人EEN蛋白的结构预测和功能分析3

156199912ChineseJournalofBiochemistryandMolecularBiologyVol.15,No.6Dec.,19993(39800026)(7992026):,,19642,,Tel:[phone],Fax:[phone]E2mail:[e-mail]:1999201229,:1999204223EEN31)1)1)2)(1),100850,2),200025)EEN(extraelevennineteen,EEN),:EEN,NAB,A,CSH3,SEM25öGRB2CSH3;,EENSH3,:EENSH3SEM25öGRB2SH3,.:EEN,,SH3.EEN,SH3,,StructurePredicationandFunctionAnalysisofHumanEENProteinLIUMengmin,ZHANGShaowen,HEFuchu(InstituteofRadiationMedicine,AcademyofMilitaryMedicalScience,Beijing100850)CHENZhu(ShanghaiInstituteofHeamotology,Shanghai200025)AbstractThesecondarystructureofthededucedaminoacidsequencesofpreviouslyclonedhu2manEENgenehasbeenpredicatedbyneuralnetworkmethod.TheresultsshowthatEENpro2teinmaycontainthreedomains:TheNH22terminalregioncontainsthreeAhelicesandBsand2wich,thecentralregioncontainsfourAhelices,andtheCOOH2terminalregioncontainsanSrchomology3(SH3)domainmostsimilartotheC2terminalSH3domainfoundintheSEM25öGRB2familyofgenesinvolvedinreceptortyrosinekinasesignaling.The32dimensionalstructureofSH3domainofEENproteinhasbeenconstructedbyhomologymodelingmethod.Theresultsshowthatthe32dimensionalstructureofEENSH3domainismostsimilartothatofSEM25öGRB2SH3domain,andtheaminoacidsequencesofSH3domainwhichbindsproline2richmotifsinligandproteinarehighlyconservedbetweenEENproteinandSEM25öGRB2proteins.Thesere2sultssuggestthatEENproteinmaybeanewsignalprotein,involvedinnewsignaltransductionpathwayorpara2pathway,andSH3domainisitsfunctionaldomain.KeywordsEEN,SH3domain,Structureprediction,Signaltransduction,.3:.(),,[1].,,,[2].,,.HRX[homologofdrosophilatrithoraxormul2tiplelineageleukemia(MLL)orALL21]11q23,,,[3].EEN(extraelevennineteen,EEN)HRX,HRX,;EEN368,,19p13[4].,EEN,.[5],EEN,[6],EENSH3.EEN.1111EENFASTA(PDB)EEN,:EEN,CSEM25[7](),GRB2[8](),53BP2[9](p532)CSH3,EENCSH3;,EENCSH3SEM25GRB2CSH3,4517%3517%,53BP2SH3(2816%),Fig.1SH3,,SH3,28,EENSH340%.Holley[5]EEN,,.Fig.1SequencealignmentoftheSH3domainbetweenEENproteinandotherrelatedproteinsTheidenticalresiduesareconnectedbysolidlines;theconservedresiduesareconnectedbydashedlines112EENSH3,Sali[6],SEM25,EENSH3.QUANTA,PROTEINDESIGN,MODELLERCHARMM.2211EEN,EEN,CSH3,SEM25GRB2CSH3,4517%3517%,SEM25GRB2SH3,,[10],EEN.Fig.2,,EEN:N(M1G101),A(M1S14,L29V42,E90G101),BLoop,AB9496:EENFig.2SecondarystructureofwholeEENproteinpredictedwithneuralnetworkprogram;(G105S248)A(G105F139,D141Q175,K177N203,D208S248),;C(P299Q368)SH3,BB.,S249T298SH3Loop(B),LoopLoop,EEN.Fig.2,(AB),.EEN,,.212EENSH3,,SEM25,EENFig.3Schematicdiagramofthe3dimensionalstructureofEENSH3domainmodelledwithMODELERprogram.ThesidechainsshowtheaminoacidsformingthebindingsiteofprolineSH3,Fig.3.,,,05915,.,,,[6].EENSH3SEM25SH34517%,,EENSH3.Fig13,,SEM25.SH3Table1(SEM25).SH3,Fig14Ramachandran,,SH357Ramachandran,SH3.Fig.4TheRamachandranplotofthestructureofSH3do2mainmodelledbyMODELERprogramTable1Importantresiduesformingtheproline2bindingsiteofSH3domainsProteinsResiduesnumber(indicatedinFig.1)192128506366EENYFEWPYGRB2FFEWPYSEM25FFEWPY53BP2WYEWPLTable1,6,28Glu,5.SH3.GRB2SEM256,EEN19PheTyr,,.EENSH3SEM25GRB2SH3,,,,.p532(53BP2),192166TrpTyrLeu,53BP2SH3SH3,.53BP2,SH3,[9].,4SH3,,,;EENCSH3SEM25öGRB2CSH3,,:,,.References1RabbittsTH.Translocations,mastergenes,anddifferencesbetweentheoriginsofacuteandchronicleukemiascancer.Cell,1991,67:6416442RabbittsTH.Chromosomaltranslocationsinhumancancer.Nature,1994,372(6502):1431493TrachukDC,KohlerS,ClearyML.InvolvementofhomologofDrosophilatrithoraxby11q23chromosomaltranslocationsinacuteleukemias.Cell,1992,71:6917004SoCW,CaldasC,LiuMM,ChenSJ,HuangQH,GuLJ,ShamMH,WiedemannLM,ChanLC.EENencodesforamemberofanewfamilyofproteinscontaininganSrchomology3domainandisthethirdgenelocatedonchromosome19p13thatfusestoMLLinhumanleukemia.ProcNatlAcadSciUSA,1997,94:256325685HolleyLH,KarplusM.Proteinsecondarystructurepredictionwithaneuralnetwork.ProcNatlAcadSciUSA,1989,86:1521566SaliA,MatsumotoR,McNeilHP,KarplusM,StevensRL.Three2dimensionalmodelsoffourmousemastcellchymases.ldentificationofproteoglycanbindingregionsandprotease2spe21596:EENcificantigenicepitopes.JBiolChem,1993,268:902390347LimWA,RichardsFM,FoxRO.Structuraldeterminantsofpeptide2bindingorientationandofsequencespecificityinSH3domains.Nature,1994,372:3753798KohdaD,TerasawaH,IchikawaS,OguraK,HatanakaH,MandiyanV,UllrichA,SchlessingerJ,InagakiF.Solutionstructureandligand2bindingsiteofthecarboxy2terminalSH3domainofGRB2.Structure,1994,2(11):102910409GorinaS,PavletichNP.StructureoftheP53tumorsuppres2sorboundtotheankyrinandSH3domainsof53BP2.Science,1996,274:1001100510PawsonT.Proteinmodulesandsignallingnetworks.Nature,1995,373:57358014,NIH,6,10L?923,14L920,IncyteRandallScott:142634LNealLaneRobertMay,()L,Incyte?Scott,IncyteCpG(CpGislands)LCpGDNA,L,IncyteCpG,L,53%CpGL75000CpG,142634LL,ScottCpGL,,,20%L,3109,L,IncyteL,(TheGuardian),L,,LWellcomeTrust();,,24L,LL,WellcomeTrustSangerJohnSulston,,L,(NIH)L,(ESTS)cDNA,cDNA,LcDNALDNA,(NCI)RobertStrausberg;LNature(401:311418,1999)25915