2024
A proteome-wide quantitative platform for nanoscale spatially resolved extraction of membrane proteins into native nanodiscs
Brown C, Ghosh S, McAllister R, Kumar M, Walker G, Sun E, Aman T, Panda A, Kumar S, Li W, Coleman J, Liu Y, Rothman J, Bhattacharyya M, Gupta K. A proteome-wide quantitative platform for nanoscale spatially resolved extraction of membrane proteins into native nanodiscs. Nature Methods 2024, 1-10. PMID: 39609567, DOI: 10.1038/s41592-024-02517-x.Peer-Reviewed Original ResearchTarget membrane proteinsMembrane proteinsMembrane contextSynaptic vesicle membrane proteinVesicle membrane proteinsMammalian membrane proteinsMembrane-active polymersExtraction of membrane proteinsNative nanodiscsOrganellar membranesNative membrane environmentMultiprotein complexesMolecular contextCellular membranesMembrane environmentQuantitative platformBioanalytical approachesExtraction efficiencyOpen-access databasesProteinMembraneExtraction conditionsNanodiscsTarget MPNative top-down analysis of protein complexes from the membrane
Jung W, Panda A, Lee J, Shaw JB, Gupta, K. Native top-down analysis of protein complexes from the membrane. Nature Chemistry (Under Review)Peer-Reviewed Original ResearchStructure and function of the human apoptotic scramblase Xkr4.
Chakraborty S, Feng Z, Lee S, Alvarenga OE, Panda A, Bruni R, Khelashvili G, Gupta K, Accardi A. Structure and function of the human apoptotic scramblase Xkr4. BioRxiv 2024 PMID: 39149361, DOI: 10.1101/2024.08.07.607004.Peer-Reviewed Original ResearchAuthor Correction: Structure of Geobacter cytochrome OmcZ identifies mechanism of nanowire assembly and conductivity
Gu Y, Guberman-Pfeffer M, Srikanth V, Shen C, Giska F, Gupta K, Londer Y, Samatey F, Batista V, Malvankar N. Author Correction: Structure of Geobacter cytochrome OmcZ identifies mechanism of nanowire assembly and conductivity. Nature Microbiology 2024, 9: 2759-2759. PMID: 38684912, DOI: 10.1038/s41564-024-01702-0.Peer-Reviewed Original ResearchA proteome-wide quantitative platform for nanoscale spatially resolved extraction of membrane proteins into native nanodiscs.
Brown C, Ghosh S, McAllister R, Kumar M, Walker G, Sun E, Aman T, Panda A, Kumar S, Li W, Coleman J, Liu Y, Rothman JE, Bhattacharyya M, Gupta K. A proteome-wide quantitative platform for nanoscale spatially resolved extraction of membrane proteins into native nanodiscs. BioRxiv 2024 PMID: 38405833, DOI: 10.1101/2024.02.10.579775.Peer-Reviewed Original Research In PressImaging nanoscale-spatial oligomeric organization of membrane proteins directly from native membranes at single-molecule resolution
Walker G, Brown C, Ge X, Kumar S, Muzumdar M, Gupta K, Bhattacharyya M. Imaging nanoscale-spatial oligomeric organization of membrane proteins directly from native membranes at single-molecule resolution. Biophysical Journal 2024, 123: 348a. DOI: 10.1016/j.bpj.2023.11.2115.Peer-Reviewed Original Research
2023
Isolation and Lipidomic Profiling of Neuronal Lipid Droplets: Unveiling the Lipid Landscape for insights into Neurodegenerative Disorders.
