2024
Dual-Ring SNAREpin Machinery Tuning for Fast Synaptic Vesicle Fusion
Caruel M, Pincet F. Dual-Ring SNAREpin Machinery Tuning for Fast Synaptic Vesicle Fusion. Biomolecules 2024, 14: 600. PMID: 38786007, PMCID: PMC11117985, DOI: 10.3390/biom14050600.Peer-Reviewed Original Research
2023
Synaptophysin chaperones the assembly of 12 SNAREpins under each ready-release vesicle
Bera M, Radhakrishnan A, Coleman J, Sundaram R, Ramakrishnan S, Pincet F, Rothman J. Synaptophysin chaperones the assembly of 12 SNAREpins under each ready-release vesicle. Proceedings Of The National Academy Of Sciences Of The United States Of America 2023, 120: e2311484120. PMID: 37903271, PMCID: PMC10636311, DOI: 10.1073/pnas.2311484120.Peer-Reviewed Original ResearchConceptsSpecific molecular functionsSynaptic vesicle protein synaptophysinTarget membrane bilayerSensor synaptotagminSNARE proteinsMolecular functionsMembrane proteinsSNAREpinsReceptor vesiclesSingle-molecule measurementsGene knockoutMembrane bilayerLipid bilayersProtein synaptophysinVesiclesDetergent extractsHexamer structureSYPMechanism of actionProteinAssemblyChaperonesSynaptotagminExocytosisBilayers
2022
The beginning and the end of SNARE‐induced membrane fusion
Mion D, Bunel L, Heo P, Pincet F. The beginning and the end of SNARE‐induced membrane fusion. FEBS Open Bio 2022, 12: 1958-1979. PMID: 35622519, PMCID: PMC9623537, DOI: 10.1002/2211-5463.13447.Peer-Reviewed Original Research
2019
SNARE machinery is optimized for ultrafast fusion
Manca F, Pincet F, Truskinovsky L, Rothman JE, Foret L, Caruel M. SNARE machinery is optimized for ultrafast fusion. Proceedings Of The National Academy Of Sciences Of The United States Of America 2019, 116: 2435-2442. PMID: 30700546, PMCID: PMC6377469, DOI: 10.1073/pnas.1820394116.Peer-Reviewed Original Research
2018
High-Throughput Monitoring of Single Vesicle Fusion Using Freestanding Membranes and Automated Analysis
Ramakrishnan S, Gohlke A, Li F, Coleman J, Xu W, Rothman JE, Pincet F. High-Throughput Monitoring of Single Vesicle Fusion Using Freestanding Membranes and Automated Analysis. Langmuir 2018, 34: 5849-5859. PMID: 29694054, DOI: 10.1021/acs.langmuir.8b00116.Peer-Reviewed Original ResearchConceptsMembrane fusionFusion eventsSoluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) proteinsSNARE-dependent membrane fusionIndividual vesicle fusion eventsFactor attachment protein receptor proteinsN-ethylmaleimide-sensitive factor attachment protein receptor proteinsT-SNARE proteinsSingle-vesicle fusionProtein receptor proteinsVesicle fusion eventsMobility of proteinsVesicle dockingContent releaseVesicle fusionHigh-throughput monitoringPlanar membranesReceptor proteinLipid mixingProteinLipid bilayersVesiclesCorrect reconstitutionMembraneAqueous compartment
2017
Actual fusion efficiency in the lipid mixing assay - Comparison between nanodiscs and liposomes
François-Martin C, Pincet F. Actual fusion efficiency in the lipid mixing assay - Comparison between nanodiscs and liposomes. Scientific Reports 2017, 7: 43860. PMID: 28266607, PMCID: PMC5339690, DOI: 10.1038/srep43860.Peer-Reviewed Original Research
2016
A Programmable DNA Origami Platform to Organize SNAREs for Membrane Fusion
Xu W, Nathwani B, Lin C, Wang J, Karatekin E, Pincet F, Shih W, Rothman JE. A Programmable DNA Origami Platform to Organize SNAREs for Membrane Fusion. Journal Of The American Chemical Society 2016, 138: 4439-4447. PMID: 26938705, PMCID: PMC4950518, DOI: 10.1021/jacs.5b13107.Peer-Reviewed Original ResearchMeSH KeywordsDNADNA, Single-StrandedLipid BilayersLiposomesMembrane FusionProtein BindingSNARE ProteinsVesicular Transport ProteinsConceptsMembrane fusionSoluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) complexCore molecular machineryMembrane fusion eventsProtein receptor complexMembrane fusion processMolecular machineryDNA origami platformTarget membraneAuxiliary proteinsIntracellular communicationDocking stepSingle-event levelReceptor complexLipid mixingSmall unilamellar vesiclesLipid bilayersSnareFundamental processesVesiclesUnilamellar vesiclesTraffickingMachineryProteinFusion
2014
A Half-Zippered SNARE Complex Represents a Functional Intermediate in Membrane Fusion
