHVKIVCDGGDRVIVSGEECLLDRNLSVLLGNLSKLMENSYLSSVLPHDKKVLLSKDYTLYACGFSVLLGFLRLGQIQKSLALLEELYNIFDFCIVDEAVGDHQQLPPLQNKSAVVQLTKLECGSDKVADYSDNPWLMGQVEKGGVSNVSDIGIIAPYRTVDKYQGRDKKDWRRLNVAIPLEKLLNHLN?W0 98ll0291CA-23..02265470 1999-03-05PCT/GB97l02394SEKLIIDLPSREHESLCHILGDFQREN#342SEQ IDNo.13CatalasePO404OMADSRDPASDAGNPVGDKLNMAHFDRERIPITKYSKAKVFGSADTVRDPRPIFFIRDPILMVWDFWSLRPHMNGYGSHTFGIKNLSVEDATGKYPSWTFYKVWPHKDYPLQIAFDPSNMPTHRHRLGPNYPMCMQDNQG3HSIQYSGEVRLNEEQRKRLCNFTEVHPDYGTFVQSGSHLAQMQHWKEQRAVITVGPRGPLERVVHAKGAGEHIGKKTPIAGFAVKFYTEDFPSFIHSQKRESLHQVSFLFKLVNANGEAVARLSQEDPDYIQVMTFNQAEIPVGKLVLNRPGIEASPDKMLHIPVNCPYRAPNYYPNSFGRFNTANDDNVENIAGHLKDASHIQALLDKYAREKANLAQKADVLTTGLVQDVVFTDEAFGYFEVTHDVRFSTVAGESGNWDLVGNNTNPQTHLKDPDSDRGIPDGHRYCKFHYKTDQGIRDLFNAIATFPFNPFDLTNPVNYFAEVELQGRLFAYPDARVANYQRDGAPEQQPSALETQVRAFYVNVQIFIQKKAVKNAEKPKNAIHN#55lSEQ IDNo.14Hetero-geneousnuclearribonucleo—proteinsA2/B1P22626MEKTLETVPLETTEESLRNYSRGFGFVTFSVEPKRAVAREEDTEEHHLRDKKRGFGFVTFNAEVRKALSRDSRGGGGNFGGDGYNGYGGGGGGPGYGNQGNDFGNYNQQPYGGGNYGPGGERKKREKEQFYEQWGKLTDCSMAEVDAAMAESGKPGAHVTYFEEYGKIDTDDHDPVDKIVQEMQEVQSSRPGPGSNFRGGPGGGNFGGSPGGYGGGYDNYSNYGPMKSGNSGGSGGYGGRRKLFIGGLSFVVMRDPASKRARPHSIDGRVVKKLFVGGIKIEIITDRQSGLQKYHTINGHSGRGGNFGFGSDGYGSGRGFGYGGGRGGYGGGGNYGSGNYFGGSRNMGGPSRYI#l77I#96O(Proliferativephasemarker)SEQ IDNo.15SteroidmembranebindingproteinX99714MAAEDVAATGLLYKIVRGDQDGVQDPRILMSRGLATFCLDFKYHHVGKLLADPSELEGGGPAASDSDDDEAINGKVFDVTKEALKDEYDDLLHEIFTSPLPPPLPRLKRRKGRKFYGPEGLSDLTPAQQENLLLLGLCIFDFTPAELRRFPYGVFAGRDATLNDWDSQFTKEGEEPTVYS DEEEPKDESA RKND(Hyperplasia &CancerMarker)SEQ IDNo.16shock71 KDHeatcognateproteinPl1l42MSKGPAVGIDYVAFTDTERLAVVQSDMKHWSMVLTKMKEIAGTIAGLNVLGGGTFDVSILFIAEFKRKHKSIEIDSLYEGLRDAKLDKSQQDFFNGKELNLLLLDVTPLSYSDNQPGVLIVPQIEVTFDISKEDIERMVQATVEDEKLQGEFEHQQKELEPPSGGASSGPLGT¥§§cvGvIGDAAKNQVAPFMVVNDAGRAEAYLGKTVTRIINEPTAAATIEDGIFEVKKDISENKRAVIDFYTSITRAFQHGKVEIIAMNPTNTVFDAPKVQVEYKGENAVVTVPAYFIAYGLDKKVGSTAGDTHLGGRRLRTACERARFEELNADLFIHDIVLVGGS TRIPKIQKLLKSINPDEAVALGIETAGGVMQVYEGERAMTDANGILNVSAEAEKYKAEDEKINDEDKQKIKVCNPIITKLTIEEVDYGAAVQAAILTVLIKRNTTIKDNNLLGKFEVDKSTGKENKKQRDKVSSKNLDKCNEIINWYQSAGGMPGGNDQGNRTTPSKRLIGRRFDDTKSFYPEEVSNDSQRQATKDAERNVLIFDLEDFDNRMVNHKRTLSSSTQARGTLDPVEKASGDKSENVQDPTKQTQTFTTLTGIPPAPRGITITNDKGRLSLESYAFNMKLDKNQTAEKEMPGGFPGGGAIDW(e.g.SwissProt,(PIR)Accession Identification in protein or nucleotide databasesProtein Identification Resource or EMBL)?1015CA 02265470 1999-03-05W0 98/10291 PCT/GB97/02394-24-The proteins of interest may’ be isolated from endo-metrial tissue or other protein sources by 2D gel electro-phoresis or by’ using" chromatographic techniques. Poly— ormonoclonal antibodies towards the protein of interest can beraised, and immunoassays can be established based on suchantibodies. Synthetic peptides being fragmentscharacteristic of such proteins may be used for the samepurposes. Assays may be based on more than one such proteinfor measurement at one time.Ref.1 Byrjalsen et al. Hum Reprod 1995;lO:13—18.Ref.2 Byrjalsen et al., Hum Reprod 1995;10:2760—2766.Ref.3 Julkunen et al., Endocrinology 1986;118:1782-1786.Ref.4 Byrjalsen et al., Obstet Gynecol 1992;79:523-528.Ref.5 Byrjalsen et al., Hum Reprod l992;7:lO42-1047. ?101520253035404550556065W0 98/ 10291CA 02265470 1999-03-05PCT/GB97/02394-25_SEQUENCE LISTING(1) GENERAL INFORMATION:(1) APPLICANT:(A) NAME: Center for Clinical and Basic Research(B) STREET: Ballerup Byvej 222,(C) CITY: Ballerup(E) COUNTRY: Denmark(F) POSTAL CODE (ZIP): DK—2750(ii) TITLE OF INVENTION: Biochemical Markers for the HumanEndometrium(iii) NUMBER OF SEQUENCES: 16(iv)(vi)(vi)(2)(i)MetGlnCOMPUTER READABLE FORM:(A) MEDIUM TYPE: Floppy disk(B) COMPUTER: IBM PC compatible(C) OPERATING SYSTEM: PC-DOS/MS-DOS(D) SOFTWARE: Patentln Release #l.O, Version #l.3O (EPO)A) APPLICATION NUMBER: GB 9618600.2PRIOR APPLICATION DATA:(( O6-SEP-1996B) FILING DATE:PRIOR APPLICATION DATA:(A) APPLICATION NUMBER: GB 9707l32.8(B) FILING DATE: O8-APR-1997INFORMATION FOR SEQ ID NO: 1:SEQUENCE CHARACTERISTICS:(A) LENGTH: 641 amino acids(B) TYPE: amino acid(C) STRANDEDNESS: single(D) TOPOLOGY: linearMOLECULE TYPE: proteinHYPOTHETICAL: NOANTI-SENSE: NOORIGINAL SOURCE:(A) ORGANISM: homo sapiensSEQUENCE DESCRIPTION: SEQ ID NO: 1:Ala Lys Ala Ala Ala Ile Gly Ile Asp Leu Gly Thr Thr Tyr Ser5 10 15Val Gly Val Phe Gln His Gly Lys Val Glu Ile Ile Ala Asn Asp20 25 30Gly Asn Arg Thr Thr Pro Ser Tyr Val Ala Phe Thr Asp Thr Glu35 40 _ 45?101520253035404550556065W0 98/10291AspAlaGluThr145AlaThrArgIleAsp225PheLysThrGluLeu305LeuValPheValGlyPhelle130ValGlyAlaAsnLeu210Thr‘VialArgLeuGly290CysArgGlyPheAla370ValAspTyr115AlaProValAlaVal195ThrGluAlaSer275IleSerAspGlyAsn355TyrGlyPheGlnLys100ProGluAlaIleAla180LeuIleLeuGluVal260SerAspAspAlaSer340GlyGlyCAAspAspSer85ProGluAlaTyrAla165IleAspGlyPhe245ArgSerPheLeuLys325“Pm:ArgAla02265470 1999-03-05AlaAla70AspLysGluTyrPhe150GlyAlaPheAspArgThrTyrPhe310LeuArgAspAlaAlaE.1LysMetValIleLeu135AsnLeuTyrAspGly215GluArgLeuGlnThr295ArgAspIleLeuVal375-26-LysArgLysGlnSer120GlyAspAsnGlyLeu200IleAspLysArgAla280SerSerLysProAsn360GlnAsnLeuHisVal105SerTyrSerValLeu185GlyPhePheHisThr265SerIleThrAlaLys345LysAlaGlnIleTrp90SerMetProGlnLeu170AspGlyGluAspLys250AlaLeuThrLeuGln330ValSerAlaValGly75ProTyrValValArgGlyValAsn235LysCysGluArgGlu315IleGlnIleIleAla60PheLeuThr140GlnIleThrThrLys220ArgAspGluIleAla300ProHisLysAsnLeu380GlnGlyThr125AsnAlaIleGlyPhe205AlaLeuIleArgAsp285ArgValAspLeuPCT/GB97/02394ValGlu110LysAlaThrAsnLys190AspThrValSerAla270SerPheGluLeuLeu350AspGlyIle95ThrMetValLysGlu175GlyValAlaAsnGln255LysLeuGluLysVal335GlnGluAspGlnAsp80AsnLysLysIleAsp160ProGluSerGlyHis240AsnArgPheGluAla320LeuAspAlaLys?101520253035404550556065W0 98/1029](2)Ser Glu Asn Val Gln Asp Leu Leu Leu Leu Asp Val Ala Pro Leu Ser385LeuAsnAspMetPro465AspLysGluAspTyr545LysValGluGlyGln625AspINFORMATION FOR SEQ ID NO: 2:(i)(ii)(iii)GlySerAsnThr450ProAlaAlaGluGlu530AlaIleIleHisLeu610GlyCA390Leu Glu Thr Ala405Thr Ile420Pro ThrGln Pro Gly Val435Lys Asp Asn AsnAla Pro Arg Gly470Ile485Asn Gly LeuAsn Lys Ile Thr500Ile515Glu Arg MetVal Gln Arg GluPhe Asn Met Lys550Glu Ala565Ser AspSer Trp Leu580AspLys Glu595Arg LysTyr Gln Gly AlaPro Lys Gly GlyGlyLysLeuLeu455ValAsnIleValArg535SerLysAlaLeuGly615Ser-27-GlyGlnIle440LeuProValThrGln520ValAlaLysAsnGlu600GlyGlySEQUENCE CHARACTERISTICS:(A)(B)(C)(D)LENGTH:TYPE: amino acSTRANDEDNESS:TOPOLOGY: lineidsinglearMOLECULE TYPE: proteinHYPOTHETICAL: NOVa;Thr425%GlnGlyGlnThrAsn505GluSerValLysThr585GlnProSer380 amino acidsMet410GlnValArgIleAla490AspAlaAlaGluVal570LeuValGlyGly02265470 