2024
Kinetic study of membrane protein interactions: from three to two dimensions
Adrien V, Reffay M, Taulier N, Verchère A, Monlezun L, Picard M, Ducruix A, Broutin I, Pincet F, Urbach W. Kinetic study of membrane protein interactions: from three to two dimensions. Scientific Reports 2024, 14: 882. PMID: 38195620, PMCID: PMC10776792, DOI: 10.1038/s41598-023-50827-5.Peer-Reviewed Original ResearchConceptsMembrane proteinsMembrane protein interactionsProtein-protein interactionsProtein complexesProtein interactionsMembrane environmentOpposite membranesBacterial efflux pumpsProtein behaviorProtein systemsMolecular interactionsEfflux pumpsProteinExploration distanceMembraneFluorescence recovery experimentsInteractionBinding rateBinding constantsComplexes
2023
Direct 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
2022
The Get1/2 insertase forms a channel to mediate the insertion of tail-anchored proteins into the ER
Heo P, Culver J, Miao J, Pincet F, Mariappan M. The Get1/2 insertase forms a channel to mediate the insertion of tail-anchored proteins into the ER. Cell Reports 2022, 42: 111921. PMID: 36640319, PMCID: PMC9932932, DOI: 10.1016/j.celrep.2022.111921.Peer-Reviewed Original ResearchConceptsTransmembrane domainTA proteinsSingle C-terminal transmembrane domainC-terminal transmembrane domainTail-anchored (TA) proteinsTail-anchored proteinsEndoplasmic reticulum membraneGet3Reticulum membraneChannel functionInsertaseBulk fluorescenceAqueous channelsProteinChannel activityMutation analysisMembraneMicrofluidic assayTranslocaseYeastComplexesInsertionTranslocationHydrophilic segmentsBindingActuating tension-loaded DNA clamps drives membrane tubulation
Liu L, Xiong Q, Xie C, Pincet F, Lin C. Actuating tension-loaded DNA clamps drives membrane tubulation. Science Advances 2022, 8: eadd1830. PMID: 36223466, PMCID: PMC9555772, DOI: 10.1126/sciadv.add1830.Peer-Reviewed Original ResearchConceptsDNA clampMembrane tubulationMembrane dynamicsMembrane-remodeling eventsVesicle tubulationConformational changesSpatiotemporal controlDNA signalsCell membraneDNA nanostructuresTubulationMembrane deformationClosed stateOpen stateSelf-assembled DNA nanostructuresOrganismsProteinMembrane tubeArtificial systemsTube widthMembraneDynamicsThe 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
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-synucleinopathiesCompositionBilayersFreezing and piercing of in vitro asymmetric plasma membrane by α-synuclein
Heo P, Pincet F. Freezing and piercing of in vitro asymmetric plasma membrane by α-synuclein. Communications Biology 2020, 3: 148. PMID: 32235856, PMCID: PMC7109109, DOI: 10.1038/s42003-020-0883-7.Peer-Reviewed Original ResearchMeSH KeywordsAlpha-SynucleinCell MembraneElectric CapacitanceFluorescence Recovery After PhotobleachingHydrophobic and Hydrophilic InteractionsLab-On-A-Chip DevicesMembrane FluidityMembrane LipidsMembrane PotentialsMembranes, ArtificialMicrofluidic Analytical TechniquesNeuronsProtein AggregatesProtein Aggregation, PathologicalProtein BindingProtein ConformationStructure-Activity RelationshipSynucleinopathiesConceptsPlasma membraneMembrane-bound proteinsAccumulation of aggregatesΑ-synucleinCytosolic leafletMembrane topologyMembrane hydrophobic coreCytosolic proteinsProteinExtracellular onesHydrophobic corePathological roleDiscrete sizesMembraneLeafletsMembrane capacitanceNeurological diseasesLipidsAccumulationMicrofluidic setup
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
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 compartmentRearrangements under confinement lead to increased binding energy of Synaptotagmin‐1 with anionic membranes in Mg2+ and Ca2+
Gruget C, Coleman J, Bello O, Krishnakumar SS, Perez E, Rothman JE, Pincet F, Donaldson SH. Rearrangements under confinement lead to increased binding energy of Synaptotagmin‐1 with anionic membranes in Mg2+ and Ca2+. FEBS Letters 2018, 592: 1497-1506. PMID: 29578584, DOI: 10.1002/1873-3468.13040.Peer-Reviewed Original Research
