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医学新知导论PPT课件.ppt

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医学新知导论OutlineTheFiveMassSpectrometryNobelPrizePioneersWHATISA“MASSSPECTROMETER”...?Theblackboxproblem…...“Amassspectrometermeasuresthemolecularweight….”DataProcessinghnHPLC-MSElectro-spray,ESI(電灑法)HPLC-MSMALDI-TOFspectraofapomyoglobinHPLC-MSHPLC-MSESIandAPCIdifferin…HPLC-MSHPLC-MSMALDITime-of-flight(TOF)MSIonTrapFourier-transformIonCyclotronResonance(FT-ICR)SingleMSAnalyzerHPLC-MS/MSTripleQuadrupoleMS/MSScanFunctionsProductIonScanningParkedonPrecursorionPrecursorIonScanningProteomics Introduction Massspectrometryinproteomics Proteinidentification Quantitativeproteomics Post-translationalmodificationProteome:In1993,theterm“Proteome”,byMarcWilkinsandKeithWilliams,wasreferredtothesystematicidentificationoftheentireproteinpopulationexpressedbyagenomeorbyacellortissuetype. Proteomics:Thesubjectofproteomicanalysisoftheproteome(PROTEincomplementexpressedbyagenOMEorbyacellortissuetype).DefinitionofProteomics蛋白體的發展里程碑Whyanalyzegeneexpressionattheproteinlevel?YApolypeptidecanfoldtogenerateaparticularthree-dimensionalstructurespecifiedbyitsaminoacidsequence. Thestructuraldescriptionofproteinsisdescribedintermsoffourlevelsoforganization.MS-basedProteomicsThegeneralPropertyDifferencesbetweenDNAandProteinGoalofGenomicsandProteomicsProteinExtractExcisespot; wash;digestPeptide-MassFingerprinting(PMF)Advantages&Disadvantagesof2DgelApproachLimitationof2-DGelBasedProteomicsPlatformProteinDetectionMethodsHigh-PerformanceLiquidChromatographyMultiDimensionalLiquidChromatographySizeExclusionAffinityChromatographyIonExchangeChromatographyIonExchangeChromatographyHydrophobicInteractionGeneralapproachesProteinIdentificationbyPeptideFragmentFingerprintingSample IonsMALDI-TOF/TOFnanoRPLC-ESI-TandemMSN-andC-terminalPeptidesTerminalpeptidesandiontypesN-andC-terminalPeptidesN-andC-terminalPeptidesN-andC-terminalPeptidesN-andC-terminalPeptidesMassSpectraProteinIdentificationwithMS/MSSequencefromIDMascotSearchParametersMascotSearchResultQuantitativeproteomicsisobaricTagforRelativeandAbsoluteQuanti