1999-03-05395ThrIleTyrPheGlu475ThrLysGluLysAsp555LeuAlaCysProPro635AlaPheGluGlu460ValAspGlyLysAsn540GluAspGluAsnGly620ThrLeuThrGly445LeuThrLysArgTyr525AlaGlyLysLysPro605GlyIlePCT/GB97l02394IleThr430GluSerPheSerLeu510LysLeuLeuCysAsp590IlePheGluLys415TyrArgGlyAspThr495SerAlaGluLysGln575GluIleGlyGlu400ArgSerAlaIleIle480GlyLysGluSerGlyGluPheSerAlaVal640?1015202530354045SO556065W0 98/10291(iv)(vi)(xi)AlaCysCysLeuLeu65ValGlyValLysArg145GluAspIleProIle225LysSerValAsp Asp Phe Tyr Ile Ile Thr Glu Gly Thr Ala Ser Val Leu Gln ArgANTI-SENSE:CANOORIGINAL SOURCE:ORGANISM:(A)-28-homo sapiensSEQUENCE DESCRIPTION: SEQ ID NO:Ser Pro Pro Ala Cys Pro Ser GluGluIleArg50AlaSerGlyArgAla130SerThrGlnSerArg210AspArgLeuGln290LeuValGluArgProValLys115IleAspValGlyGlu195AlaArgLysGluPhe275Tyr20HisHisGlnThrSer100ValSerIleIleGlu180GlyAlaAspMetLys260Glu5ValLeuPheLysPro85AlaIleLysPheGln165ValGlyThrSerTyr245TrpAspGlnCysGluSer70ProGluPICAsnAsp150GlnAspSerValTyr230GluGluGlyLeuIleLys55AsnAsnValLysVal135AlaGlyValPheLys215ArgGluArgGlu295HisSer40LeuSerProTyrAsp120LeuMetAsnTyrGlyAlaArgPheLeuLys280GlyLysGluGlnValThr105TyrPhePheGluVal185GluLysIleLeuThr265Ile02265470 1999-03-05Glu10IleProLysSerVal90GluLysAlaProLeuThrLeuSer250ValValAspGlnGluGluAsp75LysGluThrHisVal155AspGlyAlaAspMet235LysAlaValGluGlnArgGlu60SerAlaAspMetLeu140ThrAsnGluLeuLeu220lyValAspGln300SerValPro45AsnHisArgAlaThr125AspHisPheTrpIle205LysSerSerArgGly285LeuLeu30MetArgAspArgVal110AlaAspIleTyrVal190TyrLeuThrIleLeu270GluPCT/GB97/02394Lys15LysLysGlnGluArg95SerLeuAsnAlaVal175ThrGlyTrpLeuLeu255GluP170GlyAspPheIleGluArgTyrAlaGluAsnThrGlyArg240GluProGly?WO 98/102911015(2)20253035404550556065Arg305AspThrArgIleINFORMATION FOR SEQ ID NO: 3:(i)(xi)SerProSerArgSer65AspASHPheIleSerTyrValPheGln370ProPhe'ValGlu355ArgAla340ArgTyrCAGluGlu325ArgValAsnGlu310GlyLeuSerTyrAlaProPhe375-29-Val GluLeu LeuLeu LysPro360IleSEQUENCE CHARACTERISTICS:(A)(B)(C)(D)LENGTH:TYPE:STRANDEDNESS:TOPOLOGY:465 amino acidsamino acidsinglelinearMOLECULE TYPE: proteinHYPOTHETICAL: NOANTI-SENSE: NOORIGINAL SOURCE:(A)SEQUENCE DESCRIPTION: SEQ ID NO:ORGANISM: homo sapiens345CysSerThr Arg Ser Val Ser Ser Ser SerGlyThrSer50AlaSerThrAlaLeu130ThrArg35LeuValAsn115LeuAla20ThrTyrArgAspThr100TyrAla5SerTyrAlaLeuPhe85AsnIleGluArgSerSerArq70SerGluAspLeuPITOLeuSer55SerLeuLysLysGlu135SerGly40ProSerAlaValVal120GlnSer25SerGlyValAspGlu105ArgLeuVal GlyLeu330CysSerLeu3:Tyr10SerAlaGlyProAla90LeuPheLys02265470 1999-03-05315AsnValGluThrArgArgLeuValGly75IleGlnLeuArgArqLysIleVal380ArgSerArgTyr60ValAsnGluGluGln140Leu GlyPro ArgLeu AspLeu365MetTyrPro45AlaArgThrLeuGln125Gly350LysPheVal30SerThrLeuGluAsn110GlnLysPCT/GB97/02394Pro Ser320Ala Ala335Arg ProArg AsnGly15Thr ThrThr SerSerArgLeu GlnPhe95LysAsp ArgAsn LysSer Arg?101520253035404550556065W0 98ll029lLeu145AspLeuLeuValSer225IleValGlnTrpAsn305GlnGluGluGlnLeu385LysPheValArgGlu465AlaGlnAsp210LeuGlnAspGlnTyr290AspValSerAlaAsn370LeuLeuSerAspAsp450AspLeuGluArg195AsnGlnGluValTyr275LysAlaGlnLeuAla355MetAsnLeuSerThr435GlyLeuThrAsp180GluAlaGluLeuSer260GluSerLeuSerGlu340AsnLysValGluLeu420HisGlnTyrAsn165IleGluSerGluGln245LysSerLysArgLeu325ArgTyrGluLysSerValCAGlu150Asp 7MetAlaLeuIle230AlaProValPheGln310ThrGlnGlnGluMet390GluLeuLysIleAla215AlaGlnAspAlaAla295AlaCysMetAspMet375AlaGluArgArgAsn455-30_GluAlaLeuAsn200ArgPheIleLeuAla280AspLysGluArgThr360AlaLeuSerGluThr440GluMetAJCCIArg185ThrLeuLeuGlnThr265LysLeuGlnValGlu345IleArgAspArgThr425PheThrArgVal170GluLeuAspLysGlu250AlaAsnSerGluAsp330MetGlyHisIleIle410AsnLeuSer02265470 1999-03-05Glu155GluLysGlnLeuLys235GlnAlaLeuGluSer315AlaGluArgLeuGlu395SerLeuIleGlnLeuValLeuSerGlu220LeuHisLeuGlnAla300ThrLeuGluLeuArg380IleLeuAspLysHis460ArgGluGlnPhe205ArgHisValArgGlu285AlaGluLysAsnGln365GluAlaProSerThr445HisPCT/GB97/02394Glu190ArgLysGluGlnAsp270AlaAsnTyrGlyPhe350AspTyrThrLeuLeu430Va].AspGlnValGluIle255ValGluArgArgThr335AlaGluGlnTyrPro415ProGluAspVal160AsnMetAspGluGlu24 0AspArgGluAsnArg320AsnValIleAspArg400AsnLeuThrLeu?CA 02265470 1999-03-05W0 98/ 10291 PCT/GB97/02394-31-(2)INFORMATION FOR SEQ ID NO: 4:(i) SEQUENCE CHARACTERISTICS:(A) LENGTH: 471 amino acids(B) TYPE: amino acid) STRANDEDNESS: single101520253035404550556065(ii)(iii)(iv)(vi)PheSerSerLeuValThrAspPro145GlyAsnGlyGly(C(D)TOPOLOGY:linearMOLECULE TYPE: proteinHYPOTHETICAL: NOANTI-SENSE: NOORIGINAL SOURCE:(A)SEQUENCE DESCRIPTION: SEQ ID NO:ORGANISM: homo sapiensThr Glu Arg Arg Val Pro5ProGlySer50ProArgLvsPheLeu35TrpAlaSerLeuAsp115HisGlyLeuIleSer195PheArg20ProProValGlyAsp100GlyGlyValThrThr180CysGlyAspArgGlyAlaVal85ValValTyrAspVal165IleLysLeuTrpLeuTyrAla70SerAsnValIlePro150GluProIleAlaTyrProVal55ProGluHisGluSer135ThrAlaValArgLeu215PheProGlu40ArgAlaIlePheIle120ArgGlnProThrMet200AlaSerHis25GluProTyrArgAla105ThrCysValMetPhe185AlaVal4:Leu10SerTrpLeuSerHis90ProGlyPheSerPro170GluAlaAlaLeuArgSerProArg75ThrAspLysThrSer155LysSerLysGlyArgLeuGlnPro60AlaAlaGluHisArg140SerLeuArgValGlyPheTrp45AlaLeuAspLeuGlu125LysLeuAlaAlaPhe205ValProAsp30LeuAlaSerArgThr110GluTyrSerThrGln190GluValSer15GlnGlyIleArgTrp95ValArgThrProGln175LeuSerAsnTrpAlaGlyGluGln80ArgLysGlnLeuGlu160SerGlyIleSer?CA 02265470 1999-03-05W0 98/10291 PCT/GB97/02394_32_Leu Tyr Asn Val Ala Gly His Arg Ala Val Ile Phe Arg101520253035404550ArgIleIle305IleGluLeuLeuLys385LysSerGlyGlnVal465(2)(i)Ar:ProAsnThr290PheThrThr370GlnAlaLeu450LeuTrpVal275LeuThrThrIleGlu355HisValGluAlaIle435SerLeuValSerGluThr340ArgLeuAlaGlnAla420GluArgGlnGln245GlnValIleIleIle325GlnAlaThrGlnGln405GluLeuSerLeuLysIleLeuArgAlaPheGln390LysLeuArgArgPro470IleProThrPhe295GluLysGluThrLysIleLysAsn455GlnINFORMATION FOR SEQ ID NO: 5:ValIleAspSerLeuPhe360LysAlaAlaAlaLeu440IleSEQUENCE CHARACTERISTICS:ValIle265SerProTyrValVal345GlyGluGluAlaAsn425GluThrGly250PheLysValAspVal330SerLeuPheArgIle410SerAlaTyr235GluAspAspAlaGlu3l5AlaArgIleThrAla395IleLeuAlaLeuGlyCysLeuSer300ArgArgGlnLeuGlu380ArgSerAlaGluPro460Th:ArgGln285GlnValPheValAsp365AlaPheAlaThrAsp445AlaHisSer270AsnLeuLeuAspSer350AspValValGluAla430IleGlyProProAla335AspValGluValAlaGln240LeuProAsnArgSer320GlyAspSerAlaGlu400AspAspTyrSer(A) LENGTH: 284 amino acids(B) TYPE: amino acid(C) STRANDEDNESS: single(D) TOPOLOGY: linear(ii) MOLECULE TYPE: protein(iii).HYPOTHETICAL: NOANTI-SENSE: NOORIGINAL SOURCE:(A) ORGANISM: homo sapiens?WO 98/10291-33-(xi) SEQUENCE DESCRIPTION: SEQ ID NO:Met Asp Ala Ile Lys Lys Lys Met Gln5 1 5Asn Ala Leu Asp Arg Ala Glu Gln Ala20 251O Glu Asp Arg Ser Lys Gln Leu Glu Asp35 40Lys Leu Lys Gly Thr Glu Asp Glu Leu50 5515Lys Asp Ala Gln Glu Lys Leu Glu Leu65 70Ala Glu Ala