多肽合成中半胱氨酸检测的简易方法

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1、JournalofPeptideScienceJ.Pept.Sci.2008;14:838844Publishedonline11February2008inWileyInterScience(www.interscience.wiley.com).DOI:10.1002/psc.1012AsimplemethodformonitoringthecysteinecontentinsyntheticpeptidesKATAHORVATI,SZILVIAB´OSZE,FERENCHUDECZandHEDVIGMEDZIHRADSZKY-SCHWEIGER*

2、˝ResearchGroupofPeptideChemistry,HungarianAcademyofSciences,H-1117Budapest,Pazm´anyP´eterst.1/A,Hungary´Received31October2007;Revised28November2007;Accepted28November2007Abstract:Anewmethodforthedeterminationofthecysteinecontentofsyntheticpeptidesisdescribed.Forthispurposethecla

3、ssicalaminoacidanalysisisnotsuitable,ascysteinemayundergooxidativedimerisationtocystineduringthehydrolysisorevaporation.TheintactpeptidewasreactedwithN-ethylmaleimide(NEM)yieldingastableS-(N-ethylsuccinimido)-cysteinederivative,whichcouldbeseparatedbyRP-HPLCandcharacterisedbymas

4、sspectrometry.Forthequantitativedeterminationlessthan0.1µMpeptidewassufficient.Copyright2008EuropeanPeptideSocietyandJohnWiley&Sons,Ltd.Keywords:peptide;cysteine;N-ethylmaleimide;HPLC;massspectrometryINTRODUCTIONInthecaseofcysteine-containingpeptidesonehastotakeintoaccountsomedi

5、fficulties.ItiswellknownthatPeptide/polypeptideandprotein-basedconjugateswithcysteineisverysensitivetooxidation.Evenundercare-covalentlyattachedepitopepeptidesareconsideredasfullyselectedreactionconditionscysteineresiduescanpotentialsyntheticvaccineordiagnosticscandidatesbeeasily

6、oxidisedtocystine.Asimilarside-reactioncanandalsofordrugtargetingtobeusedinchemother-occurduringsynthesisaswell,itisnoteasytodecideapy[1].Applicationsofcysteine-insertionandthioetherwhetherthecystinedeterminedbytheaminoacidanal-linkageapproachestothepreparationofanumberysisorigi

7、natedfromthehydrolysateortheinvestigatedofbioactivepeptideconjugatesarereported[13].peptidewascontaminated.Onion-exchangechro-Peptidescontainingepitopesfrom(i)Herpessimplexmatographytheretentiontimeofprolineandcysteineisvirustype1glycoproteinD,(ii)aspecificN-terminalalmostthesame

8、,thereforecysteinecanbedeterminedβ-amyloidepito