2016
FRAP to Characterize Molecular Diffusion and Interaction in Various Membrane Environments
Pincet F, Adrien V, Yang R, Delacotte J, Rothman JE, Urbach W, Tareste D. FRAP to Characterize Molecular Diffusion and Interaction in Various Membrane Environments. PLOS ONE 2016, 11: e0158457. PMID: 27387979, PMCID: PMC4936743, DOI: 10.1371/journal.pone.0158457.Peer-Reviewed Original ResearchMeSH KeywordsAnimalsCell MembraneCytoplasmDiffusionFluorescence Recovery After PhotobleachingLipid BilayersLipidsMembranes, ArtificialMiceMicroscopy, ConfocalMunc18 ProteinsRatsReproducibility of ResultsSynapsinsVesicle-Associated Membrane Protein 2ConceptsFluorescence correlation spectroscopySingle-particle trackingCorresponding recovery timeFRAP measurementsDynamics of lipidsDiffusion coefficientCorrelation spectroscopyBrownian motionParticle trackingConfocal microscopeAccurate valuesDiffusive speciesMembrane environmentMolecular diffusionMeasurementsMembrane platformsBehavior of lipidsFRAP experimentsMotion
2013
Preparation and characterization of SNARE-containing nanodiscs and direct study of cargo release through fusion pores
Shi L, Howan K, Shen QT, Wang YJ, Rothman JE, Pincet F. Preparation and characterization of SNARE-containing nanodiscs and direct study of cargo release through fusion pores. Nature Protocols 2013, 8: 935-948. PMID: 23598444, DOI: 10.1038/nprot.2013.048.Peer-Reviewed Original ResearchMeSH KeywordsBiological TransportLiposomesMembrane FusionMicroscopy, FluorescenceModels, BiologicalNanostructuresSynaptosomal-Associated Protein 25Vesicle-Associated Membrane Protein 2ConceptsFusion eventsScaffold proteinFusion poreMembrane scaffold proteinFluorescent lipidFluorescence-based approachFusion machineryAccessible lipidsCognate proteinNanodiscsPore expansionLipid bilayersProteinCargo releaseCargoLipidsPlate readerVAMP2MachineryFluorescenceSnareEncapsulated cargoDirect studyReleaseAssays
2012
SNARE Proteins: One to Fuse and Three to Keep the Nascent Fusion Pore Open
Shi L, Shen QT, Kiel A, Wang J, Wang HW, Melia TJ, Rothman JE, Pincet F. SNARE Proteins: One to Fuse and Three to Keep the Nascent Fusion Pore Open. Science 2012, 335: 1355-1359. PMID: 22422984, PMCID: PMC3736847, DOI: 10.1126/science.1214984.Peer-Reviewed Original ResearchMeSH KeywordsAnimalsCalciumDiffusionLipid BilayersLiposomesMembrane FusionMembrane ProteinsMiceNeurotransmitter AgentsProtein Structure, TertiaryProteolipidsRatsRecombinant Fusion ProteinsSNARE ProteinsSynaptic TransmissionSynaptic VesiclesSynaptosomal-Associated Protein 25Syntaxin 1Vesicle-Associated Membrane Protein 2ConceptsVesicle-associated membrane protein 2Bilayer fusionNative transmembrane domainNascent fusion poresLipid bilayer nanodiscsMembrane protein 2Synchronous neurotransmitter releaseSNARE complexTransmembrane helicesTransmembrane domainBilayer nanodiscsFused bilayersFusion porePore opensFusion partnerBiochemical studiesProtein 2Neurotransmitter releaseNanodiscsSnareEfficient releaseSynaptic transmissionSNAREpinsFusionRelevant time scales
2011
Complexin cross-links prefusion SNAREs into a zigzag array
Kümmel D, Krishnakumar SS, Radoff DT, Li F, Giraudo CG, Pincet F, Rothman JE, Reinisch KM. Complexin cross-links prefusion SNAREs into a zigzag array. Nature Structural & Molecular Biology 2011, 18: 927-933. PMID: 21785414, PMCID: PMC3410656, DOI: 10.1038/nsmb.2101.Peer-Reviewed Original ResearchA conformational switch in complexin is required for synaptotagmin to trigger synaptic fusion
Krishnakumar SS, Radoff DT, Kümmel D, Giraudo CG, Li F, Khandan L, Baguley SW, Coleman J, Reinisch KM, Pincet F, Rothman JE. A conformational switch in complexin is required for synaptotagmin to trigger synaptic fusion. Nature Structural & Molecular Biology 2011, 18: 934-940. PMID: 21785412, PMCID: PMC3668341, DOI: 10.1038/nsmb.2103.Peer-Reviewed Original ResearchMeSH KeywordsAdaptor Proteins, Vesicular TransportAmino Acid SequenceAnimalsBinding SitesCrystallography, X-RayHumansMembrane FusionModels, MolecularMolecular Sequence DataMutagenesis, Site-DirectedNerve Tissue ProteinsProtein Structure, TertiaryRatsSynaptosomal-Associated Protein 25SynaptotagminsSyntaxin 1Vesicle-Associated Membrane Protein 2Complexin activates and clamps SNAREpins by a common mechanism involving an intermediate energetic state
Li F, Pincet F, Perez E, Giraudo CG, Tareste D, Rothman JE. Complexin activates and clamps SNAREpins by a common mechanism involving an intermediate energetic state. Nature Structural & Molecular Biology 2011, 18: 941-946. PMID: 21785413, PMCID: PMC3736826, DOI: 10.1038/nsmb.2102.Peer-Reviewed Original Research