Asp Val Ala Ser Leu Asn20 85Glu Glu Leu Asp Arg Ala Gln Glu Arg100 10525Leu Glu Glu Ala Glu Lys Ala Ala Asp115 12030 Val Ile Glu Ser Arg Ala Gln Lys Asp130 135Glu Ile Gln Leu Lys Glu Ala Lys His145 1503 5Lys Tyr Glu Glu Val Ala Arg Lys Leu165Glu Arg Ala Glu Glu Arg Ala Glu Leu40 180 185Leu Glu Glu Gln Leu Arg Ile Met Asp195 20045 Ala Ala Glu Asp Lys Tyr Ser Gln Lys210 215Ile Lys Val Leu Ser Asp Lys Leu Lys225 23050Phe Ala Glu Arg Ser Val Thr Lys Leu245Glu Glu Lys Val Ala His Ala Lys Glu55 260 265Met Leu Asp Gln Thr Leu Leu Glu Leu275 2806065CA02265470 1999-03-055:Met10GluGluAspAlaArg90LeuGluGluIleVal170SerGln: GluGluGlu250GluAsnLeuAlaLeuLysGlu75ArgAlaSerGluAla155IleGluThrAspAla235LysAsnAsnLysAspValIleThrGluLys140GluIleGlyLeuArg220GluSerLeuMetLeuLysSer45SerLysGlnAlaArg125MetAspGluGlnLys205TyrThrIleSerPCTIGB97/02394AspLys30LeuGluAlaLeuLeu110GlyGluAlaSerVal190AlaGluArgAspMet270Lys15AlaGlnAlaThrVal95GlnMetIleAspAsp175ArgLeuGluAlaAsp255HisGluAlaLysLeuAsp80GluLysLysGlnArg160LeuGlnMetGluGlu240LeuGln?CA 02265470 1999-03-05W0 98/10291 PCT/GB97/02394-34-INFORMATION FOR SEQ ID NO: 6:SEQUENCE CHARACTERISTICS:(A) LENGTH: 698 amino acids) TYPE: amino acidSTRANDEDNESS:(1)singlel01520253035404550556065HisIleLeu65LeuLysPheAsnAla145ProCysCysGlyLys22(B(C)(D)TOPOLOGY:linearMOLECULE TYPE: proteinHYPOTHETICAL: NOANTI-SENSE: NOORIGINAL SOURCE:ORGANISM: homo sapiens(A)SEQUENCE DESCRIPTION: SEQ ID NO:ArgLeuGluPro50AspTyrGln130GlyArgAlaProAla210HisLeu Ala Val Gly Ala Leu LeuAlaAla35SerCysAlaValTyr115LeuTrpLysPICPheSerVal20ThrAspIleGlyVal100AlaArgAsnProCys180CysLysThr5ProLysGlyArgLeu85AlaValGlyIleLeu165AlaGlyCysIleAspCysProAla70ValGluLysPro150GluAspCysLeuPhe230LysGlnSer55IleTyrPheValLys135IleLysGlySerLys215GluThrSer40ValAlaAspTyrVal120SerGlyAlaThrThr200AspAsnVal25PheAlaAlaAlaCysLeuValAsp185LeuGlyLeuVal10ArgArgCysAsnTyr90SerLysHisLeuAla170PheAsnAlaAlaCysTrpAspValGlu75LeuLysAspThrTyr155AsnProGlnGlyAsn235AlaCysHisLys60AlaAlaGluSerGly140CysPheGlnTyrAsp220LysValAlaMet45LysAspProAspAspPheLeuPhe205ValAlaLeuVal30LysAlaAlaAsnPro110PheGlyLeuSerCys190GlyAlaAspGly15SerSerSerValAsn95GlnGlnArgProTyrPheArgLeuGluValTyrThr80LeuThrMetSerGlu160SerLeuSerValAsp240?CA 02265470 1999-03-05W0 98/10291 PCT/GB97/02394-35-Gln Tyr Glu Leu Leu Cys Leu Asp Asn Thr Arg Lys Pro Val Asp Glu245 250 2555 Tyr Lys Asp Cys His Leu Ala Gln Val Pro Ser His Thr Val Val Ala260 265 270Arg Ser Met Gly Gly Lys Glu Asp Leu Ile Trp Glu Leu Leu Asn Gln275 280 28510Ala Gln Glu His Phe Gly Lys Asp Lys Ser Lys Glu Phe Gln Leu Phe290 295 300Ser Ser Pro His Gly Lys Asp Leu Leu Phe Lys Asp Ser Ala His Gly15 305 310 315 320Phe Leu Lys Val Pro Pro Arg Met Asp Ala Lys Met Tyr Leu Gly Tyr325 330 33520 Glu Tyr Val Thr Ala Ile Arg Asn Leu Arg Glu Gly Thr Cys Pro Glu340 345 350Ala Pro Thr Asp Glu Cys Lys Pro Val Lys Trp Cys Ala Leu Ser His355 360 36525His Glu Arg Leu Lys Cys Asp Glu Trp Ser Val Asn Ser Val Gly Lys370 375 380Ile Glu Cys Val Ser Ala Glu Thr Thr Glu Asp Cys Ile Ala Lys Ile30 385 390 395 400Met Asn Gly Glu Ala Asp Ala Met Ser Leu Asp Gly Gly Phe Val Tyr405 410 41535Ile Ala Gly Lys Cys Gly Leu Val Pro Val Leu Ala Glu Asn Tyr Asn420 425 430Lys Ser Asp Asn Cys Glu Asp Thr Pro Glu Ala Gly Tyr Phe Ala Val40 435 440 445Ala Val Val Lys Lys Ser Ala Ser Asp Leu Thr Trp Asp Asn Leu Lys450 455 46045 Gly Lys Lys Ser Cys His Thr Ala Val Gly Arg Thr Ala Gly Trp Asn465 470 475 480Ile Pro Met Gly Leu Leu Tyr Asn Lys Ile Asn His Cys Arg Phe Asp485 490 49550Glu Phe Phe Ser Glu Gly Cys Ala Pro Gly Ser Lys Lys Asp Ser Ser500 505 510Leu Cys Lys Leu Cys Met Gly Ser Gly Leu Asn Leu Cys Glu Pro Asn55 515 520 525Asn Lys Glu Gly Tyr Tyr Gly Tyr Thr Gly Ala Phe Arg Cys Leu Val530 535 54060Glu Lys Gly Asp Val Ala Phe Val Lys His Gln Thr Val Pro Gln Asn545 550 555 560Thr Gly Gly Lys Asn Pro Asp Pro Trp Ala Lys Asn Leu Asn Glu Lys65 565 570 575Asp Tyr Glu Leu Leu Cys Leu Asp Gly Thr Arg Lys Pro Val Glu Glu580 585 590?10152025WO 98/10291(2)TyrArgHis625PheLeuGluLeuINFORMATION FOR SEQ ID NO: 7:(1)LeuArgAlaTyrLeu690Asn595AspPheSerLysVal675GluCysLysGlyGluLeu660LysAlaHisGluSerThr645HisAlaCysCALeuAlaAsn630LysAspValThrAlaCys615ValAspArgGlyPhe695-35-Arg600ValThrLeuAsnAsn680ArgSEQUENCE CHARACTERISTICS:AlaHisAspLeuThr665LeuArgProLysCysPhe650TyrArgPro02265470 1999-03-05AsnIleSer635ArgGluLysHisLeu620GlyAspLysCysAla605ArgAsnAspTyrSer685PCT/GB97/02394ValGlnPheThrLeu670ThrValGlnVal655SerThrGlnLeu640CysGluSerLENGTH: 417 amino acidsTYPE: amino acidSTRANDEDNESS: single(A)30354O45SO556065(xi)MetAlaGluLeuAsn65SerSerLeu(B)(C)(D)TOPOLOGY:linearMOLECULE TYPE: proteinHYPOTHETICAL: NOANTI-SENSE: NOORIGINAL SOURCE:ORGANISM: homo sapiens(A)SEQUENCE DESCRIPTION:SEQ ID NO:Arg Ser Leu Leu Leu Gly Thr LeuAlaLysAla50IleLeuAlaLeuGluLeu35PheLeuGlyGluArg115Val20SerSerValGlyGln100Ser5LysSerLeuSerLys85LeuLeuLysLysTyrPro70AlaArgSerProAlaGln55ValThrAspAsnValThr40AlaValThrGluSer120Glu25ThrMetValAlaGlu105Thr7:Cys10AlaLeuAlaAlaSer90ValAlaLeuAlaAlaLysSer75GlnHisArgLeuAlaGluAspSerAlaAlaASHAlaProPro45GlnLeuLysGlyVal125ValGly30SerAlaGlyAlaLeu110ThrAla15ThrThrValLeuVal95GlyTrpLeuAlaGlyGluVal80LeuGluLys?CA 02265470 1999-03-05W0 98/ 10291 PCT/GB97/02394_37-Leu Gly Ser Arg Leu Tyr Gly Pro Ser Ser Val Se: Phe Ala Asp Asp130 135 140 45 Phe Val Arq Ser Ser Lys Gln His Tyr Asn Cys Glu His Ser Lys Ile145 150 155 160Asn Phe Pro Asp Lys Arg Ser Ala Leu Gln Ser Ile Asn Glu Trp Ala165 170 17510Ala Gln Thr Thr Asp Gly Lys Leu Pro Glu Val Thr Lys Asp Val Glu180 185 190Arg Thr Asp Gly Ala Leu Leu Val Asn Ala Met Phe Phe Lys Pro His15 195 200 205Trp Asp Glu Lys Phe His His Lys Met Val Asp Asn Arg Gly Phe Met210 215 22020 Val Thr Arg Ser Tyr Thr Val Gly Val Thr Met Met His Arg Thr Gly225 230 235 240Leu Tyr Asn Tyr Tyr Asp Asp Glu Lys Glu Lys Leu Gln Leu Val Glu245 250 25525Met Pro Leu Ala His Lys Leu Ser Ser Leu Ile Ile Leu Met Pro His260 255 270His Val Glu Pro Leu Glu Arg Leu Glu Lys Leu Leu Thr Lys Glu Gln30 275 280 285Leu Lys Ile Trp Met Gly Lys Met Gln Lys Lys Ala Val Ala Ile Ser290 295 30035 Leu Pro Lys Gly Val Val Glu Val Thr His Asp Leu Gln Lys His Leu305 310 315 320Ala Gly Leu Gly Leu Thr Glu Ala Ile Asp Lys Asn Lys Ala Asp Leu40 325 330 335Ser Arg Met Ser Gly Lys Lys Asp Leu Tyr Leu Ala Ser Val Phe His340 345 350Ala Thr Ala Phe Glu Leu Asp Thr Asp Gly Asn Pro Phe Asp Gln Asp45 355 360 365Ile Tyr Gly Arg Glu Glu Leu Arg Ser Pro Lys Leu Phe Tyr Ala Asp370 375 38050 His Pro Phe Ile Phe Leu Val Arg Asp Thr Gln Ser Gly Ser Leu Leu385 390 395 400Phe Ile Gly Arg Leu Val Arg Leu Lys Gly