Kumar M, Knapp JA, Gupta K, Ryan TA. Isolation and Lipidomic Profiling of Neuronal Lipid Droplets: Unveiling the Lipid Landscape for insights into Neurodegenerative Disorders. BioRxiv 2023 PMID: 38168251, DOI: 10.1101/2023.12.13.571527.Peer-Reviewed Original ResearchOligomeric organization of membrane proteins from native membranes at nanoscale spatial and single-molecule resolution
Walker G, Brown C, Ge X, Kumar S, Muzumdar M, Gupta K, Bhattacharyya M. Oligomeric organization of membrane proteins from native membranes at nanoscale spatial and single-molecule resolution. Nature Nanotechnology 2023, 19: 85-94. PMID: 38012273, PMCID: PMC10981947, DOI: 10.1038/s41565-023-01547-4.Peer-Reviewed Original ResearchMembrane proteinsNative membranesOligomeric organizationDiverse membrane proteinsMembrane protein biologyNative cell membranesTarget membrane proteinsNative membrane environmentSingle-molecule resolutionSingle-molecule platformReceptor tyrosine kinasesOligomeric distributionNative nanodiscsOligomerization statusProtein biologySmall GTPaseGrowth factor bindingMembrane environmentOligomeric assembliesTyrosine kinaseCritical regulatorOncogenic mutationsCell membraneProteinMembraneRoles for diacylglycerol in synaptic vesicle priming and release revealed by complete reconstitution of core protein machinery
Sundaram R, Chatterjee A, Bera M, Grushin K, Panda A, Li F, Coleman J, Lee S, Ramakrishnan S, Ernst A, Gupta K, Rothman J, Krishnakumar S. Roles for diacylglycerol in synaptic vesicle priming and release revealed by complete reconstitution of core protein machinery. Proceedings Of The National Academy Of Sciences Of The United States Of America 2023, 120: e2309516120. PMID: 37590407, PMCID: PMC10450444, DOI: 10.1073/pnas.2309516120.Peer-Reviewed Original ResearchConceptsCore protein machineryRelease-ready vesiclesSynaptic vesicle primingVesicle primingProtein machinerySingle-molecule imagingSNAREpin assemblyFunctional intermediatesFunctional reconstitutionMunc13DiacylglycerolCoordinated actionMunc18VesiclesMachineryComplete reconstitutionNew roleSelective effectDetailed characterizationChaperonesRate of caReconstitutionVAMP2ComplexinMutationsStudying Membrane Protein–Lipid Specificity through Direct Native Mass Spectrometric Analysis from Tunable Proteoliposomes
Panda A, Brown C, Gupta K. Studying Membrane Protein–Lipid Specificity through Direct Native Mass Spectrometric Analysis from Tunable Proteoliposomes. Journal Of The American Society For Mass Spectrometry 2023, 34: 1917-1927. PMID: 37432128, PMCID: PMC10932607, DOI: 10.1021/jasms.3c00110.Peer-Reviewed Original ResearchConceptsIntegral membrane proteinsMembrane proteinsNative mass spectrometryTrafficking pathwaysPlasma membraneEukaryotic integral membrane proteinsEndoplasmic reticulumBiophysical propertiesMembrane protein assemblySynaptic vesiclesCellular trafficking pathwaysOrganellar membranesLipid specificityTransmembrane proteinProtein assembliesMembrane contextMass spectrometric analysisProteinNative mass spectrometric analysesVAMP2Lipid compositionExogenous ligandsLipid membranesIndividual lipidsMembraneATG9 vesicles comprise the seed membrane of mammalian autophagosomes
Olivas T, Wu Y, Yu S, Luan L, Choi P, Guinn E, Nag S, De Camilli P, Gupta K, Melia T. ATG9 vesicles comprise the seed membrane of mammalian autophagosomes. Journal Of Cell Biology 2023, 222: e202208088. PMID: 37115958, PMCID: PMC10148236, DOI: 10.1083/jcb.202208088.Peer-Reviewed Original ResearchConceptsAtg9 vesiclesMammalian autophagosomesStyrene maleic acid lipid particlesLipid scramblase activityLC3-IIAutophagosomes formAutophagosome membraneMature autophagosomesScramblase activityAutophagosome formationAtg9Lipid transportMembrane growthAutophagosomesNanoscale organizationProtein-mediated transferProteinMembrane surface areaOrganellesVesiclesSeed membraneMembraneLipid particlesLipidsDifferent stagesDirect 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 determinationBilayersTransportersStructure of Geobacter cytochrome OmcZ identifies mechanism of nanowire assembly and conductivity
Gu Y, Guberman-Pfeffer M, Srikanth V, Shen C, Giska F, Gupta K, Londer Y, Samatey F, Batista V, Malvankar N. Structure of Geobacter cytochrome OmcZ identifies mechanism of nanowire assembly and conductivity. Nature Microbiology 2023, 8: 284-298. PMID: 36732469, PMCID: PMC9999484, DOI: 10.1038/s41564-022-01315-5.Peer-Reviewed Original ResearchConceptsNanowire assembliesHigh electron conductivityExtracellular electron transportNanowire networksExtracellular electron acceptorsG. sulfurreducensElectron conductivityCryogenic electron microscopy structureNanowiresHigh conductivityElectron microscopy structureSerine proteasesDifferent biochemical environmentsElectron transportMicroscopy structureDiverse speciesGeobacter speciesSulfurreducensConductivityImportant bacteriaCharge interactionsElectron acceptorBiochemical environmentSpeciesIdentifies mechanisms
