多肽設(shè)計原則和方法1.多肽抗原的特點含有的epitope數(shù)目少；抗原性比融合蛋白弱；產(chǎn)生的抗原特異性通常比融合蛋白強；是一種半抗原，需要連接載體蛋白；2.多肽設(shè)計的原則Hydrophilicity,tobesuretheantigenisonthesurfaceoftheprotein.Antigenicity,aminoacidprovidesgoodbonds.N-orC-terminalpeptide,fortheyareexposedinmostproteins.ContainsMHCIIpreferredmotifs.SuchasD,Y,P,andFwhoappearsinMHCmoleculesmostfrequently.Nohomologywithotherproteinsbyblast.Preferthecytoplasmicdomainforatransmembraneprotein,fortheyaremoreimmunogenicAvoidglycosylationandphosphorylationsitesifpossibleAvoidcystininthemiddlepartofthepeptide3.20種氨基酸的抗原性比較L0.75K-0.05H0.65P-0.05D0.31R-0.07T0.21W-0.11A0.12C-0.12E0.06N-0.14Y0.01G-0.18V-0.01M-0.38S-0.01F-0.77Q-0.03I-2.924.什么時候選用多肽做抗原我們沒有基因的抗原；有基因，但是蛋白質(zhì)和其他蛋白質(zhì)有較強的同源性，或者蛋白質(zhì)屬于同源性較強的蛋白家族；有基因，需要區(qū)分該基因編碼的蛋白質(zhì)加工后的不同亞型；抗原是修飾蛋白質(zhì)，包括磷酸化，乙?；鞍踪|(zhì)等5.案例1我們沒有的蛋白質(zhì)NM_004445，EPHB6MATEGAAQLGNRVAGMVCSLWVLLLVSSVLALEEVLLDTTGETSEIGWLTYPPGGWDEVSVLDDQRRLTRTFEACHVAGAPPGTGQDNWLQTHFVERRGAQRAHIRLHFSVRACSSLGVSGGTCRETFTLYYRQAEEPDSPDSVSSWHLKRWTKVDTIAADESFPSSSSSSSSSSSAAWAVGPHGAGQRAGLQLNVKERSFGPLTQRGFYVAFQDTGACLALVAVRLFSYTCPAVLRSFASFPETQASGAGGASLVAAVGTCVAHAEPEEDGVGGQAGGSPPRLHCNGEGKWMVAVGGCRCQPGYQPARGDKACQACPRGLYKSSAGNAPCSPCPARSHAPNPAAPVCPCLEGFYRASSDPPEAPCTGPPSAPQELWFEVQGSALMLHWRLPRELGGRGDLLFNVVCKECEGRQEPASGGGGTCHRCRDEVHFDPRQRGLTESRVLVGGLRAHVPYILEVQAVNGVSELSPDPPQAAAINVSTSHEVPSAVPVVHQVSRASNSITVSWPQPDQTNGNILDYQLRYYDQAEDESHSFTLTSETNTATVTQLSPGHIYGFQVRARTAAGHGPYGGKVYFQTLPQGELSSQLPERLSLVIGSILGALAFLLLAAITVLAVVFQRKRRGTGYTEQLQQYSSPGLGVKYYIDPSTYEDPCQAIRELAREVDPAYIKIEEVIGTGSFGEVRQGRLQPRGRREQTVAIQALWAGGAESLQMTFLGRAAVLGQFQHPNILRLEGVVTKSRPLMVLTEFMELGPLDSFLRQREGQFSSLQLVAMQRGVAAAMQYLSSFAFVHRSLSAHSVLVNSHLVCKVARLGHSPQGPSCLLRWAAPEVIAHGKHTTSSDVWSFGILMWEVMSYGERPYWDMSEQEVLNAIEQEFRLPPPPGCPPGLHLLMLDTWQKDRARRPHFDQLVAAFDKMIRKPDTLQAGGDPGERPSQALLTPVALDFPCLDSPQAWLSAIGLECYQDNFSKFGLCTFSDVAQLSLEDLPALGITLAGHQKKLLHHIQLLQQHLRQQGSVEV共1021aa，N-15aa為信號肽6.案例1我們沒有的蛋白質(zhì)（2）WDEVSVLDDQRRLTRTFEAC7.案例1我們沒有的蛋白質(zhì)（3）GENEID:2051EPHB6|EPHreceptorB6[Homosapiens]Query1WDEVSVLDDQRRLTRTFEAC20WDEVSVLDDQRRLTRTFEACSbjct41WDEVSVLDDQRRLTRTFEAC60GENEID:814CAMK4|calcium/calmodulin-dependentproteinkinaseIVQuery1WDEVS----------———-VLDDQRRLTRTFEA19WDEVSVLD++RLTTF+ASbjct69WDEVSLNAKDLVRKLIVLDPKKRLT-TFQA978.案例2家族性蛋白的多肽設(shè)計（transgelin）Thetransgelinfamilyisagroupofproteinsthatbelongto22kdactin-relatedcorpninsuperfamily.Ofallthreeisoforms,transgelin1isthebestcharacterized.Transgelin1,alsoknownasSM22alpha,isaspecificmarkerfordifferentiatedsmoothmusclecells.Transgelin2,alsoknownasSM22beta,isexpressedbybothsmoothmuscleandnon-smoothmusclecellsinatemporallyandspatiallyregulatedpattern.Trangenlin3,alsoknownasNP25,isonlyfoundinhighlydifferentiatedneuronalcells.9.案例2家族性蛋白的多肽設(shè)計（transgelin）210.案例2家族性蛋白的多肽設(shè)計（transgelin）3Transgelin1:KYDEELEERLVEWIIVQCTransgelin2:QPGRENFQNWLKDGTVLC11.案例3家族性蛋白的多肽設(shè)計RAB3A,3B,3CSequence2:NM_002866,RAB3A,memberRASoncogenefamilySequence3:NM_002867,RAB3B,memberRASoncogenefamilySequence4:NM_138453,RAB3C,memberRASoncogenefamily12.案例4家族性蛋白的多肽設(shè)計actin對應(yīng)的處理方法？13.案例5同一蛋白質(zhì)不同亞型的多肽設(shè)計NFKB1NFKB1,

