氨基酸多肽和蛋白质ppt课件.ppt

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1、Chapter3AminoAcids,Peptides,andProteins3.2.6DetreminationoftheaminoacidsequencesofproteinsSangerworkedoutthefirstaminoacidsequenceofapeptide(bovineinsulin)in1953.AminoAcidsSequenceAnalysisStrategyforsequenceanalysis：AminoAcidsSequenceAnalysisAminoAcidsSequenceAn

2、alysisAminoAcidsSequenceAnalysisSSAminoAcidsSequenceAnalysis。。。。。。。。。。AminoAcidsSequenceAnalysisAminoAcidsSequenceAnalysisGAminoAcidsSequenceAnalysisAminoAcidsSequenceAnalysisGIVAminoAcidsSequenceAnalysisGIVEQCCTSIENYCNCSLYQLAminoAcidsSequenceAnalysisGIVEQCCTSIEN

3、YCNCSLYQLGIVEQCCTSICSLYQLENYCNAminoAcidsSequenceAnalysisGIVEQCCTSTENYCNCSLYQLGIVEQCCTSICSLYQLENYCNAminoAcidsSequenceAnalysisAminoAcidsSequenceAnalysisAminoacidsequenceofinsulinAminoAcidsSequenceAnalysisMethodforsequenceanalysis：AminoAcidsSequenceAnalysisDetermina

4、tionoftheaminoacidcompositionoftheproteinHydrolysisofproteinSeparationofaminoacidsIdentificationandquantificationofaminoacidsAminoAcidsSequenceAnalysisHydrolysisofproteins：HydrolysisbyacidHydrolysisbybaseHydrolysisbyenzymeFullhydrolysisPartialhydrolysisAminoAcids

5、AminoAcidsIon-exchangechromatographyAminoAcidsSequenceAnalysisIon-exchangechromatographyAminoAcidsSequenceAnalysisAminoacidsequenceanalyzer:ion-exchangechromatography+colorreagents+spectrophotometerAminoAcidsSequenceAnalysisIdentificationoftheN-terminalandC-termi

6、nalaminoacidDNFBmethodDNSmethodPITCmethodAminopeptidasemethodHydrazinolysisCarboxypeptidaseN-termianlC-termianlAminoAcidsSequenceAnalysisDNS-DansylchlorideAminoAcidsSequenceAnalysisAminoAcidsSequenceAnalysisTheaminoacidsequenceofashortpeptidecanbeefficientlydeter

7、minedbyEdmandegradation.Theunchargedterminalaminogroupisreactedwithphenylisothiocyanatetoformaphenylthiocarbamylpeptide.TheN-terminalaminoacidresidueisliberatedasacyclicphenylthiohydantoin(PTH)derivativeundermildlyacidconditions,leavingtherestofthepeptidechainint

8、act.ThePTHderivative(thustheaminoacidresidue)canbeidentifiedbychromatographicmethodsThenewlyexposedN-terminalaminoacidresiduecanbeidentifiedbyrepeatingtheabove