2022
Mitoguardin-2–mediated lipid transfer preserves mitochondrial morphology and lipid droplet formation
Hong Z, Adlakha J, Wan N, Guinn E, Giska F, Gupta K, Melia TJ, Reinisch KM. Mitoguardin-2–mediated lipid transfer preserves mitochondrial morphology and lipid droplet formation. Journal Of Cell Biology 2022, 221: e202207022. PMID: 36282247, PMCID: PMC9597353, DOI: 10.1083/jcb.202207022.Peer-Reviewed Original ResearchConceptsEndoplasmic reticulumLipid dropletsProtein-mediated transferLipid transport proteinsLipid droplet formationLD biologyMitochondrial proteinsSecretory pathwayMass spectrometry analysisTerminal domainMitochondrial morphologyTransport proteinsLipid transportersCellular membranesLD metabolismMembrane contactX-ray structureSpectrometry analysisOrganellesGlycerophospholipidsProteinHydrophobic cavityFatty acidsLipidsMembraneDeciphering the molecular organization of Get pathway chaperones through native top-down dissociation of multi-protein complexes
Giska F, Mariappan M, Bhattacharyya M, Gupta K. Deciphering the molecular organization of Get pathway chaperones through native top-down dissociation of multi-protein complexes. Biophysical Journal 2022, 121: 333a. DOI: 10.1016/j.bpj.2021.11.1119.Peer-Reviewed Original Research
2021
Chemoselective restoration of para-azido-phenylalanine at multiple sites in proteins
Arranz-Gibert P, Vanderschuren K, Haimovich A, Halder A, Gupta K, Rinehart J, Isaacs FJ. Chemoselective restoration of para-azido-phenylalanine at multiple sites in proteins. Cell Chemical Biology 2021, 29: 1046-1052.e4. PMID: 34965380, PMCID: PMC10173106, DOI: 10.1016/j.chembiol.2021.12.002.Peer-Reviewed Original ResearchConceptsProtein functionalizationChemical moietiesDiverse chemical moietiesDiverse chemical groupsSite-specific incorporationClick reactionOff-target modificationsDiazotransfer reactionAzide moietyAzide reductionNonstandard amino acidsChemical groupsFunctionalizationMaterials scienceMoietyPAZFFunction of proteinsReactionAzideBroad applicationsPhysiological conditionsParaChemistryAmino acidsKey limitationsA human apolipoprotein L with detergent-like activity kills intracellular pathogens
Gaudet RG, Zhu S, Halder A, Kim BH, Bradfield CJ, Huang S, Xu D, Mamiñska A, Nguyen TN, Lazarou M, Karatekin E, Gupta K, MacMicking JD. A human apolipoprotein L with detergent-like activity kills intracellular pathogens. Science 2021, 373 PMID: 34437126, PMCID: PMC8422858, DOI: 10.1126/science.abf8113.Peer-Reviewed Original ResearchMeSH KeywordsApolipoproteins LBacterial Outer MembraneBacteriolysisCell MembraneCell Membrane PermeabilityCells, CulturedCRISPR-Cas SystemsCytosolDetergentsGene EditingGram-Negative BacteriaGTP-Binding ProteinsHumansImmunity, InnateInterferon-gammaLipoproteinsMicrobial ViabilityO AntigensProtein DomainsSalmonella typhimuriumSolubilityConceptsSingle-particle cryo-electron microscopyCell-autonomous defenseCytosol-invasive bacteriaExpression of hundredsNative mass spectrometryCryo-electron microscopyHuman genesDetergent-like activityHost proteinsLipoprotein nanodiscsMammalian lipidsExtracellular transportImmune cytokine interferonCell typesDetergent-like propertiesApolipoprotein LLife-threatening infectionsPotent bactericidal agentsAnionic membranesProteinCytokine interferonNonimmune cellsMass spectrometryCellsMutagenesis
2018
Identifying key membrane protein lipid interactions using mass spectrometry
Gupta K, Li J, Liko I, Gault J, Bechara C, Wu D, Hopper JTS, Giles K, Benesch JLP, Robinson CV. Identifying key membrane protein lipid interactions using mass spectrometry. Nature Protocols 2018, 13: 1106-1120. PMID: 29700483, PMCID: PMC6049616, DOI: 10.1038/nprot.2018.014.Peer-Reviewed Original Research
2017
Interfacial lipids as modulator of membrane protein oligomerisation
Gupta K, Robinson C. Interfacial lipids as modulator of membrane protein oligomerisation. Acta Crystallographica Section A: Foundations And Advances 2017, 73: c101-c101. DOI: 10.1107/s2053273317094700.Peer-Reviewed Original ResearchInnenrücktitelbild: Native Desorption Electrospray Ionization Liberates Soluble and Membrane Protein Complexes from Surfaces (Angew. Chem. 46/2017)
Ambrose S, Housden N, Gupta K, Fan J, White P, Yen H, Marcoux J, Kleanthous C, Hopper J, Robinson C. Innenrücktitelbild: Native Desorption Electrospray Ionization Liberates Soluble and Membrane Protein Complexes from Surfaces (Angew. Chem. 46/2017). Angewandte Chemie 2017, 129: 14965-14965. DOI: 10.1002/ange.201710562.Peer-Reviewed Original Research