您好,欢迎访问三七文档
当前位置:首页 > 电子/通信 > 综合/其它 > 蛋白质中二硫键的定位及其质谱分析
:20077,:200793(No130672600,20675079)33e2mail:syliu19@yahoo.com.cn31,211133(11130022;21100039),,,,:O657163;Q51:A:10052281X(2008)0620975209ProteinDisulfideBondDeterminationandItsAnalysisbyMassSpectrometryQiuXiaoyan1,2CuiMeng1LiuZhiqiang1LiuShuying133(11ChangchunCenterofMassSpectrometry,ChangchunInstituteofAppliedChemistry,ChineseAcademyofSciences,Changchun130022,China;21GraduateSchooloftheChineseAcademyofSciences,Beijing100039,China)AbstractDisulfidebondsareoneofthemostcommoncovalentposttranslationalmodificationsofproteins.Theyplayanimportantroleinmaintainingthethree2dimensionalstructuresofproteinsandtheirbiologicalactivities.Therefore,thedeterminationofdisulfidebondsbecomesanimportantaspectforobtainingacomprehensiveunderstandingofthechemicalstructureofaprotein.Numerousexperimentalmethodshavebeendevelopedforthedeterminationofdisulfidebondsinproteins.Modernmassspectrometryhasdevelopedasanimportanttoolfortheanalysisofdisulfidebondpatternsduetoitsadvantagesofbeingsimple,rapidandsensitive.Someusefulmethodsforassignmentofdisulfidebondsinproteinsareintroduced,thedevelopmentsandapplicationsofmassspectrometryinthisareaarereviewed.Keywordsassignmentofdisulfidebond;massspectrometry;MSPMS;TCEP;stableisotopelabeling1,,[15][6,7][810],,P[11],[1214],X(X2raycrystallography)[15](NMR)[16],,1955Ryle[17,18]AB,A20620086PROGRESSINCHEMISTRYVol.20No.6June,2008,(CysA6,CysA7)1960,Moore[19](RNase)124,4,:(1),;(2);(3);(4);(5),,[1721],,pH[2230],(pepsin)[19,22,24],,,,,,:X[3134];[35],,[26,3641],2X[42],NMR[43],,(diagonalelectrophoresis)[22]:(1);(2)pH=615,;(3)(CHOOOH),,(SO3H),;(4)90,,,,,,,SH,,,(HPLC)[28,4446],HPLC,,[24],Edman(Edmandegradation)[13](aminoacidanalysis)[19,47],,,[18,19,22,28,4446,4851],Edman,(multiple2stepenzymaticorchemicaldigestion),,67920,;,,;,X;,315,,[5258],P,,,,(conformationalenergy),tris(22carboxyethyl)phosphine(TCEP)1993Gray[52]TCEP,,HPLC(iodoacetamide,IAM),Cys,[52,5963],IAM,pH=8[61]1996Wu[53]2225222nitro252thiocyanobenzoicacid(NTCB),N,22242(22iminothiazolidine242carboxylpeptides),MALDI2MS1997Wu[54](1),TCEPpH=3,122422(12cyano242dimethylamino2pyridiniumtetrafluoro2borate,CDAP),[54,6467],pH,1(ITC)[54]Fig.1Principleofthepartialreductionmethod(ITCstandsforiminothiazolidinylcarboxylresidueattheaminoterminus)[54]1molL-1NH4OH,SCNN,NH4OH,Qi[67,114]3(2Cys2Cys2Cys2)CysN,,,2(2mercaptoethanol)(dithiothreitol,DTT),,,(tributylphosphine,TBP)[68],,,,TCEP[69]7796,TBP,TCEP,pH(pH=3)[70,71],TCEPDTT,TCEP[55,72]3[73],80(fastatombombardment,FAB)[74](plasmadesorption,PD)[7578](electrosprayionization,