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ID:41116441
大小:3.38 MB
页数:214页
时间:2019-08-16
《Chapter 18 Preparation and Handling of Peptides》由会员上传分享,免费在线阅读,更多相关内容在学术论文-天天文库。
1、CHAPTER18PreparationandHandlingofPeptidesINTRODUCTIONAnimportantcomponentofthearmamentariumofmodernproteinscientistsistheabilitytodesignandconstructpeptidesofalmostanysequence.Peptidesmaybeemployedforawiderangeofapplications,includingassubstratesforkinases,proteases,orglycosidases,orasantigensto
2、produceantiserathatmayrecognizeaproteincontainingthatsequence.Peptidesalsohavemanybiologicalfunctionsoftheirown,interactingwithreceptorstostimulatechangesincellularfunction,oralteringtheinteractionsbetweencells,tociteonlyafewexamples.Chemistsdevelopedthemeanstosynthesizepeptidesearlyinthiscentur
3、y(UNIT18.1).Theabilitytocreateamoleculethatpossessedbiologicalactivitybyorganicsynthesisiscertainlyamilestoneinourscience.Formanyyears,however,peptidesynthesiswaspracticedonlybyhighlyskilledanddedicatedindividuals,andthelengthytimerequiredtoobtainevensimplepeptideslimitedtheiruseasreagentsforbio
4、logicalexperiments.Difficultiesinpurificationeffectivelyeliminatedthestudyofcomplexpeptidesofmorethan5to10aminoacids.Severaldevelopmentshaverevolutionizedthisfield,fortunately,andtheyhavebroughtpeptidescienceintotherealmofstandardbiologicaltechniquesavailabletomanyscien-tists.Firstwasthedevelopment
5、ofsolid-phasemethodsbyBruceMerrifieldandsubse-quentimprovementsinthereagentsavailableforsynthesisthataredetailedinUNIT18.1.Secondwasthedesignandconstructionofautomatedinstrumentstoperformtherepet-itivestepsofthesolid-phasemethodology,culminatinginthe1980swithinstrumentsthatprovidednearlyglobalacc
6、essibilitytosyntheticpeptidesofever-increasinglength.Anotheradvancewastheoptimizationofreversed-phaseHPLCasaseparationmethodfortherapidpurificationandanalysisofsyntheticpeptides(UNIT11.6).Afutureunitinthischapterwillfocusonmedium-to-largescalepeptidepurificationbyHPLCmethods.Inaddition,recentadvan
7、cesinmassspectroscopy(Chapter16)haveprovidedanothercriticalanalyticaltooltoverifytheproductsofsynthesis.Aconvenient,low-costmethodforcreatinglargenumbersofrelatedpeptides,orforassemblyofasetofpeptidesthattogetherspananentire
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