多肽设计原理和方法PPT课件.ppt

.,1,多肽设计原则和方法,.,2,多肽抗原的特点,含有的epitope数目少；抗原性比融合蛋白弱；产生的抗原特异性通常比融合蛋白强；是一种半抗原，需要连接载体蛋白；,.,3,多肽设计的原则,Hydrophilicity,tobesuretheantigenisonthesurfaceoftheprotein.Antigenicity,aminoacidprovidesgoodbonds.N-orC-terminalpeptide,fortheyareexposedinmostproteins.ContainsMHCIIpreferredmotifs.SuchasD,Y,P,andFwhoappearsinMHCmoleculesmostfrequently.Nohomologywithotherproteinsbyblast.Preferthecytoplasmicdomainforatransmembraneprotein,fortheyaremoreimmunogenicAvoidglycosylationandphosphorylationsitesifpossibleAvoidcystininthemiddlepartofthepeptide,.,4,20种氨基酸的抗原性比较,.,5,什么时候选用多肽做抗原,我们没有基因的抗原；有基因，但是蛋白质和其他蛋白质有较强的同源性，或者蛋白质属于同源性较强的蛋白家族；有基因，需要区分该基因编码的蛋白质加工后的不同亚型；抗原是修饰蛋白质，包括磷酸化，乙酰基化蛋白质等,.,6,案例1我们没有的蛋白质,NM_004445，EPHB6MATEGAAQLGNRVAGMVCSLWVLLLVSSVLALEEVLLDTTGETSEIGWLTYPPGGWDEVSVLDDQRRLTRTFEACHVAGAPPGTGQDNWLQTHFVERRGAQRAHIRLHFSVRACSSLGVSGGTCRETFTLYYRQAEEPDSPDSVSSWHLKRWTKVDTIAADESFPSSSSSSSSSSSAAWAVGPHGAGQRAGLQLNVKERSFGPLTQRGFYVAFQDTGACLALVAVRLFSYTCPAVLRSFASFPETQASGAGGASLVAAVGTCVAHAEPEEDGVGGQAGGSPPRLHCNGEGKWMVAVGGCRCQPGYQPARGDKACQACPRGLYKSSAGNAPCSPCPARSHAPNPAAPVCPCLEGFYRASSDPPEAPCTGPPSAPQELWFEVQGSALMLHWRLPRELGGRGDLLFNVVCKECEGRQEPASGGGGTCHRCRDEVHFDPRQRGLTESRVLVGGLRAHVPYILEVQAVNGVSELSPDPPQAAAINVSTSHEVPSAVPVVHQVSRASNSITVSWPQPDQTNGNILDYQLRYYDQAEDESHSFTLTSETNTATVTQLSPGHIYGFQVRARTAAGHGPYGGKVYFQTLPQGELSSQLPERLSLVIGSILGALAFLLLAAITVLAVVFQRKRRGTGYTEQLQQYSSPGLGVKYYIDPSTYEDPCQAIRELAREVDPAYIKIEEVIGTGSFGEVRQGRLQPRGRREQTVAIQALWAGGAESLQMTFLGRAAVLGQFQHPNILRLEGVVTKSRPLMVLTEFMELGPLDSFLRQREGQFSSLQLVAMQRGVAAAMQYLSSFAFVHRSLSAHSVLVNSHLVCKVARLGHSPQGPSCLLRWAAPEVIAHGKHTTSSDVWSFGILMWEVMSYGERPYWDMSEQEVLNAIEQEFRLPPPPGCPPGLHLLMLDTWQKDRARRPHFDQLVAAFDKMIRKPDTLQAGGDPGERPSQALLTPVALDFPCLDSPQAWLSAIGLECYQDNFSKFGLCTFSDVAQLSLEDLPALGITLAGHQKKLLHHIQLLQQHLRQQGSVEV共1021aa，N-15aa为信号肽,.,7,案例1我们没有的蛋白质（2）,WDEVSVLDDQRRLTRTFEAC,.,8,案例1我们没有的蛋白质（3）,GENEID:2051EPHB6|EPHreceptorB6HomosapiensQuery1WDEVSVLDDQRRLTRTFEAC20WDEVSVLDDQRRLTRTFEACSbjct41WDEVSVLDDQRRLTRTFEAC60GENEID:814CAMK4|calcium/calmodulin-dependentproteinkinaseIVQuery1WDEVS-VLDDQRRLTRTFEA19WDEVSVLD+RLTTF+ASbjct69WDEVSLNAKDLVRKLIVLDPKKRLT-TFQA97,.,9,案例2家族性蛋白的多肽设计（transgelin）,Thetransgelinfamilyisagroupofproteinsthatbelongto22kdactin-relatedcorpninsuperfamily.Ofallthreeisoforms,transgelin1isthebestcharacterized.Transgelin1,alsoknownasSM22alpha,isaspecificmarkerfordifferentiatedsmoothmusclecells.Transgelin2,alsoknownasSM22beta,isexpressedbybothsmoothmuscleandnon-smoothmusclecellsinatemporallyandspatiallyregulatedpattern.Trangenlin3,alsoknownasNP25,isonlyfoundinhighlydifferentiatedneuronalcells.,.,10,案例2家族性蛋白的多肽设计（transgelin）2,.,11,案例2家族性蛋白的多肽设计（transgelin）3,Transgelin1:KYDEELEERLVEWIIVQCTransgelin2:QPGRENFQNWLKDGTVLC,.,12,案例3家族性蛋白的多肽设计RAB3A,3B,3C,Sequence2:NM_002866,RAB3A,memberRASoncogenefamilySequence3:NM_002867,RAB3B,memberRASoncogenefamilySequence4:NM_138453,RAB3C,memberRASoncogenefamily,.,13,案例4家族性蛋白的多肽设计actin,对应的处理方法？,.,14,案例5同一蛋白质不同亚型的多肽设计NFKB1,NFKB1,NUCLEARFACTORKAPPA-B,SUBUNIT1;NFKB1Alternativetitles;symbolsTRANSCRIPTIONFACTORNFKB1NUCLEARFACTOROFKAPPALIGHTCHAINGENEENHANCERINBCELLS1NFKBp105,INCLUDEDNFKBp50,INCLUDED,.,15,案例5同一蛋白质不同亚型的多肽设计NFKB1,Thisgeneencodesa105kDproteinwhichcanundergocotranslationalprocessingbythe26Sproteasometoproducea50kDprotein.The105kDproteinisaRelprotein-specifictranscriptioninhibitorandthe50kDproteinisaDNAbindingsubunitoftheNF-kappa-B(NFKB)proteincomplex.1433aminoacidsoffulllength968p105proteinformp50.,.,16,案例5同一蛋白质不同亚型的多肽设计NFKB1,MAEDDPYLGRPEQMFHLDPSLTHTIFNPEVFQPQMALPTADGPYLQILEQPKQRGFRFRYVCEGPSHGGLPGASSEKNKKSYPQVKICNYVGPAKVIVQLVTNGKNIHLHAHSLVGKHCEDGICTVTAGPKDMVVGFANLGILHVTKKKVFETLEARMTEACIRGYNPGLLVHPDLAYLQAEGGGDRQLGDREKELIRQAALQQTKEMDLSVVRLMFTAFLPDSTGSFTRRLEPVVSDAIYDSKAPNASNLKIVRMDRTAGCVTGGEEIYLLCDKVQKDDIQIRFYEEEENGGVWEGFGDFSPTDVHRQFAIVFKTPKYKDINITKPASVFVQLRRKSDLETSEPKPFLYYPEIKDKEEVQRKRQKLMPNFSDSFGGGSGAGAGGGGMFGSGGGGGGTGSTGPGYSFPHYGFPTYGGITFHPGTTKSNAGMKHGTMDTESKKDPEGCDKSDDKNTVNLFGKVIETTEQDQEPSEATVGNGEVTLTYATGTKEESAGVQDNLFLEKAMQLAKRHANALFDYAVTGDVKMLLAVQRHLTAVQDENGDSVLHLAIIHLHSQLVRDLLEVTSGLISDDIINMRNDLYQTPLHLAVITKQEDVVEDLLRAGADLSLLDRLGNSVLHLAAKEGHDKVLSILLKHKKAALLLDHPNGDGLNAIHLAMMSNSLPCLLLLVAAGADVNAQEQKSGRTALHLAVEHDNISLAGCLLLEGDAHVDSTTYDGTTPLHIAAGRGSTRLAALLKAAGADPLVENFEPLYDLDDSWENAGEDEGVVPGTTPLDMATSWQVFDILNGKPYEPEFTSDDLLAQGDMKQLAEDVKLQLYKLLEIPDPDKNWATLAQKLGLGILNNAFRLSPAPSKTLMDNYEVSGGTVRELVEALRQMGYTEAIEVIQAASSPVKTTSQAHSLPLSPASTRQQIDELRDSDSVCDSGVETSFRKLSFTESLTSGASLLTLNKMPHDYGQEGPLEGKI,.,17,案例5同一蛋白质不同亚型的多肽设计NFKB1,.,18,案例6同一蛋白质不同亚型的多肽设计CXCR3,TheCXCR3proteinbelongstoGprotein-coupledreceptorfamily.Severalalternativelysplicedtranscriptvariantsencodingdistinctisoformshavebeenreported.TheCXCR3Bproteincontains416aminoacidsandhasalongerNterminusthatdiffersfromtheoriginal368-aminoacidCXCR3protein,CXCR3A,inthefirst52residues.Thisantibody,60065-1-Ig,isspecificforCXCR3B,notbindingCXCR3A.,