ESI)[82,83](matrix2assistedlaserdesorptionionization,MALDI)[8489],[25,7981][24],pH,Edman(1)MALDI2TOF2MSESI2MS(liquidchromatography,LC)Tetsuya[90]LCPESI2MSendoPG(tandemmassspectrometry,orMSPMS),(2),FAB[91]ESI(ESI2triple2quadrupole2MSPMS)[6,29]MALDI2TOF(post2sourcedecay,PSD)[36,92]2[138]Fig.2Complexdisulfide2linkedpeptides[138]MSPMS[29,36]Mhatre[93]MALDI2TOF,IgG1(mAb)Zhang[94]RP2HPLCPMS,nanoESI2QTOFMSPMSEdmanIgG4mAbYen[37]LCPESI2MSPMSSequestMascot[95]TCEPLCPESI2MSPMS[96],65TCEP(011015mmolL-1),(4)TCEP,N2ethylmaleimide(NEM)maleimide2biotin(M2biotin)n,n-1NEM,M2biotinIAM3,MSPMS(3)NEMM2biotinpH(615),pH,,Schnolzer[97]H218OC118O,LCESI879201Table1Representativeexamplesofdisulfidebondarrangementsinproteindeterminedbymassspectrometryproteinmethodologyref.insulinFAB2MSofpepticdigestpeptides24henandduckegg2whitelysozymesFAB2MSofcombinedCNBrPpepticdigestpeptides80henegg2whitelysozymeandbovineribonucleaseA(RNasA)FAB2MSofpeptidesfromvariousdigestsinvolvingCNBrPtrypsinPdiluteacidPchymotrypsinPendoproteinaseGlu2C25RNaseAFAB2MSofcombinedCNBrPtrypticdigestpeptides26antothrombinFAB2MSofcombinedendoproteinaseGlu2CPtrypticorpepticdigestpeptides28paim1,2amylaseinhibitorFAB2MSofendoproteinaseGlu2Cdigestpeptides47wheatkernel2amylaseinhibitorFAB2MSofdifferentcombinationsofelastasePtrypsinorelastasePtrypsinPchymotrypsinorelastasePtrypsinPpepsinandofsomepeptidesshortenedbymanualEdmandegradation139protozoanEr21andEr22MALDI2TOF2MSandESI2TOF2MSofthermolyticdigestpeptides46echistatinESI2MSandESI2MSPMSofpeptidesderivedbyoxalicacidhydrolysis29bovinedopamine2hydroxylaseESI2MSandMALDI2TOF2MSoftrypticorendoproteinaseGlu2Corpepticdigestpeptides140saposinCandBESI2MSandFAB2MSofcombinedCNBrPpepticPendoproteinaseGlu2CorCNBrPtrypticPendoproteinaseAsp2NPchymotrypticpeptides141brassicanapusseedstorageprotein,napinMALDI2TOF2MSofcombinedendoproteinaseLys2CPthermolyticdigestpeptides142plasmodiumapicalmembraneantigen21IonTrap2MSofcombinedtrypticPthermolyticdigestpeptides143bitisarietansbitistatinMALDI2TOF2MSandESI2MSPMSoftrypticandoxalicaciddigestpeptides144humaninsulinreceptorectodomainMALDI2TOF2MSofcombinedCNBrPtrypticdigestpeptides145humanrespiratorysyncytialvirusattachmentglycoproteinMALDI2TOF2MSandMALDI2TOF2PSD2MSofcombinedtrypticPpepticortrypticPthermolyticdigestpeptides92newcastlediseasevirus(NDV)hemagglutinin2neuraminidase(HN)MALDI2TOF2MSandESI2MSPMSofpepticorpepticPCNBrorpepticPtrypticorpepticPchymotrypticdigestpeptides6humanagouti2relatedproteinMALDI2TOF2MSoftrypticPpepticdigestpeptidesofp
本文标题:蛋白质中二硫键的定位及其质谱分析
链接地址:https://www.777doc.com/doc-5470303 .html