Asp Lys Met Arg Asp Glu405 410 41555Leu60 (2) INFORMATION FOR SEQ ID NO: 8:(i) SEQUENCE CHARACTERISTICS:(A) LENGTH: 453 amino acids(B) TYPE: amino acid65 (C) STRANDEDNESS: single(D) TOPOLOGY: linear(ii) MOLECULE TYPE: protein?101520253035404550556065CA 02265470 1999-03-05WO 98/10291(xi)MetLysProIleIle65HisPheThrAspGlu145AspLeuProValVal225ArgGluAla-38-HYPOTHETICAL: NOANTI-SENSE: NOORIGINAL SOURCE:(A) ORGANISM: homo sapiensSEQUENCE DESCRIPTION: SEQ ID NO: 8:Lys Leu Leu Thr Arg Ala Gly Ser Phe5 10Ala Thr25Val Ala Pro Lys Val Lys Ala20Phe Thr40Gln Pro Gln Leu Glu35Asp LysAla Glu Asn Ser Pro Val50Ser Leu Tyr55Lys Ala Gly Ser Arg Glu Asp Phe70TyrThr90Leu Thr Ser Ser Leu85Leu Leu ArgGlu Ala105Thr Ile Val100Lys Ile Arg GlyAla120Ala Thr Glu Asn Met Thr115Arg TyrGlu Phe Leu Leu135Val130Asp Ile Leu MetGlu Ala Leu150Arg Arg Trp Val AspGln170Ala Val Ala Phe Gln Asn Pro165LysAla185Ala Ala Ala Gln Asn Leu180His TyrVal Thr200Asp Tyr Ile Gly Ser195Arg LysPhe Thr Ser Ala Met215Gln210Asn His ArgVal Leu Gln Val Ala230Ser His Pro LysGly Ala Lys250Gly Leu Leu SerSer Leu265Glu Asn260Arg Gly AspSer Ala Glu280Ser Ala Ala275GlySerAlaLeuSerSer75ThrGlyValASHGln155ThrAlaGluAlaGlu235AlaValAlaArgProProArg60AsnLysGlyGluVal140ProHisAsnGluLeu220GlnAsnHisAsnPCT/GB97/02394Phe Tyr SerAlaAsn45IleLeuGlyLysCys125ThrGlnValProLeu205IlePheTyrAlaAla285GlyGlyGlyGlyAlaLeu110LeuThrLeuIleLeu190HisGlyLeuArgAla270Phe15AlaLeuLeuThrSer95SerArgAlaLysGlu175TyrTyrLeuAsnLeuProValPheThr80SerValGlyProIle160AsnCysPheGlyMet240GlyValVal?101520253035404550556065CA 02265470 1999-03-05W0 98/1029](2)LeuThr305PheGlyAlaThrSer385LeuAspSerPhe-39-Val Ala295Gln His Leu290Gly Fro HisHis Gln Val Ala310Thr Ser His LeuAla325Val Ser Phe Asn Ala Ser Tyr330AspThr345Thr Ala Ala340Ile Tyr Ile Ser GlnAla Tyr Asn Gln Val Ile Ala355Lys Arg360Val Gln Ala Ala Lys Asn Leu375Asp370Phe GluGlu Glu390Val Ser Ser CysSer410Val Ala Gly Ser Met Pro405TyrVal Ala Asn Ala Ile Asn420Ser AspAla440Gln435Gly Lys Ser Met Ala GlyVal450Asp Glu LeuINFORMATION FOR SEQ ID NO: 9:(i)(ii)(iii)(iv)(vi)ProAlaArgSEQUENCE CHARACTERISTICS:(A) LENGTH: 433 amino acids(8) TYPE: amino acid(C) STRANDEDNESS: single(D) TOPOLOGY: linearMOLECULE TYPE: proteinHYPOTHETICAL: NOANTI-SENSE: NOORIGINAL SOURCE:(A) ORGANISM: homo sapiensSEQUENCE DESCRIPTION: SEQ ID NO: 9:Ile Leu Lys Ile His Ala Arg Glu Ile5 10Thr Val Glu Val Asp Leu Phe Thr Ser20 25Val Pro Ser Gly Ala Ser Thr Gly Ile35 40Asp Asn Asp Lys Thr Arg Tyr Met Gly50 55PCT/GB97/02394Val Lys300Lys Ala Thr Gln315Ser Asp Ser GlyAla Val350Gly AspGln Asn Leu365GlyAla380Lys Gly TyrGlu Val395Gly SerThr Val Leu GlnAla Ala Lys Lys430HisAsn LeuPhe Asp Ser ArgLys Gly Leu Phe30Glu Ala45Tyr LeuLys Gly Val Ser60GlnLeu335IleSerLeuGlnGln415PheThrGly Asn15Arg AlaGlu LeuLys AlaArg Gly Ser AsnLysAsnMetAla400IleValPro?1015202530354045SO556065W0 98/ 10291LeuAspValLeuPro145LysPheAsnGluLeu225IleAspAspValLys305ValAsnLeuSerVal385GluGluAsnGlySerTyr130ValLeuArgValGly210LeuLeuGlnSer290PheThrCysGlnHis370GlyArgValThrLeu115ArgProAlaGluIle195GlyLysMetAspLeu275IleThrASHLeuAla355ArgLeuLeuIleThrGlu100AlaHisAlaMetAla180LysPheThrAspPhe260AlaGluAlaProLeu340CysSerCysAlaAsnGlu85AsnValIlePheGln165MetGluAlaAlaVal245LysAspAspSerLys325LeuLysGlyThrLys405CALys70GlnLysCysAlaAsn150GluArgLysProIle230AlaSerLeuPIOAla310ArgLysLeuGluGly390TyrThrSerLysAsp135ValPheIleTyrAsn215GlyAlaProTyrPhe295GlyIleValAlaThr375GlnAsn-40-IleLysLysAla120LeuIleMetGlyGlyIleLysSerAspLys280AspIleAlaAsnGln360GluIleGlnAlaIlePhe105GlyAlaAsnIleAla185LysLeuAlaGluAsp265SerGlnGlnLysGln345AlaAspLysLeuAspGlyAlaGlyGlyLeu170GluAspGluGlyPhe250ProPheAspValAla330IleAST1Leu41002265470 1999-03-05Ala75AlaValAsnGly155ProValAlaAsnTyr235PheSerIleAspVal315ValGlyGlyPhe-Gly395ArgLeuLeuAsnGluSer140SerValTyrThrLys220ThrArgArgLysTrp300GlyAsnSerTrpIle380AlaIleValMetAlaLys125GluHisGlyHisAsn205GluAspSerTyrAspGluValGly365AlaProGluSerIle110GlyValAlaAlaAsn190ValIle270TyrAlaAspLysThr350ValAspCysGluGlu95LeuValIleGlyAla175LeuGlyLeuValLys255SerProTrpLeuSer335GluMetLeuArgGlu415PCT/GB97/02394ProLeuAsn160AsnLysAspGluVal240TyrProValGlnThr320CysSerValValSer400Leu?W0 98/1029]1520253035404550556065CA-41-02265470 1999-03-05PCT/GB97l02394Gly Ser Lys Ala Lys Phe Ala Gly Arg Asn Phe Arg Asn Pro Leu AlaLysINFORMATION FOR SEQ ID NO:(i)(ii)(iii)(xi)SerArgIleCysArg65AlaAspGlyGlyLys145ValGly..................——.__.....s,.,,42010:SEQUENCE CHARACTERISTICS:(A)(B)(C)(D)LENGTH:TYPE:STRANDEDNESS:TOPOLOGY:417 amino acidsamino acidsinglelinearMOLECULE TYPE: proteinHYPOTHETICAL: NOANTI-SENSE: NOORIGINAL SOURCE:homo sapiens(A)SEQUENCE DESCRIPTION:ORGANISM:425SEQ ID NO:10:Leu Ser Asn Lys Leu Thr Leu Asp LysValThrLeu50ProValCysSerLys130IleAsnValValAsn35AspAspGluValVal115GlyGluAspAsnMet20AsnAsnGlyLeuGly100IleLysAlaAlaLeu1805ArgGlnGlyValLys85ProLeuAspPhePhe165ProValArgAlaPro70SerGluLeuAlaArg150GlyGlnAspIleLys55MetLeuValGluSer135AlaThrLysPheLys40SerLeuGluAsn120GlySerAlaAlaAsn25AlaValAspGlyLys105LeuAsnLeuHisGly18510ValAlaValLysLys90AlaArgLysSerArg170GlyLeuProValLeuTyr75AspCysPheValLys155AlaPheAspMetProMet60SerValAlaHisLys140LeuHisLeuValLysSer45SerLeuLeuAsnVal125AlaGlySerMet430LysAsn30IleHisGluPhePro110GluGluAspSerLys190Gly15AsnLysLeuProLeu95AlaGluProValMet175LysGlnPheVal80LysAlaGluAlaTyr160ValGlu?CA 02265470 1999-03-05PCT/GB97/02394l015'20253035404550556065W0 98/10291(2)Leu.1‘*4L1:Asn225PhePheGluAspGlyAlaIleGlu385LeuLeuINFORMATION FOR SEQ ID NO:(i)ThrAspLysLys290IleLysProLeuIle370AspLeuTyr195LeuLeuPheGluAsn275PheProTyrValMet355GlyLysGluAspLeuGlu260GlyAspAlaAlaGly340AspGlyValGlyLysLys245GlyValGluGlyGlu325ValGluGlySerLys405LysAlaVal230ValAlaLysAsnTrp310AlaPheValAspHis390ValAlaLys215AsnLeuLysAla295MetValGluValThr375ValLeu-42-Leu200ValGluAsnIleThr280LysGlyThrTrpLys360AlaSerProll:SEQUENCE CHARACTERISTICS:(A)(B)(C)(D)LENGTH:TYPE:amino acidSTRANDEDNESS:TOPOLOGY:linesinglarMOLECULE TYPE: proteinHYPOTHETICAL: NOANTI-SENSE: NOORIGINAL SOURCE:ORGANISM:(A)SEQUENCE DESCRIPTION:Ala Pro Ser Arg Lys Phe Phe Val Gly Gly Asn Trp Lys Met Asn5249 amino acidsehomo sapiensGluAlaMetASHVal265LeuThrLeuArgGlu345AlaThrThrGlySEQ ID NO:SerAspIleMet250LysProGlyAspAla330AlaThrCysGlyVal410ll:10ProLysIle235GluAspValGlnCys315LysPheSerCysIleLeuAspAla300GlyGlnAlaArgAla380GlyAlaArg205GlyGlyMetPhe285ThrProIleArgGly365LysAlaLeuLe;GlyThr276ValValGluValTrpSerSerPheIleMetSer255LysThrAlaSerTrp335ThrIleAsnLeuAsn41515LeuAsnAla240LeuAlaAlaSerSer320AsnLysThrThrGlu400Ile?CA 02265470 1999-03-05wo 93/10291 PCT/GB97/02394-43-Gly Arq Lys Gln Ser Leu Gly Glu Leu Ile Gly Th: Leu Asn Ala Ala20 