Parkin Mediates Apparent E2-Independent Monoubiquitination In Vitro and Contains an Intrinsic Activity That Catalyzes Polyubiquitination 英文参考文献.docVIP

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Parkin Mediates Apparent E2-Independent Monoubiquitination In Vitro and Contains an Intrinsic Activity That Catalyzes Polyubiquitination 英文参考文献.doc

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Parkin Mediates Apparent E2-Independent Monoubiquitination In Vitro and Contains an Intrinsic Activity That Catalyzes Polyubiquitination 英文参考文献

ParkinMediatesApparentE2-Independent MonoubiquitinationInVitroandContainsanIntrinsic ActivityThatCatalyzesPolyubiquitination KatherineC.M.Chew1.,NoriyukiMatsuda2.,KeikoSaisho2,GraceG.Y.Lim1,ChouChai3,Hui-MeiTan4, KeijiTanaka2,Kah-LeongLim1,3,4* 1DepartmentofPhysiology,NationalUniversityofSingapore,Singapore,Singapore,2LaboratoryofFrontierScience,TokyoMetropolitanInstituteofMedicalScience, Tokyo,Japan,3A*STARDuke-NUSNeuroscienceResearchPartnership,Singapore,Singapore,4NationalNeuroscienceInstitute,Singapore,Singapore Abstract Background:Mutationsintheparkingene,whichencodesaubiquitinligase(E3),areamajorcauseofautosomalrecessive parkinsonism.Althoughparkin-mediatedubiquitinationwasinitiallylinkedtoproteindegradation,accumulatingevidence suggeststhattheenzymeiscapableofcatalyzingmultipleformsofubiquitinmodificationsincludingmonoubiquitination, K48-andK63-linkedpolyubiquitination.Inthisstudy,wesoughttounderstandhowasingleenzymecouldexhibitsuch multifunctionalcatalyticproperties. Methods and Findings: By means of in vitro ubiquitination assays coupled with mass spectrometry analysis, we were surprised to find that parkin is apparently capable of mediating E2-independent protein ubiquitination in vitro , an unprecedented activity exhibited by an E3 member. Interestingly, whereas full length parkin catalyzes solely monoubiquitinationregardlessofthepresenceorabsenceofE2,atruncatedparkinmutantcontainingonlythecatalytic moietysupportsbothE2-independentandE2-dependentassemblyofubiquitinchains. Conclusions: Our results here suggest a complex regulation of parkin’s activity and may help to explain how a single enzymelikeparkincouldmediatediverseformsofubiquitination. Citation:ChewKCM,MatsudaN,SaishoK,LimGGY,ChaiC,etal.(2011)ParkinMediatesApparentE2-IndependentMonoubiquitinationInVitroandContainsan IntrinsicActivityThatCatalyzesPolyubiquitination.PLoSONE6(5):e19720.doi:10.1371/journal.pone.0019720 Editor:HarmKampinga,UniversityofGroningen,TheNetherlands ReceivedDecember13,20