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
2020
Freezing 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
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
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
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