25 305 Lvs Val Pro Ala Asp Thr Glu Val Val Cys Ala Pro Pro Thr Ala Tyr35 40 45Ile Asp Phe Ala Arg Gln Lys Leu Asp Pro Lys Ile Ala Val Ala Ala50 55 6010Gln Asn Cys Tyr Lys Val Thr Asn Gly Ala Phe Thr Gly Glu Ile Ser65 70 75 soPro Gly Met Ile Lys Asp Cys Gly Ala Thr Trp Val Val Leu Gly His15 85 90 95Ser Glu Arg Arg His Val Phe Gly Glu Ser Asp Glu Leu Ile Gly Gln100 105 11020 Lys Val Ala His Ala Leu Ala Glu Gly Leu Gly Val Ile Ala Cys Ile115 120 125Gly Glu Lys Leu Asp Glu Arg Glu Ala Gly Ile Thr Glu Lys Val Val130 135 14025Phe Glu Gln Thr Lys Val Ile Ala Asp Asn Val Lys Asp Trp Ser Lys145 150 155 160Val Val Leu Ala Tyr Glu Pro Val Trp Ala Ile Gly Thr Gly Lys Thr30 165 170 175Ala Thr Pro Gln Gln Ala Gln Glu Val His Glu Lys Leu Arg Gly Trp180 185 19035 Leu Lys Ser Asn Val Ser Asp Ala Val Ala Gln Ser Thr Arg Ile Ile195 200 205Tyr Gly Gly Ser Val Thr Gly Ala Thr Cys Lys Glu Leu Ala Ser Gln210 215 22040Pro Asp Val Asp Gly Phe Leu Val Gly Gly Ala Ser Leu Lys Pro Glu225 230 235 24045 Phe Val Asp Ile Ile Asn Ala Lys Gln245(2) INFORMATION FOR SEQ ID NO: 12:50 (i) SEQUENCE CHARACTERISTICS:(A) LENGTH: 1076 amino acids(B) TYPE: amino acid(C) STRANDEDNESS: single(D) TOPOLOGY: linear55(ii) MOLECULE TYPE: protein(iii) HYPOTHETICAL: NO60 (iv) ANTI-SENSE: NO(vi) ORIGINAL SOURCE:(A) ORGANISM: homo sapiens65.,»..._....-...-—.-...-............4.....«...........m.~ ..... , .?CA 02265470 1999-03-05PCT/GB97l023941015‘202530354045SO556065W0 98/10291GlySerVal65ArgIleIleAspSer145ThrValGluLeuGlu225MetProLysLysLys305SEQUENCE DESCRI PTION:ValGlyGlyPhe50LeuLeuLeuHisPhe130IlePheLeuLysLys210ValHisSerProGly290Thr-44-SEQ ID NO:Pro Leu Ser Phe Leu Ser ThrAlaGly35ProAlaValArgLeu115GlyAlaArgHisLeu195GluGluMet275Ala20GlyArgValIleAsn100GluTyrSerSerGlu180GlnMetAspAsnAsn260AspIleTyrL‘)GluLeuThrAsnThr85AspGlyLeuSerSer165ValGluTyrTyrThr245SerIleAspLysArgProValThr70AlaTrpAspIleIle150AspPheLeuArgLeu230SerLysGluValIle310ThrAlaLeu55ValSerCysCysLeu135ArgProGlnAlaLeu215PIOThrAspGluThr295MetGlyGlu40SerGlnGlnSerThr120TyrCysAlaLysPhe200AsnSerAspAsnSer280ValProAla25LeuThrAsnSerVal105SerProMetThrAla185GlnLeuPhePheSer265IleGlyLeuVal10AlaPheGlyLysLeu90ProAspAspArgArg170IleThrSerCysPro250ThrTrpValGlu12:CysAspGlnMetGlu75GluValThrMetArg155GlnAsnIleGlnLys235GlnCysSerLysLeu315AspGlyLysAsp60GlyAsnGluTrpLeu140AlaMetAsnGlnAspTrpMetAsnProIle300LysProGluLys45AsnAsnLysProIle125IleValLeuSerGlu205GluAlaGlnIleArg285HisThrValArgCysGluSerLeuIlePhe190IleIleGlyLeuGlu270PheArgGly#4 <25-.PheValTyrGluLeu95AspAspGlySerArgLysAspSer255ValGlyGlyLysGlnLeuAlaLeuLys80CysIleLysThrGlu160ThrProHisGlnPhe240LeuValLeuTyrGlu320?WO 98/10291Ser Asn5 Ser GL1Lys ThrlOGlu Leu370Ile Ser15 385Gln Ile20 Asn CysSer val2 5Leu Lys450Thr Leu30 465Trp Leu35 Asn LeuLeu His4 0Met Ala530Ala Leu45 545Cys Leu50Arg LeuAsn Leu5 5Asp Leu6106 0Ser Val625Gly Leu6 5Lys AspLysIleAlaPro435GlnGluMetIleAsn515GlySerLeuAspSer595IleLeuAsnTyrIleArg340GlnLysSerGluLeu420IleThrSerProArg500PheAspArgAspGln580LysIleProLysThr660CAGlu325ArgMetLeuAlaGlu405TyrValHisGlnAla485MetGlnArgGlyArg565GluLeuAspHisPro645Leu02265470 1999-03-05HisAlaTyrArgThr390GluSerMetLeuSer470SerGluCysValTyr550ASHGluMetPheAsp630GlnIleArgAspProAsn375ArgLysArgLeuGlu455LysGluHisLysIle535ValLeuLysGluArg615AlaArgVal-45-SerProVal360GlnGlnThrAlaPro440TyrAspMetValHis520ValLysSerASHAsn600GluLysGlnGlyGlnGlu345ProMetLysCysVal425LysPheAsnGluLys505GlySerGluProAspAlaMet665Val330Ala«AlaAlaThrLys410GluIleSerLysLys490IleAlaGlyIleLeu570AspPheGlnThrMet650Pro(X)H<AsnPheGln395TyrGlnGluLeuLys475SerValIleGluASH555ProIleValPheVal635LysGlyLeuSer380LeuCysGlnGluTrp460AsnGlyCysGlu540MetAspSerIle620AlaLysThrTyrLeuLeu365LeuAlaSerMetGlu445CysHisSerAspVal525ArgThrSerThrLys605SerCysValGlyPCT/GB97/02394ThrL4) F’Ln (I:O LIPheSerGlnAsp430ThrLeuGlnCysSerThrThr5§oLysTyrIleLeuLys670Leu335TyrLysHisLeuIle415CysGlnMetAsnIle495GlnAsnLeuValLeu575LeuLeuLeuLeuLeu655ThrLeuLeuArgArgPro400GlySerHisLeuIle480GlyTyrLeuPheThr560PheGlyArgSerLys640SerThr?101520253035404550556065W0 98/10291Leu705ValLysLeuArgGln785ValArgLysIleTrpThrAsnLys945GlnValValAlaHisSerIleLys770ProGlyAlaSerMet850SerValVal930AlaGlnGluLeuCys675ThrLysProIleVal755IleIleAspLeuAla835SerAspLysMetLys915ThrGlyLeuValVal995ThrSerPheAlaLys740AlaPheCysHisGly820ValLeuLysLeuGluCysLysAsn980SerLysIle725SerThrAspLeuGln805MetValSerValGlu885ValProAlaSerIle965ThrPheCAValTh:Ile710GlnLeuThrPheSerGlnAsnAla870LeuPheAlaLysPro950IleValValArgHis695GlyGlnAlaCysCys775ProLeuGluLeuLys855AsnGluGluProLeu935SerAsnAspArg-46Ile680SerPhePheLeuMet760IleLeuProSerThr840LeuAlaPheProGlu920IleAspAspLysSer1000LeuAlaLeuThrLeu745GlyValPheProLeu825ValThrValTyrAsn905GlnValIleLeuTyr985AsnTyrValArgGlu730GluIleAspPheLeu810PheGlnTyrIleAla890AsnValPheGlyLeu970GlnLys02265470 1999-03-05AlaAspLeu715GlnGluAsnGluSer795ValLysTyrGluAsn875AspProGluLeuIle955AlaGlyAspAsn700GlyGluLeuHisAla780ArgLeuArgArgGly860LeuTyrValLysThr940IleArgArgGlyIleTyrPro765SerMet845LysArgSerCysSerAlaSerAspThr1005PCT/GB97/02394PheCysAsn750IleGlnPheArgGlu830AsnLeuHisAspPhe910GlyIleProIleLys990ValArg735SerPheIleValGlu815GlnSerGluPheAsn895LeuValPheTyrValLysLys720SerGlnSerSerLeu800AlaAsnLysCysLys880ProAsnSerValArg960MetIleGlu?101520253035404550556065CA 02265470 1999-03-05W0 98/ 10291-47-Leu Leu Lys Asp Trp Arg Arg Leu Asn Val Ala Ile Thr10201010 1015His1025 1030 1035Pro Leu Glu Lys Leu Leu Asn His Leu Asn Ser Glu Lys1045 1050Asp Leu Pro Ser Arg Glu His Glu Ser Leu Cys His Ile1060 1065Phe Gln Arg Glu1075(2)INFORMATION FOR SEQ ID NO: 13:(i) SEQUENCE CHARACTERISTICS:(A) LENGTH: 527 amino acids(B) TYPE: amino acid(C) STRANDEDNESS: single(D) TOPOLOGY: linearMOLECULE TYPE: proteinHYPOTHETICAL: NOANTI-SENSE: NOORIGINAL SOURCE:(A) ORGANISM: homo sapiens(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 13:Met Ala Asp Ser Arg Asp Pro Ala Ser Asp Gln Met1 5 10Glu Gln Ala20Arg Ala Gln Lys Ala Val Leu ThrVal Thr35Asn Ile Val40Asn Pro Gly Asp Lys LeuVal Gln Val55Val Phe Thr Glu60Leu Leu50Pro Asp AspHis Ala75Glu Ile Pro Glu Val Val70Asp Arg Arg ArgAla Phe Gly Phe Glu Val Thr His Ile Thr85Tyr Asp90Ala Val Phe Thr100Lys Glu His Ile Gly Lys Pro105LysPhe Thr Ala115Val Ala Gly Glu Ser Ser120Gly AspPro Arg Phe Ala Val Phe Thr Glu130Gly Lys135Tyr Asp140Val Thr150Ile Phe Phe Ile155Leu Asn Asn Pro145Gly ArgLys Leu Ile Leu Leu Gly Cys Val Pro Ser Leu AsnGlnThrGly45MetLysLysIleThr125GlyAspPCT/GB97/02394Arg Ala LysLeu Ile Ile1055Leu Gly Asp1070HisGlyProAlaGlyTyrAla110ValAsnProTrp15AlaArgHisAlaSer95ValArgTrpIleLysGlyGlyPheGlyLysArgAspAspLeu160?1015.20253035404550556O65WO 