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1、2009年3月色谱Vo.l27No.2March2009ChineseJournalofChromatography158~163磺化修饰结合离子交换色谱和生物质谱富集鉴定含组氨酸肽段1,222212*曹冬,周春喜,张养军,韩春光,邓玉林,钱小红(1.北京理工大学,北京100081;2.北京蛋白质组研究中心蛋白质组学国家重点实验室北京放射与辐射医学研究所,北京102206)摘要:通过在肽段的N端引入磺酸基,从而使含组氨酸的肽段与其他肽段在pH<30的条件下产生电荷差异,建立了一种基于强阳离子交换色谱(SCX)结合生物质谱富集鉴定含组氨酸肽段的方法,并以
2、含有组氨酸的标准蛋白质为模型,进行了方法学考察。结果表明,经N端磺酸化后,含组氨酸的肽段能有效地被阳离子交换色谱富集,且在肽的N端引入磺酸基促进了肽的裂解,使之产生简单而信息丰富的二级质谱图,从而得到完美的质谱鉴定结果。这说明磺化修饰结合强阳离子交换色谱用于含组氨酸肽段的富集鉴定是可行的,且具有在蛋白质组研究中应用的潜力。关键词:强阳离子交换色谱;基质辅助激光解吸电离飞行时间串联质谱;磺化修饰;富集;鉴定;组氨酸标签肽中图分类号:O658文献标识码:A文章编号:10008713(2009)02015806栏目类别:研究论文Sulfonationmo
3、dificationassistedenrichmentandidentificationofhistidinecontainingpeptidesbystrongcationexchangechromatographyandmassspectrometry1,2222CAODong,ZHOUChunxi,ZHANGYangjun,HANChunguang,12*DENGYulin,QIANXiaohong(1.BeijingInstituteofTechnology,Beijing100081,China;2.StateKeyLaboratoryofProte
4、omics,BeijingProteomeResearchCenter,BeijingInstituteofRadiationMedicine,Beijing102206,China)Abstract:BythesulfonationattheNterminalofpeptides,thechargestateofhistidinecontainingpeptidesisdifferentfromthatofotherpeptidesinpH<30solution.Basedonthisdifference,anewmethodwasdevelopedto
5、isolateandidentifysulfonatedhistidinecontainingpeptidesfromtrypticdigestofproteinsbystrongcationexchange(SCX)chromatographyandmatrixassistedlaserdesorption/ionizationtandemtimeofflightmassspectrometry(MALDITOFMS/MS).Usingthestandardproteinscontaininghistidinesasthemode,lthemetho
6、dologywasevaluated.TheresultsshowthatsulfonatedhistidinecontainingpeptideswereefficientlyenrichedbySCX,andtheNterminalsulfonationofthepeptidessimplifiestheinterpretationoftheacquiredmassspectraandfacilitatesthesequencingofhistidinecontainingpeptidesbypro2ducingconsecutiveandpredomi
7、nantionsinpositivemodeMSspectra,whichisthoughttobetheresultofthechargeneutralizationofbionsbytheNterminalsulfonicacidgroup.Thediscriminationofbionsandyionscangreatlyenhancetheconfidenceinpeptideandsubsequentproteinidentification.Itisfeasibletoisolateandenrichthehistidinecontaining
