Proteomic-Based Identification of CD4-Interacting Proteins in Human Primary Macrophages 英文参考文献.docVIP

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Proteomic-Based Identification of CD4-Interacting Proteins in Human Primary Macrophages 英文参考文献.doc

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Proteomic-Based Identification of CD4-Interacting Proteins in Human Primary Macrophages 英文参考文献

Proteomic-BasedIdentificationofCD4-Interacting ProteinsinHumanPrimaryMacrophages RuiAndre′ SaraivaRaposo1,2*¤,BenjaminThomas1,3,GabrielaRidlova1,3,WilliamJames1 1SirWilliamDunnSchoolofPathology,UniversityofOxford,Oxford,UnitedKingdom,2GraduatePrograminAreasofBasicandAppliedBiology(GABBA),Universityof Porto,Porto,Portugal,3CentralProteomicsFacility,SirWilliamDunnSchoolofPathology,UniversityofOxford,Oxford,UnitedKingdom Abstract Background:Humanmacrophages(Mw)expresslowlevelsofCD4glycoprotein,whichisconstitutivelyrecycled,and40– 50% of its localization is intracellular at steady-state. Although CD4-interacting proteins in lymphoid cells are well characterised,littleisknownabouttheCD4proteininteraction-networkinhumanMw,whichnotablylackLCK,aSrcfamily proteintyrosinekinasebelievedtostabiliseCD4atthesurfaceofTcells.AsCD4isthemaincellularreceptorusedbyHIV-1, knowledgeofitsmolecularinteractionsisimportantfortheunderstandingofviralinfectionstrategies. Methodology/PrincipalFindings:Weperformedlarge-scaleanti-CD4immunoprecipitationsinhumanprimaryMwfollowed by high-resolution mass spectrometry analysis to elucidate the protein interaction-network involved in induced CD4 internalizationanddegradation.ProteomicanalysisofCD4co-immunoisolatesinrestingMwshowedCD4associationwitha rangeof proteins foundin thecellular cortex, membranerafts andcomponents of clathrin-adaptor proteins, whereas in inducedinternalizationanddegradationCD4isassociatedwithcomponentsofspecificsignaltransduction,transportand theproteasome. Conclusions/Significance:Thisisthefirsttimethattheanti-CD4co-immunoprecipitationsub-proteomehasbeenanalysed inhumanprimaryMw.OurdatahaveidentifiedimportantMwcellsurfaceCD4-interactingproteins,aswellasregulatory proteinsinvolvedininternalizationanddegradation.Thedatagivevaluableinsightsintothemolecularpathwaysinvolved intheregulationofCD4expressioninMwandprovidecandidates/targetsforfurtherbiochemicalstudies. Citation:RaposoRAS,ThomasB,RidlovaG,JamesW(2011)Proteomic-BasedIdent