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 actionProteinAssemblyChaperonesSynaptotagminExocytosisBilayersDirect determination of oligomeric organization of integral membrane proteins and lipids from intact customizable bilayer
Panda A, Giska F, Duncan A, Welch A, Brown C, McAllister R, Hariharan P, Goder J, Coleman J, Ramakrishnan S, Pincet F, Guan L, Krishnakumar S, Rothman J, Gupta K. Direct determination of oligomeric organization of integral membrane proteins and lipids from intact customizable bilayer. Nature Methods 2023, 20: 891-897. PMID: 37106230, PMCID: PMC10932606, DOI: 10.1038/s41592-023-01864-5.Peer-Reviewed Original ResearchConceptsIntegral membrane proteinsMembrane proteinsOligomeric organizationOligomeric stateNative mass spectrometry analysisFunctional oligomeric stateKey membrane componentMass spectrometry analysisNMS analysisTarget membraneLipid bindingMembrane componentsProteolipid vesiclesMembrane compositionLipid compositionSpectrometry analysisLipid membranesNeurotransmitter releaseProteinMembraneLipidsMembrane propertiesDirect determinationBilayersTransporters
2020
In vitro physiological membrane‐on‐a‐chip and its application in cell and neuronal biology
Heo P, Rothman J, Pincet F. In vitro physiological membrane‐on‐a‐chip and its application in cell and neuronal biology. The FASEB Journal 2020, 34: 1-1. DOI: 10.1096/fasebj.2020.34.s1.08637.Peer-Reviewed Original ResearchNeuronal biologyTail-anchored proteinsPost-translational insertionMost biological processesAreas of biologyVesicle traffickingProtein insertionER membranePhysiological lipid compositionBiological processesMolecular mechanismsBiologyLipid compositionPhysiological membranesModel membranesMembraneInvaluable insightsCellsTraffickingProteinInsertionPatch-clamp amplifierΑ-synucleinopathiesCompositionBilayers
2019
Highly Reproducible Physiological Asymmetric Membrane with Freely Diffusing Embedded Proteins in a 3D‐Printed Microfluidic Setup
Heo P, Ramakrishnan S, Coleman J, Rothman JE, Fleury J, Pincet F. Highly Reproducible Physiological Asymmetric Membrane with Freely Diffusing Embedded Proteins in a 3D‐Printed Microfluidic Setup. Small 2019, 15: e1900725. PMID: 30977975, DOI: 10.1002/smll.201900725.Peer-Reviewed Original ResearchConceptsMost biological processesLipid leafletAreas of biologyEmbedded proteinsBiological processesRelevant lipidsProteinAsymmetric bilayersPhysiological conditionsModel membranesPlanar bilayersBilayer formation processInvaluable insightsBilayersConfocal microscopeMembraneLipidsTransmembraneBiologyLeafletsMicrofluidic setupRecapitulation
2015
Formation 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
2008
The Surface Force Apparatus to Reveal the Energetics of Biomolecules Assembly. Application to DNA Bases Pairing and SNARE Fusion Proteins Folding
Perez E, Li F, Tareste D, Pincet F. The Surface Force Apparatus to Reveal the Energetics of Biomolecules Assembly. Application to DNA Bases Pairing and SNARE Fusion Proteins Folding. Cellular And Molecular Bioengineering 2008, 1: 240. DOI: 10.1007/s12195-008-0025-7.Peer-Reviewed Original ResearchSNARE proteinsLipid bilayersMolecular binding energyIntracellular fusion eventsSurface force apparatusIntermediate assembly statesSNAREpin assemblyNanoscale resolutionBiomolecule assemblyForce apparatusSFA measurementsBiological moleculesBinding energiesLipid headgroupsFusion eventsMembrane fusionFusion proteinInteraction energyConformational changesAssembly stateAdhesion forceDNA basesProteinBilayersEnergy
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 Research
2004
Can Small Complex Chains Be Treated as Polymers?
Gourier C, Pincet F, Le Bouar T, Zhang Y, Esnault J, Mallet J, Sinay P, Perez E. Can Small Complex Chains Be Treated as Polymers? Macromolecules 2004, 37: 8778-8784. DOI: 10.1021/ma035905m.Peer-Reviewed Original Research
1999
Bilayers of neutral lipids bear a small but significant charge
Pincet F, Cribier S, Perez E. Bilayers of neutral lipids bear a small but significant charge. The European Physical Journal B 1999, 11: 127-130. DOI: 10.1007/bf03219162.Peer-Reviewed Original ResearchNeutral lipid bilayersLipid bilayersSurface force measurementsPure waterPhosphatidyl choline bilayerAqueous mediaElectrophoretic measurementsHigh salt concentrationsSurface forcesNegative chargeSignificant chargeWeak forcesAdhesion energySalt concentrationSalt solutionRepulsionBilayersLess chargeWeak repulsionSaltChargeForce measurementsSmall amountWater
1997
Quantitative Analysis of Holes in Supported Bilayers Providing the Adsorption Energy of Surfactants on Solid Substrate
Bassereau P, Pincet F. Quantitative Analysis of Holes in Supported Bilayers Providing the Adsorption Energy of Surfactants on Solid Substrate. Langmuir 1997, 13: 7003-7007. DOI: 10.1021/la970515c.Peer-Reviewed Original ResearchSecond monolayerFirst monolayerMode atomic force microscopy experimentsAtomic force microscopy experimentsForce microscopy experimentsMicroscopy experimentsHolesSupported bilayersDesorption of lipidRange 1Ratio measurementsAdsorption energyMonolayersEnergySolid surfaceBilayersSolid substrateMixed bilayersLow adsorption energyAmphiphilic moleculesQuantitative analysisMeasurementsSimple techniqueNew simple techniqueDOPC