Li F, Kümmel D, Coleman J, Reinisch KM, Rothman JE, Pincet F. A Half-Zippered SNARE Complex Represents a Functional Intermediate in Membrane Fusion. Journal Of The American Chemical Society 2014, 136: 3456-3464. PMID: 24533674, PMCID: PMC3985920, DOI: 10.1021/ja410690m.Peer-Reviewed Original ResearchMeSH KeywordsAnimalsCell MembraneKineticsMembrane FusionMiceModels, MolecularProtein Structure, TertiaryRatsSNARE ProteinsThermodynamicsConceptsN-terminal domainMembrane fusionV-SNARET-SNAREsRecent biophysical studiesC-terminal portionSNARE complexTransmembrane domainRegulatory proteinsFunctional intermediatesC-terminusDistinct functionsN-terminusMolecular mechanismsConformational rearrangementsBiophysical studiesVital regulatorZippering mechanismRate-limiting stepBiological membranesSnareFusionComplexinMultiple stagesZippering
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 ResearchConceptsFusion 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
A 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 ResearchComplexin 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 ResearchCD9 tetraspanin generates fusion competent sites on the egg membrane for mammalian fertilization
Jégou A, Ziyyat A, Barraud-Lange V, Perez E, Wolf J, Pincet F, Gourier C. CD9 tetraspanin generates fusion competent sites on the egg membrane for mammalian fertilization. Proceedings Of The National Academy Of Sciences Of The United States Of America 2011, 108: 10946-10951. PMID: 21690351, PMCID: PMC3131345, DOI: 10.1073/pnas.1017400108.Peer-Reviewed Original ResearchConceptsAdhesion sitesFusion-competent sitesWild-type eggsCD9 tetraspaninEgg membrane proteinsSperm-egg fusionMembrane proteinsGamete membranesMammalian fertilizationCompetent sitesGamete interactionAdhesion eventsTight proximityEgg membraneCD9TetraspaninsEggsFertilizationMembraneStrong-interaction modesAdhesion probabilitySitesFusionInteraction modesProtein
2009
Quantification of phase transitions of lipid mixtures from bilayer to non-bilayer structures: Model, experimental validation and implication on membrane fusion
Xu W, Pincet F. Quantification of phase transitions of lipid mixtures from bilayer to non-bilayer structures: Model, experimental validation and implication on membrane fusion. Chemistry And Physics Of Lipids 2009, 163: 280-285. PMID: 20018184, DOI: 10.1016/j.chemphyslip.2009.12.002.Peer-Reviewed Original ResearchConceptsPhase transition
2007
Energetics and dynamics of SNAREpin folding across lipid bilayers
Li F, Pincet F, Perez E, Eng WS, Melia TJ, Rothman JE, Tareste D. Energetics and dynamics of SNAREpin folding across lipid bilayers. Nature Structural & Molecular Biology 2007, 14: 890-896. PMID: 17906638, DOI: 10.1038/nsmb1310.Peer-Reviewed Original ResearchMeSH KeywordsAnimalsLipid BilayersMembrane FusionMiceProtein ConformationProtein FoldingRatsSNARE ProteinsSurface Properties
2005
Spontaneous and Reversible Switch from Amphiphilic to Oil-Like Structures
Pincet F, Tareste D, Amar M, Perez É. Spontaneous and Reversible Switch from Amphiphilic to Oil-Like Structures. Physical Review Letters 2005, 95: 218101. PMID: 16384186, DOI: 10.1103/physrevlett.95.218101.Peer-Reviewed Original ResearchIndirect evidence of submicroscopic pores in giant unilamelar vesicles
Rodriguez N, Heuvingh J, Pincet F, Cribier S. Indirect evidence of submicroscopic pores in giant unilamelar vesicles. Biochimica Et Biophysica Acta 2005, 1724: 281-287. PMID: 15978732, DOI: 10.1016/j.bbagen.2005.04.028.Peer-Reviewed Original ResearchConceptsPore-like structuresGiant unilamellar vesiclesOuter leafletCell membraneTransient poresFluorescent lipidLipid distributionCell compartmentVesiclesGiant vesiclesModel systemUnilamellar vesiclesLipid redistributionGiant unilamelar vesiclesMembrane defectsLipidsMembraneFormation of poresMembrane ruptureSubmicroscopic poresLeafletsExchange of matterHemifusionFluorescent labelsOuter mediumJohnson-Kendall-Roberts Theory Applied to Living Cells
Chu YS, Dufour S, Thiery JP, Perez E, Pincet F. Johnson-Kendall-Roberts Theory Applied to Living Cells. Physical Review Letters 2005, 94: 028102. PMID: 15698233, DOI: 10.1103/physrevlett.94.028102.Peer-Reviewed Original Research
2004
Hemifusion and fusion of giant vesicles induced by reduction of inter-membrane distance
Heuvingh J, Pincet F, Cribier S. Hemifusion and fusion of giant vesicles induced by reduction of inter-membrane distance. The European Physical Journal E 2004, 14: 269-276. PMID: 15338438, DOI: 10.1140/epje/i2003-10151-2.Peer-Reviewed Original ResearchConceptsInter-membrane distanceGiant unilamellar vesiclesInner leafletMembrane fusionOuter leafletMeans of micromanipulationCommon functionFluorescence microscopyProbe redistributionVesiclesInfluenza virus hemagglutininGiant vesiclesDNA basesModel systemHemifusionUnilamellar vesiclesActual fusionVirus hemagglutininMembraneContact zoneFusionHead groupsGood agreementProteinLeaflets