GDP Release Preferentially Occurs on the Phosphate Side in Heterotrimeric G-proteins 英文参考文献.docVIP

GDPReleasePreferentiallyOccursonthePhosphateSide inHeterotrimericG-proteins MaximeLouet,JeanMartinez,NicolasFloquet* InstitutdesBiomole′cules MaxMousseron(IBMM),CNRSUMR5247,Universite′ Montpellier1,Universite′ Montpellier2,Faculte′ dePharmacie,Montpellier,France Abstract Afterextra-cellularstimulationofG-ProteinCoupledReceptors(GPCRs),GDP/GTPexchangeappearsasthekey,ratelimiting step of the intracellular activation cycle of heterotrimeric G-proteins. Despite the availability of a large number of X-ray structures, the mechanism ofGDP release out ofheterotrimeric G-proteins stillremains unknown at themolecular level. Starting from the available X-ray structure, extensive unconstrained/constrained molecular dynamics simulations were performed on the complete membrane-anchored Gi heterotrimer complexed to GDP, for a total simulation time overcoming500ns.BycombiningTargetedMolecularDynamics(TMD)andfreeenergyprofilesreconstructionbyumbrella sampling,ourdatasuggestthatthereleaseofGDPwasmuchmorefavoredonitsphosphateside.Interestingly,uponthe forcedextractionofGDPonthisside,thewholeproteinencounteredlarge,collectivemotionsinperfectagreementwith thosewedescribedpreviouslyincludingadomaintodomainmotionbetweenthetworas-likeandhelicalsub-domainsof Ga. Citation:LouetM,MartinezJ,FloquetN(2012)GDPReleasePreferentiallyOccursonthePhosphateSideinHeterotrimericG-proteins.PLoSComputBiol8(7): e1002595.doi:10.1371/journal.pcbi.1002595 Editor:JamesM.Briggs,UniversityofHouston,UnitedStatesofAmerica ReceivedFebruary7,2012;AcceptedMay21,2012;PublishedJuly19,2012 Copyright: ? 2012 Louet et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricteduse,distribution,andreproductioninanymedium,providedtheoriginalauthorandsourcearecredited. Funding: This work was fundedbyAgence Nationale dela Recherche (ANR) (Programme blanc: ‘‘GHSR-Sig’’). The fundershadnorole in study design, data collectionandanalysis,decisiontopub