第十八讲 朱冰 Protein Protein Interaction

《第十八讲 朱冰 Protein Protein Interaction》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、Protein-ProteinInteraction&ItsApplicationsInEpigeneticStudies朱冰2017中国科学院大学研究生课WhytoStudyProtein-ProteinInteraction?•Proteinsmightformastablecomplex,participatinginsamebiologicalprocesses•Proteinsmightregulatethebiologicalactivitiesofotherproteins•Proteinsmightserveastheenzymesorsubs

2、tratesforotherproteins•Proteinsmightbindotherproteins,facilitatingdownstreambiologicaleventsTechnologiesforstudyingprotein-proteininteractionsSTOP!KnowYourAimsFirstAimsofYourResearchLookingforUnknownInteraction?VerifyingaCandidateInteraction?WhatKindofInteraction?TwoCategoriesofProt

3、ein-ProteinInteractionStableComplexTransientInteraction•Subunitsremainina•Interactionoccursonlyatcomplexatmosttimeparticularmoments•Subunitsare•Interactionpartnersexistinastoichiometricalsub-stoichiometricalmanner•Interactioncanwithstand•InteractioncanNOTwithstandstringentconditions

4、,suchstringentconditions,suchasashighsaltanddetergenthighsaltanddetergentExamplesofVariousProtein-ProteinInteractions(1)EZH2,EED,SUZ12,RbAp48,AEBP2formsahistoneH3K27methylasecomplexCaoetal.Science2002298:1039ExamplesofVariousProtein-ProteinInteractions(2)RNAPolIIcoreenzymeisaprotein

5、complexVariousGTFsinteractwithPolIIatparticularstagesoftranscriptiontofacilitatetranscriptionHampseyM&ReinbergD.EncyclopediaofLifeScience2001ExamplesofVariousProtein-ProteinInteractions(3)ProductE+S<--->(ES<--->ES*<--->EP)<--->E+PEnzyme-SubstrateInteractionsAreTransientExamplesofVar

6、iousProtein-ProteinInteractions(4)Proteinsmightbindtootherproteins,facilitatingdownstreameventsBannisterAJetal.Nature2001410:120Technologiesforstudyingprotein-proteininteractionsMolecularBiologybasedtechnologiesYeasttwohybridsystemBiochemistrybasedtechnologiesConventionalpurificatio

7、nAffinitypurificationCo-IP&pulldownAffinitypurificationassistedwithSILACFluorescencebasetechnologiesImmuno-fluorescencebasedco-localizationBimolecularfluorescencecomplementationassay(BiFC)Fluorescenceresonanceenergytransfer(FRET)BiophysicsbasedtechnologiesSurfacePlasmonResonanceIsot

8、hermalTitrationCalo