98/10291PheLysLeuHisAla225GlyAspLysAlaHis305AsnSerGlyAsnAsn385AlaSerAsnAspHisArg210AsnIleProTyrGlu290LysProAsnArgTyr370ProAlaThrVal450PheAspGln195HisGlyLysAspPro275ThrAspValMetLeu355LeuGlnAsnLeuAla435LeuLeuThrLysPheAsp180ValMetGluAsnTyr260SerPheTyrAsnPro340PheHisArgTyrGlu420AsnASHLysGluTyr500CAIle165MetSerAsnAlaLeu245GlyTrpProProTyr325ProAlaIleAspTyr405HisAspGluAspVal485AsnHisValPheGlyVal230SerIleThrPheLeu310PheGlyTyrProSerAspGluAla470SerTrpLeuTyr215TyrValArgPheAsn295IleAlaIleProVal375ProAsnIleAsnGln455GlnProGlu-48-GlnAspPhe200GlyCysGluAspTyr280ProProGluGluAsp360AsnMetSerGlnVal440ArgIleAspLysLysPhe185SerSerLysAspLeu265IlePheValValAla345ThrCysCysPheTyr425ThrLysPheTyrPro505HisPheAla250PheGlnAspGlyGlu330SerHisProMetGly410SerGlnGly490Lys02265470 1999-03-05ThrHis235AlaAsnValLeuLys315GlnProArgTyrGln395AlaGlyValLeuGln475SerAsnLeu .GlyPhe220TyrArgAlaMetThr300LeuIleAspHisArg380AspProGluArgCys460LysHisAlaIle205LysLysLeuIleThr285LysValAlaLysArg365AlaAsnGluValAla445GluLysIleIleLeuThrSerAla270PheValLeuPheMet350LeuArgGlnGlnArg430PheAsnAlaGlnHis510PCTIGB97/02394AspValAspGln255ThrAsnTrpASHAsp335LeuGlyValGlyGln415ArgTyrIleValAla495ThrLeuSerGlyASHGln240GluGlyGlnProArg320ProGlnProAlaPheValAlaLys480LeuPhe?W0 98/ 1029110152O253035404550556065Val Gln Ser Gly Ser His Leu Ala Ala Arg Glu Lys Ala Asn LeuINFORMATION FOR SEQ ID NO:(1)(xi)Met,4LysThrAspGly65AlaValLysArgThr145AspH,_.(DMetPhe515CA02265470 1999-03-05-49-52014:SEQUENCE CHARACTERISTICS:353 amino acids(A)(B)(C)(D)LENGTH:TYPE:STRANDEDNESS:TOPOLOGY:amino acidsinglelinearMOLECULE TYPE: proteinHYPOTHETICAL: NOANTI-SENSE: NOORIGINAL SOURCE:ORGANISM: homo sapiens(A)SEQUENCE DESCRIPTION: SEQ ID NO:GluGluGluCys50PheArgAlaLeuAsp130AspAspAsnGlnGlyLysGlnGlu35ValValProArgPhe115TyrArgHisGly1Glu195AspThrPhe20SerValThrHisGlu100ValPheGlnAspHis180ValSerLeu5ArgLeuMetPheSer85GluGlyGluSerPro165AsnGlnArgGluLysArgArgSer70IleSerGlyGluValAlaSerGlyThrLeuAsnAsp55SerAspGlyIleTyr135LysAspGluSerValPheTyr40ProMetGlyLysLys120GlyLysArg200GlyProIle25TyrAlaAlaArgPro105GluLysArgIleArg185SerGly14:Leu10GlyGluSerGluVal90GlyAspIleGlyVal170LysGlyAsnGluGlyGlnLysVal75ValAlaThrAspPheI55LeuAlaArgPheArgLeuTrpArg60AspGluHisGluThr140GlyGlnLeuGly525LysSerGly45SerAlaProValGlu125IlePheLysSerGlyProPCT/GB97/02394AlaLysThr110HisGluValTyrArg190AsnGlyArg15GluLeuGlyMetArg95ValHisIleThrHis175GlnPheProGluThrThrPheAlaAlaLysLeuIlePhe160ThrGluGlyGly?101520253035404550556065W0 98/ 10291(2)Ser225GlyGlyAsnAsn305PheGlyTyrINFORMATION FOR SEQ ID NO:(i)(vi)(Xi)MetGluLeuAspAsnAspGlyProTyr290TyrGlyProPheGlySerAsnGlyGlyArgTyrPro260TyrGlyGlnSerGly340GlyAsn245GlyGlyGlnArg325SerCAGly230Gly ”TyrAsnAsnPro310AsnGlySerGlyGlnTyr295SerMetGly-50-02265470 1999-03-05Asp Gly Tyr GlyASHGlySer15:SEQUENCE CHARACTERISTICS:A)B)C)D(((( )LENGTH:TYPE:STRANDEDNESS: singleTOPOLOGY:194 amino acidsamino acidlinearMOLECULE TYPE: proteinHYPOTHETICAL: NOANTI-SENSE: NOORIGINAL SOURCE:ORGANISM: homo sapiens(A)SEQUENCE DESCRIPTION: SEQ ID NO:AlaAlaGlyLeu35ProLeuValGluGly20GlyAlaLysGlnAsp Val Ala5LeuLeuAlaArg ’Asp85LeuCysSerHisIleAspAspArgAlaGluPheSerPheIleGlySerTyrGlyThrIle25LeuAspThrLeuGlyGlyGlyPro330Gly15:GlyPheLeuAspProMetProGlyTyrAsnPro315TyrTyrAlaThrTyrAspAla75AlaSerGlyGlyGlyTyr300MetGlyGlyAspSerLysGlu60GluIleGly285AsnLysGlyGlyProProIle45ProLeuAsnPCT/GB97/02394Gly2GlyAspSerGlyArg350SerLeu30ValProArgGlyAsn255GlyTyrPheGlyAsn335SerGlu15AsnArgProArgLys95Phe240PheGlyAspAsn320TyrArgLeuLeuGlyLeuPhe80Val?CA 02265470 1999-03-05W0 93/10291 PCT/GB97/02394-51-Phe Val Thr Glu100Asp Gly Lys Phe Pro105Arg Tyr Gly Pro Ty:Ala125Va; Phe Leu115Ala Ser120Gly la Gly Arg Asp Arg Gly CysGlu Ala Leu Glu Tyr Asp Leu Leu140Leu Lys Lys135Asp Asp Asp Asp130Thr Gln Thr Ser G1n Phe145Ala Gln Glu150Leu Asn Tn;160Pro Asp Trp155AspVal Glu Glu Glu175Leu170His Leu Gly165Phe Lys Tyr His Gly Lys Lys proAla190Thr Glu Glu Glu Pro Glu Ser185Ser180Val Tyr Asp Lys Asp Arg LysASDAsn(2) INFORMATION FOR SEQ ID NO: 16:SEQUENCE CHARACTERISTICS:(A) LENGTH: 646 amino acidsTYPE: amino acidsinglelinear(i)TOPOLOGY:3035404550556065GlnArgAsnAlaAspSer()(C) STRANDEDNESS:(D)MOLECULE TYPE: proteinHYPOTHETICAL: NOANTI-SENSE:ORIGINAL SOURCE:(A)SEQUENCE DESCRIPTION: SEQ ID NO:NOORGANISM: homo sapiensSer Lys Gly Pro Ala Val Gly IleValGlyLeu50ThrValAlaPheGlyAsn35IleValValGlyTyr115Val20ArgGlyPheGlnArg100PIC5PheThrAspAspSer85ProGlnThrAlaAla70AspLysGluHisProAla55LysMetValValGlySer40LysArgLysGlnSer120AsnLeuHisVal105Ser16:Asp10ValValGlnIleTrp90GluMetLeuGluAlaValGly75ProTyrValGlyIlePheAla60ArgPheLysLeuThrIleThrMetArgMetGlyThr125ThrAlaAspAsnPheValGlu110LysTyr15AsnThrProAspVal95ThrMetSerGluThrAsp80AsnLysLys?CA 02265470 1999-03-05wo 98/10291 PCT/GB97/02394-52-Glu Ile Ala Glu Ala Tyr Leu Gly Lys Thr Val Thr H Val Val1O1520253035404550556O65Thr145AlaThrArgIleAsp225PheLysThrGluLeu305LeuValPheValSer385LeuAsnAspMetPro465130ValAlaAsnLeu210ThrIleArgLeuArgGlyPheAla370GluGlyThrAsnThr450ProProThrAlaVal195ThrHisAlaAlaSer275IleAlaAspGlyAsn355TyrAsnIleThrGln435LysAlaAlaIleAla180LeuIleLeuGluVal260SerAspAspAlaSer340GlyGlyValGluIle420ProAspProTyrAla165IleIleGluPhe245ArgSerPheLeuLys325ThrLysAlaGlnThr405ProGlyAsnArgPhe150GlyAlaPheAspGlyLysArgThrTyrPhe310LeuArgGluAlaAsp390AlaThrValAsn135AsnLeuTyrAspArgLeuGlnThr295ArgAspIleLeuVal375LeuGlyLysLeuLeu455ValAspAsnGlyLeu200IleAspLysArgAla280SerGlyLysProAsn360GlnLeuGlyGlnIle440LeuProLeu185GlyPhePheHisThr265IleThrSerValThr425GlnGlyGlnGlnLeu170AspGlyGluAspLys250AlaIleThrLeuGln330IleSerAlaLeuMet410GlnValLysIleArg155ArgLysGlyValAsn235LysCysGluArgAsp315IleGlnIleIleAsp395ThrThrTyrPheGlu475140G‘"IleLysThrLys220ArgAspGluIleAla300ProHisLysAsnLeu380ValValPheGluGlu460ValIleValPhe205SerMetIleArgAsp285ArgValAspLeuPro365SerThrLeuThrThrASHThrValSerAla270SerPheGluIleLeu350AspGlyProIleThr430GluThrPhesysGlu175AlaValAlaAsnGlu255LysLeuGluLysVal335GlnGluAspLeuLys415TyrArgGlyAspAsp160ProGluSerGlyHis240AsnArgTyrGluAla320LeuAspAlaLysSer400ArgSerAlaIleIle480?l015202530W0 98/10291AspGluAspTyr545LysIleGluLysPhe625ThrAlaGluAspGlu530AlaHisLeu610AsnAsnIle515LysPheAsnAsnGln595TyrGlyGluGlyLys500GluGlnAsnAspTrp580GlnGlnGlyGluCAIle485IleArgArgMetGlu565LeuLysSerGlyVal645LeuThrMetAspLys550AspAspGluAlaAla630AspAsnIleValLys535AlaLysLysLeu-53-ValThrGln520ValThrGlnAsnGlu600GlyProSearAsn505GluSeerValLysGln585LysMetSer02265470 1999-03-05Ala490AspAlaSerGluIle570ThrValProValLysGluLysAsp555LeuAlaCysGlyGlyAspGlvLysAsn540GluAspGluAsnGly620AlaLysArgTyr525SerLysLysLysPro605MetSerPCT/GB97/02394SerLeu510LysLeuLeuCysGlu590IleProSe]:GluGlnAsn575GluIleGlyGlyGlyLysGluSerGly560GluPheThrGlyPro640

