2024
Quantitative single-molecule analysis of assembly and Ca2+-dependent disassembly of synaptotagmin oligomers on lipid bilayers
Li F, Coleman J, Redondo-Morata L, Kalyana Sundaram R, Stroeva E, Rothman J, Pincet F. Quantitative single-molecule analysis of assembly and Ca2+-dependent disassembly of synaptotagmin oligomers on lipid bilayers. Communications Biology 2024, 7: 1608. PMID: 39627539, PMCID: PMC11615320, DOI: 10.1038/s42003-024-07317-9.Peer-Reviewed Original ResearchConceptsSyt-1Lipid bilayerRing-like oligomersCa2+-evoked releaseSynaptotagmin-1Single-molecule imaging methodsSynaptic vesiclesBiochemical evidencePhysiological requirementsOligomerizationAnalysis of assembliesBilayerOligomersCa2+LipidAssemblyCa2Classes of oligomersMutationsVesiclesDisassemblyEvoked releasePhotosensitive Nanoprobes for Rapid High Purity Isolation and Size‐Specific Enrichment of Synthetic and Extracellular Vesicle Subpopulations (Adv. Funct. Mater. 34/2024)
Weerakkody J, Tseng T, Topper M, Thoduvayil S, Radhakrishnan A, Pincet F, Kyriakides T, Gunasekara R, Ramakrishnan S. Photosensitive Nanoprobes for Rapid High Purity Isolation and Size‐Specific Enrichment of Synthetic and Extracellular Vesicle Subpopulations (Adv. Funct. Mater. 34/2024). Advanced Functional Materials 2024, 34 DOI: 10.1002/adfm.202470191.Peer-Reviewed Original ResearchVesicle biogenesisExtracellular vesicle subpopulationsLipid nanoprobesVesicle subpopulationsNative extracellular vesiclesVesicle populationsSeparate vesiclesPurity isolationExtracellular vesiclesVesiclesCellular originHydrophobic interactionsBiogenesisSize variabilitySubpopulationsIsolatesExtracellularDual-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 ResearchPhotosensitive Nanoprobes for Rapid High Purity Isolation and Size‐Specific Enrichment of Synthetic and Extracellular Vesicle Subpopulations
Weerakkody J, Tseng T, Topper M, Thoduvayil S, Radhakrishnan A, Pincet F, Kyriakides T, Gunasekara R, Ramakrishnan S. Photosensitive Nanoprobes for Rapid High Purity Isolation and Size‐Specific Enrichment of Synthetic and Extracellular Vesicle Subpopulations. Advanced Functional Materials 2024, 34 PMID: 39372670, PMCID: PMC11452007, DOI: 10.1002/adfm.202400390.Peer-Reviewed Original ResearchExtracellular vesicle subpopulationsVesicle subpopulationsIsolation of vesiclesPurity extracellular vesiclesRelease of vesiclesAnalysis of nucleic acidsNear-native formLarge-scale isolationLipid nanoprobesDownstream analysisPurity isolationEfficient isolationVesiclesSynthetic vesiclesNucleic acidsExtracellular vesiclesIsolation methodIsolatesBiomarker discoveryExposure to lightSubpopulationsEnrichmentProteinComplex biological mediaCleavage
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 actionProteinAssemblyChaperonesSynaptotagminExocytosisBilayersTurbocharging synaptic transmission
Rothman J, Grushin K, Bera M, Pincet F. Turbocharging synaptic transmission. FEBS Letters 2023, 597: 2233-2249. PMID: 37643878, DOI: 10.1002/1873-3468.14718.Peer-Reviewed Original Research
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
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 compartmentVesicle Tubulation with Self‐Assembling DNA Nanosprings
Grome M, Zhang Z, Pincet F, Lin C. Vesicle Tubulation with Self‐Assembling DNA Nanosprings. Angewandte Chemie 2018, 130: 5428-5432. DOI: 10.1002/ange.201800141.Peer-Reviewed Original ResearchMembrane-deforming proteinsDNA origami designMembrane tubulationMembrane tubulesMembrane curvatureVesicle tubulationMembrane surface coverageDNA structureLipid bilayersTubulationNanospringsTube morphologyIntricate interplayArtificial nanomachinesVesicle deformationSpherical vesiclesNanotechnologyMajor goalProteinDNAVesiclesNanomachinesBioengineeringDetergentsMorphology
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 ResearchConceptsMembrane 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
2015
Accelerating SNARE‐Mediated Membrane Fusion by DNA–Lipid Tethers
Xu W, Wang J, Rothman J, Pincet F. Accelerating SNARE‐Mediated Membrane Fusion by DNA–Lipid Tethers. Angewandte Chemie 2015, 127: 14596-14600. DOI: 10.1002/ange.201506844.Peer-Reviewed Original ResearchDNA-lipidMembrane-distal endMembrane-proximal endArtificial tetherSNARE functionCore machinerySNARE proteinsProtein functionTarget membraneMembrane fusionBiological processesNative proteinBase pairsLipid mixingMaximum fusion rateProgrammable toolsBase-pair hybridizationProteinSnareMembraneFusionMachineryTetherNucleotidesVesiclesFormation of Giant Unilamellar Proteo-Liposomes by Osmotic Shock
Motta I, Gohlke A, Adrien V, Li F, Gardavot H, Rothman JE, Pincet F. Formation of Giant Unilamellar Proteo-Liposomes by Osmotic Shock. Langmuir 2015, 31: 7091-7099. PMID: 26038815, PMCID: PMC4950989, DOI: 10.1021/acs.langmuir.5b01173.Peer-Reviewed Original ResearchConceptsGiant unilamellar vesiclesLipid-anchored proteinsOsmotic shockTrans-membrane proteinsSingle giant unilamellar vesiclesProtein substratesPeripheral proteinsSpecific lipidsDifferent proteinsPhotobleaching experimentsFluorescence recoveryCell membraneProteinLarge vesiclesPhysiological conditionsModel systemUnilamellar vesiclesPhospholipid bilayersVesiclesSimple generic methodPrevious dataMembraneHigh concentrationsLipidsBilayers
2005
Indirect 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 medium
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