基于生物质谱的蛋白质组学绝对定量方法研究进展

:20070428;:20070723:(2001CB510201,2004CB520802);973(2006CB910803);(30321003204050172050501820505019);(H030230280190):(1980~),(),,,Email:[e-mail]:(1953~),(),,,Email:[e-mail]29320085JournalofChineseMassSpectrometrySocietyVol.29No.3May2008曹冬,张养军,钱小红(,,102206):();;,,:;;;;:O657.63;Q1:A:10042997(2008)0318507DevelopmentofAbsoluteQuantificationofProteomicsBasedonBioMassSpectrumCAODong,ZHANGYangjun,QIANXiaohong(BeijingInstituteofRadiationMedicine,BeijingProteomeResearchCenter,Beijing102206)Abstract:Thequantitativemethodsofproteomicsincludetherelativequantitationandabsolutequantitationofproteins.Relativequantitationofproteomicsistheanalysisoftherelativechangesoftheexpressedproteinsinthesamplesoriginatedfromdifferentphysiologicalandpathologicalsituations.Theabsolutequantitationofproteomicsistodeterminetheabsoluteamountorconcentrationofeachproteininabiologicalsample,suchascells,tissueorbodyfluid.Currently,themainstrategiesofabsolutequantitationofproteomicsarecomposedofthequantitativemethodsbasedoninternalstandardstrategyandthelabelfreetechniquewiththestatisticanalysisofMSdata.Thequantitativeinformationoftheproteomecanhelpusunderstandthebiologicalfunctionofproteinsintheinteractionnetwork.Withthedeterminationoftheabsolutequantitationofthebiomarker,wecandirectlyjudgethestatusofadisease,whichwillbenefittheclinicaldiagnosisandthetreatmentofthedisease.Keywords:quantitativeproteomics;absolutequantitation;isotopiclabeledmethod;labelfreequantitation;biomassspectrum,()[12],,,2090[3],10,:()[45],;[67],;,,,,:(),,(2DE)[8](differentialingelelectrophoresis,DIGE)[9](isotopecodedaffinitytags,ICAT)[1011](isobarictagsforrelativeandabsolutequantitation,iTRAQ)[12][13];,,,4(stoichiometriccore),,,,,[14][15],,,,,,,,,,11.1:(0.9~2ku),(13C15N2H)(),():1);2)Gerber[16],(SRM),,,(LFTGHPETLEKALELFR),(LFTGHPETL*18629EKALEL*FR,L*),SRM,Beynon[17]20,:(Q),,,(QCAT),15N(15NH4Cl);QCAT,,,15NQQ,QCAT:1)1000~2000(),(QCAT),QCAT;2)QCAT,150ku,QCAT,Havlis[18]18O,,18O,,;Mayya[19](LCSRM)4,1.2:,,,,MALDITOFMS,,Mirgorodskaya[20],,,:1);2)MALDI,;3)1.3(ICPMS),,Navaza[21],:BNPP,HPLC,ICPMS,ESIMS,,,ICPMS,BNPP,,,Schaumloffel[22],34S,34S,,,,:1)MS;2);3);4)1873:1.4:,,iTRAQ(IsobaricTagsforRelativeandAbsoluteQuantitation)Ross[23]iTRAQ,iTRAQ,(NHS)13C114115116117,,13C18O,145N,iTRAQ,;,(CO),,114115116117,,iTRAQ,1.5,,Mayr[24],,;,,N,,,,,,,,2(labelfree),,(),,Nakamura[25],MS,MS,,,,,,LCFTMS,,,,Silva[26],,:6,,3,3,,3,(universalsignalresponsefactor),18829Ishihams[27],nanoLCESIMS/MS,,,,,,,mRNADNA,Liu[28],spectralcount,,,2Lu[29](APEX),spectralcounts,,westernblotting2DEmRNAState[30],32DE,,,,,,,,,,,,,,,,:[1]VENTERJC,ADAMSMD,MYERSEW,etal.Thesequenceofhumangenome[J].Science,2001,291(5507):13041351.[2]LANDERES,LINTONLM,BIRRENB,etal.Initialsequencingandanalysisofthehumangenome[J].Nature,2001,409(6822):860921.[3]WASINGERVC,CORDWELLSJ,CERPAPOLJAKA,etal.ProgresswithgeneproductmappingoftheMollicutes:Mycoplasmagenitalium[J].Electrophoresis,1995,16:10901094.[4]LIUH,LIND,YATESJR.Multidimensionalseparationsforprotein/peptideanalysisinthepostgenomicera[J].Biotechniques,2002,32(4):898902.[5]WASHBURNMP,WOLTERSD,YATESJR.Largescaleanalysisoftheyeastproteomebymultidimensionalproteinidentificationtechnology[J].NatBiotech,2001,19:242247.[6]JONSSONAP.Massspectrometryforproteinandpeptidecharacterization[J].CellMolLifeSci,2001,58:868884.[7]YERGEYAL,COORSSENJR,BACKLUNDPS,etal.DenovosequencingofpeptidesusingMALDI/TOFTOF[J].JAmSocMassSpectrom,2002,13(7):748791.[8]LILLEYKS,RAZZAQA,DUPREEP.Twodimensionalgelelectrophoresis:recentadvancesinsamplepreparation,detectionandquantitation[J].CurrentOpinioninChemicalBiology,2002,6(1):4650.[9]ALBANA,DAVIDSO,BJORKESTENL,etal.Anovelexperimentaldesignforcomparativetwodimensionalgelanalysis:twodimensionaldifferencegelelectrophoresisincorporatingapooledinternalstandard[J].Proteomics,2003,3(1):3644.[10]GYGISP,RISTB,GERBERSA,etal.Quantitativeanalysisofcomplexproteinmixturesu1893:singisotopecodedaffinitytags[J].Naturebiotechnology,1999,17(10):994999.[11],,,.[J].,2006,34(7):899904.[12]CHONGPK,GANCS,PHAMTK,etal.Isobarictagsforrelativeandabsolutequantitation(iTRAQ)reproducibility:Implicationofmultipleinjections[J].JournalofProteomeResearch,2006,5(5):12321240.[13]LIUH,ZHANGY,WANGJ,etal.Methodforquantitativeproteomicsresearchbyusingmetalelementchelatedtagscoupledwithmassspectrometry[J].AnalChem,2006,78(18):66146621.[14]HAVLISJ,SHEVCHENKOA.Absolutequantificationofproteinsinsolutionsandinpolyacrylamidegelsbymassspectrometry[J].AnalChem,2004,76(11):30293036.[15]BARNIDGEDR,GOODMANSONMK,KLEEGG,etal.AbsolutequantificationofthemodelbiomarkerprostatespecificantigeninserumbyLCMS/MSusingproteincleavageandisotopedilutionmassspectrometry[J].JournalofProteomeResearch,2004,3(3):644652.[16]GERBERSA,RUSHJ,STEMMANO,etal.AbsolutequantificationofproteinsandphosphoproteinsfromcelllysatesbytandemMS[J].PNAS,2003,100(12):69406945.[17]BEYNONRJ,DOHERTYMK,PRATTJM,etal.MultiplexedabsolutequantificationinproteomicsusingartificialQCATproteinsofconcatenatedsignaturepeptides[J].NatureMethods,2005,2(8):587589