2016
Endothelial basement membrane laminin 511 is essential for shear stress response
Di Russo J, Luik A, Yousif L, Budny S, Oberleithner H, Hofschröer V, Klingauf J, van Bavel E, Bakker E, Hellstrand P, Bhattachariya A, Albinsson S, Pincet F, Hallmann R, Sorokin L. Endothelial basement membrane laminin 511 is essential for shear stress response. The EMBO Journal 2016, 36: 183-201. PMID: 27940654, PMCID: PMC5239996, DOI: 10.15252/embj.201694756.Peer-Reviewed Original ResearchConceptsCell-cell adhesion strengthLaminin-511Junctional complexesVE-cadherinIntegrin-mediated interactionsEndothelial basement membrane componentVinculin-positive focal adhesionsCatenin associationFocal adhesionsStress responseCell junctionsMembrane componentsBasement membrane componentsBasement membraneArterial endotheliumCortical stiffnessMechanotransductionPECAM-1Firm anchorageEndothelial basement membraneJunctional associationMembraneConsiderable informationComplexesAdhesion
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
2014
CX3CL1, a chemokine finely tuned to adhesion: critical roles of the stalk glycosylation and the membrane domain
Ostuni M, Guellec J, Hermand P, Durand P, Combadière C, Pincet F, Deterre P. CX3CL1, a chemokine finely tuned to adhesion: critical roles of the stalk glycosylation and the membrane domain. Biology Open 2014, 3: 1173-1182. PMID: 25395671, PMCID: PMC4265755, DOI: 10.1242/bio.20149845.Peer-Reviewed Original ResearchCytosolic domainTransmembrane domainChemokine domainMucin stalkMembrane domainsHigh glycosylationFunctional rolePatrolling behaviorFunctional adhesion assaysAdhesion assaysCritical roleGlycosylationDomainCytoskeletonPermanent aggregationStructural analysisStalkAdhesionRoleMembraneCX3CL1CellsAssaysReceptors
2013
COPI buds 60-nm lipid droplets from reconstituted water–phospholipid–triacylglyceride interfaces, suggesting a tension clamp function
Thiam AR, Antonny B, Wang J, Delacotte J, Wilfling F, Walther TC, Beck R, Rothman JE, Pincet F. COPI buds 60-nm lipid droplets from reconstituted water–phospholipid–triacylglyceride interfaces, suggesting a tension clamp function. Proceedings Of The National Academy Of Sciences Of The United States Of America 2013, 110: 13244-13249. PMID: 23901109, PMCID: PMC3746913, DOI: 10.1073/pnas.1307685110.Peer-Reviewed Original ResearchMeSH KeywordsADP-Ribosylation Factor 1AnimalsCoat Protein Complex IEscherichia coliHumansHydrophobic and Hydrophilic InteractionsLipid BilayersMicroscopy, ElectronPhosphatidylcholinesPhosphatidylethanolaminesPhospholipidsProtein TransportSf9 CellsSpectrometry, FluorescenceSpodopteraSurface TensionTransport VesiclesTriglyceridesWaterConceptsCoat protein complex ILipid dropletsCoat protein complexTransport vesicle formationProtein complex ILipid droplet targetingBud vesiclesCOPI subunitsProtein complexesCoat componentsNeighboring proteinsIntracellular traffickingVesicle formationComplex IClamp functionPossible direct interactionDirect interactionPhospholipid packingUnsuitable targetsProteinBilayer membranesCoatomerMembraneOrganellesProtomers
2011
CD9 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
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
2001
Short-range specific forces are able to induce hemifusion
Pincet F, Lebeau L, Cribier S. Short-range specific forces are able to induce hemifusion. European Biophysics Journal 2001, 30: 91-97. PMID: 11409468, DOI: 10.1007/s002490100131.Peer-Reviewed Original ResearchConceptsDepletion forcesInterbilayer distanceAdditional attractive forceAmphiphilic moleculesNucleic basesPolar headBiological membranesLipidic systemsModel membranesPossible structuresFluorescent lipid analogsAttractive forceLow water contentLamellar structureLipid analoguesExtra attractionLipid rearrangementBase pairingWater contentMembrane behaviorNucleosidesPhase transitionMembraneMoleculesAdherent vesicles