Claims (16)

CLAIMS:

1. A method of characterising a biological sample comprising detecting or quantitating therein one or more proteins produced by the endometrium in increased amounts in hyperplasia or in adenocarcinoma as shown by 2D gel electrophoresis comparison of cell lysates of endometrial biopsies from normal endometrium and endometrium showing hyperplasia or adenocarcinoma, excluding variations due to the menstrual cycle, or detecting or quantitating a fragment or breakdown product thereof, or a nucleic acid coding therefor or antibodies thereto.

2. A method of characterising a biological sample comprising detecting or quantitating therein one or more proteins produced by the endometrium in increased amounts in hyperplasia or in adenocarcinoma and characterised by one of the following combinations of molecular weight and pI
values:

hyperplasia pI MW kDa 6.7 91 6.6 90 6.9 64 6.6 67 6.3 66 6.8 46 5.7 41 5.5 35 5.3 13 6.6 101 5.8 14 7.4 8.2 44 9.5 98 adenocarcinoma pI MW (kDa) 6.3 32 6.0 109 6.7 91 6.6 90 6.9 64 6.6 67 6.3 66 6.2 62 6.2 45 5.7 45 5.4 33 6.3 27 6.5 103 6.8 90 6.9 78 5.3 13 6.2 130 6.3 66 6.3 73 8.3 32 8.1 55 8.2 44 6.6 111 7.7 43 9.5 48 8.3 32 7.7 39 or a fragment or breakdown product thereof, or a nucleic acid coding therefor or antibodies thereto.