NUCLEARFACTORKAPPA-B,SUBUNIT1;NFKB1Alternativetitles;symbolsTRANSCRIPTIONFACTORNFKB1

NUCLEARFACTOROFKAPPALIGHTCHAINGENEENHANCERINBCELLS1

NFKBp105,INCLUDED

NFKBp50,INCLUDED14.案例5同一蛋白質(zhì)不同亞型的多肽設(shè)計NFKB1Thisgeneencodesa105kDproteinwhichcanundergocotranslationalprocessingbythe26Sproteasometoproducea50kDprotein.The105kDproteinisaRelprotein-specifictranscriptioninhibitorandthe50kDproteinisaDNAbindingsubunitoftheNF-kappa-B(NFKB)proteincomplex.1

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433aminoacidsoffulllength968p105proteinformp50.15.案例5同一蛋白質(zhì)不同亞型的多肽設(shè)計NFKB1MAEDDPYLGRPEQMFHLDPSLTHTIFNPEVFQPQMALPTADGPYLQILEQPKQRGFRFRYVCEGPSHGGLPGASSEKNKKSYPQVKICNYVGPAKVIVQLVTNGKNIHLHAHSLVGKHCEDGICTVTAGPKDMVVGFANLGILHVTKKKVFETLEARMTEACIRGYNPGLLVHPDLAYLQAEGGGDRQLGDREKELIRQAALQQTKEMDLSVVRLMFTAFLPDSTGSFTRRLEPVVSDAIYDSKAPNASNLKIVRMDRTAGCVTGGEEIYLLCDKVQKDDIQIRFYEEEENGGVWEGFGDFSPTDVHRQFAIVFKTPKYKDINITKPASVFVQLRRKSDLETSEPKPFLYYPEIKDKEEVQRKRQKLMPNFSDSFGGGSGAGAGGGGMFGSGGGGGGTGSTGPGYSFPHYGFPTYGGITFHPGTTKSNAGMKHGTMDTESKKDPEGCDKSDDKNTVNLFGKVIETTEQDQEPSEATVGNGEVTLTYATGTKEESAGVQDNLFLEKAMQLAKRHANALFDYAVTGDVKMLLAVQRHLTAVQDENGDSVLHLAIIHLHSQLVRDLLEVTSGLISDDIINMRNDLYQTPLHLAVITKQEDVVEDLLRAGADLSLLDRLGNSVLHLAAKEGHDKVLSILLKHKKAALLLDHPNGDGLNAIHLAMMSNSLPCLLLLVAAGADVNAQEQKSGRTALHLAVEHDNISLAGCLLLEGDAHVDSTTYDGTTPLHIAAGRGSTRLAALLKAAGADPLVENFEPLYDLDDSWENAGEDEGVVPGTTPLDMATSWQVFDILNGKPYEPEFTSDDLLAQGDMKQLAEDVKLQLYKLLEIPDPDKNWATLAQKLGLGILNNAFRLSPAPSKTLMDNYEVSGGTVRELVEALRQMGYTEAIEVIQAASSPVKTTSQAHSLPLSPASTRQQIDELRDSDSVCDSGVETSFRKLSFTESLTSGASLLTLNKMPHDYGQEGPLEGKI16.案例5同一蛋白質(zhì)不同亞型的多肽設(shè)計NFKB117.案例6同一蛋白質(zhì)不同亞型的多肽設(shè)計CXCR3TheCXCR3proteinbelongstoGprotein-coupledreceptorfamily.Severalalternativelysplicedtranscriptvariantsencodingdistinctisoformshavebeenreported.TheCXCR3Bproteincontains416aminoacidsandhas