...
首页> 外文期刊>Analytical methods >A simple and efficient approach to improve protein identification by the peptide mass fingerprinting method: concomitant use of negative ionization
【24h】

A simple and efficient approach to improve protein identification by the peptide mass fingerprinting method: concomitant use of negative ionization

机译:一种简单有效的方法,可通过肽质量指纹法改善蛋白质鉴定:伴随使用负离子化

获取原文
   

获取外文期刊封面封底 >>

       

摘要

Peptidemassfingerprinting(PMF)hasbeenwidelyusedasanefficientanalyticalstrategyforproteinidentification.Thisismostcommonlyusedwithacombinationofproteindigestionusingsequence-specificproteasesandMALDI-TOFMS.Thendatabasesearchesareperformedcomparingthepatternoftheexperimentallyobtainedmasseswiththepatternofthetheoreticalpeptidemassesofproteinsstoredinthedatabase.ThepositiveionizationmodehasbeenmainlyusedforMALDIanalyseswithafewexceptionsforphosphopeptides,oligonucleotides,etc.Therefore,nonphosphorylatedpeptidesthathavelowpIvaluescouldbemissedfromPMFusingthepositiveionizationmode.Here,weintroduceoptimalconditionsfornegativeionizationofpeptidesandthepracticaladvantagesofnegativeionizationsinPMF.AngiotensinI(pI6.9)andbovineserumalbumin(BSA)trypticdigestswereusedasmodelpeptides.Eightmatrixcandidatesandsevenadditiveswereexaminedintermsofsensitivity,robustnessandreproducibility.ThecombinationofDHBandphosphoricacidwasthebestconditionfornegativeionizationofpeptidesandwasfoundtobecompatiblewiththepositiveionizationmode.Using150mMDHB/1%phosphoricacid,thecoverage(%byaminoacidcount)ofBSAtrypticdigest(0.6pmolperspot)totaled67.2%(negative+positive).The24.1%ofpeptides(pIrange4.1–6.2)weredetectedonlybynegativeionization,whichindicatedthatacidicpeptideswereefficientlyrecoveredbythenegativeionmode.Thismethodologyhasbeensuccessfullyemployedtoanalyzeotherproteinswithoutfalsepositiveidentifications...
机译:Peptidemassfingerprinting(PMF)hasbeenwidelyusedasanefficientanalyticalstrategyforproteinidentification.Thisismostcommonlyusedwithacombinationofproteindigestionusingsequence-specificproteasesandMALDI-TOFMS.Thendatabasesearchesareperformedcomparingthepatternoftheexperimentallyobtainedmasseswiththepatternofthetheoreticalpeptidemassesofproteinsstoredinthedatabase.ThepositiveionizationmodehasbeenmainlyusedforMALDIanalyseswithafewexceptionsforphosphopeptides,寡核苷酸,etc.Therefore,nonphosphorylatedpeptidesthathavelowpIvaluescouldbemissedfromPMFusingthepositiveionizationmode.Here,weintroduceoptimalconditionsfornegativeionizationofpeptidesandthepracticaladvantagesofnegativeionizationsinPMF.AngiotensinI(pI6.9)andbovineserumalbumin(BSA)trypticdigestswereusedasmodelpeptides.Eightmatrixcandidatesandsevenadditiveswereexaminedintermsofsensitivity,robustnessandreproducibility.ThecombinationofDHBandphosphoricacidwasthebestconditionfornegativeionizationofpeptidesandwasfoundtobecompatiblewiththe正离子化模式。使用150mMDHB / 1%磷酸,BSA胰消化物(0.6pmolperspot)的覆盖率(%pmolperpot)占67.2%(负+阳性)。只有2个负电性被负离子鉴定,表明其是负性的,这表明肽被否定地还原。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱:kefu@zhangqiaokeyan.com

京公网安备:11010802029741号 ICP备案号:京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有
  • 客服微信

  • 服务号