蛋白质复合物的纯化

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1、Purificationof蛋白质复合物的纯化MultiproteinComplexesPurificationofmultiproteincomplexes蛋白质复合物的纯化Outline内容•Introduction•简介–蛋白质-蛋白质相互作用–Protein-proteinInteraction–Challengesinpurification–纯化的困难–Analyticaltechniques–分析技术•Purification•纯化–“ngtoµg”规模–“ngtoµg”scale–“µgtomg”规模–“µgtomg”sca

2、le•Summary•结论Introduction简介1/GE/Protein-proteininteractions简介蛋白质-蛋白质复合物Introduction•5000interactionshavebeenmappedinC.elegans•线虫中找出5000相互作用•Understandingthefunctionsofcomplexesand•了解复合物和相互作用的功能可能导致药物的发现interactionswouldleadtoimportantdrugtargets质膜液泡网状组织内皮线粒体细胞质小泡Protein-pr

3、oteininteractionsinC.elegans,Numberofproteinsinvolvedininteractions线虫蛋白质－蛋白质相互作用,酵母每个细胞涉及相互作用的蛋白质数目“Interactome”percellcompartmentsinyeast“Interactome”Li,S.etal.(2004)Science303,540-543Schwikowskiet.al.(2000)NatureBiotech.Vol18,p1257Li,S.etal.(2004)Science303,540-543Schwik

4、owskiet.al.(2000)NatureBiotech.Vol18,p12577/8/GEMultiproteinComplexes/GEMultiproteinComplexes/2008-4-32008-4-3Multiproteincomplexes简介蛋白质复合物Introduction•Assemblyofmorethantwoproteins,•多于两个蛋白质的组合–10to100proteins–10-100蛋白质•Executethevastmajorityofcellularfunctions•执行主要的细胞功能–s

5、ignaltransduction,cellcycleregulation,transcription…–信号传导,细胞周期调控,转译…•2/3ofproteinsinprokaryotes•2/3原核细胞的蛋白质和4/5真核细胞的蛋白质以复and4/5ineukaryotesexist合物形式存在ascomplexes•稳定的和短暂的相互作用•StableandtransientinteractionsAmodeloftranscriptionmediatorcomplexCAmodeloftranscriptionmediatorcom

6、plexCfromSaccharomycescerevisiaefromSaccharomycescerevisiae9/10/GEMultiproteinComplexes/GEMultiproteinComplexes/2008-4-32008-4-3Multiproteincomplexesandfunctions简介蛋白质复合物和功能IntroductionSplicing–removalSpliceosome剪接–去除内含子Intron剪接体SpliceosomeonintronsProteindegradationProteas

7、ome蛋白质降解蛋白酶体ProteasomeRNAsynthesisandExosomecomplexRNA合成和调控外切酶体复合物Exosomecomplexregulation染色质组装ChromatinassemblyNucleosome核小体NucleosomeChromatinassembly多聚腺苷复合物PolyadenylationcomplexRNAPolyadenylationcomplexRNA•Ribosomes•核糖体Ribosomes•Chaperonins•伴侣蛋白Chaperonins11/12/GEMulti

8、proteinComplexes/GEMultiproteinComplexes/2008-4-32008-4-32/GE/Overview:Purificationofcomp