3. A method as claimed in Claim 1 or Claim 2, wherein said protein, fragment, breakdown product, antibodies, or nucleic acid is detected in a body fluid sample.

4. An immunological binding partner specifically reactive with a protein as defined in Claim 1 or Claim 2 or with a fragment or breakdown product thereof or with a nucleic acid coding therefor.

5. A cell line producing a monoclonal antibody being an immunological binding partner as claimed in Claim 4.

6. An assay kit for use in a method as claimed in Claim 1 or Claim 2, comprising an immunological binding partner as claimed in Claim 4.

7. A method of characterising a biological sample comprising detecting or quantitating therein one or more proteins produced by the endometrium in increased amounts during the proliferative phase of the endometrium as shown in 2D gel electrophoresis comparison of cell lysates of endometrial biopsies from normal endometrium in its proliferative and secretory phases and characterised by one of the following combinations of molecular weight and pI
values:-pI MW(kDa) 6.9 86 5.4 34 5.6 67 5.3 23 6.8 52 8.7 47 8.2 138 6.5 124 7.7 119 7.8 119 8.1 66 7.1 58 6.8 66 7.9 48 7.7 31 6.8 29 7.2 70 8.0 119 6.7 62 or a fragment or breakdown product thereof, or a nucleic acid coding therefor, or an antibody thereto.

8. A method as claimed in Claim 7, for detecting the phase of the endometrium.

9. A method as claimed in Claim 7 or Claim 8, wherein said protein, fragment, or breakdown product is detected in a body fluid sample.

10. An immunological binding partner specifically reactive with a protein as defined in Claim 7 or with a fragment or breakdown product thereof or with a nucleic acid coding therefor.

11. A cell line producing a monoclonal antibody being an immunological binding partner as claimed in Claim 10.

12. An assay kit for use in a method as claimed in Claim 7 or Claim 8, comprising an immunological binding partner as claimed in Claim 10.

13. A protein produced by the endometrium in increased amounts in hyperplasia or in adenocarcinoma as shown by 2D gel electrophoresis comparison of cell lysates of endo-metrial biopsies from normal endometrium and endometrium showing hyperplasia or adenocarcinoma, excluding variations due to the menstrual cycle, and characterised by one of the following combinations of molecular weight and pI values:

hyperplasia pI MW kDa 6.7 91 6.6 90 6.9 64 6.8 46 5.7 41 5.3 13 6.6 101 5.8 14 9.5 48 adenocarcinoma pI MW (kDa) 6.3 32 6.0 109 6.7 91 6.6 90 6.9 64 6.2 62 6.5 103 6.8 90 5.3 13 6.2 130 6.3 66 6.3 73 8.3 32 8.1 55 6.6 111 7.7 43 9.5 48 8.3 32

14. A protein produced by the endometrium in increased amounts during the proliferative phase of the endometrium as shown in 2D gel electrophoresis comparison of cell lysates of endometrial biopsies from normal endometrium in its proliferative and secretory phases and characterised by one of the following combinations of molecular weight and pI values:-pI MW(kDa) 6.9 86 5.6 67 6.8 52 8.2 138 6.5 124 7.7 119 7.8 119 8.1 66 7.1 58 6.8 66 7.7 31

15. A protein as claimed in Claim 13 or Claim 14, characterised by the properties:-PI MW(kDa) 5.7 41 5.6 67 9.5 48 6.8 52 6.5 124 7.7 119 7.8 119 and by the respective tryptic digestion MS spectra shown in Figures 7 to 12.

16. The use of a protein as defined in any one of Claims 1, 2 or 7 or a fragment thereof, for detecting autoantibodies to a said protein.