2023
Roles 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 caReconstitutionVAMP2ComplexinMutations
2022
Munc13 structural transitions and oligomers that may choreograph successive stages in vesicle priming for neurotransmitter release
Grushin K, Kalyana Sundaram RV, Sindelar CV, Rothman JE. Munc13 structural transitions and oligomers that may choreograph successive stages in vesicle priming for neurotransmitter release. Proceedings Of The National Academy Of Sciences Of The United States Of America 2022, 119: e2121259119. PMID: 35135883, PMCID: PMC8851502, DOI: 10.1073/pnas.2121259119.Peer-Reviewed Original Research
2015
Dimeric Organization of Blood Coagulation Factor VIII bound to Lipid Nanotubes
Dalm D, Galaz-Montoya J, Miller J, Grushin K, Villalobos A, Koyfman A, Schmid M, Stoilova-McPhie S. Dimeric Organization of Blood Coagulation Factor VIII bound to Lipid Nanotubes. Scientific Reports 2015, 5: 11212. PMID: 26082135, PMCID: PMC4469981, DOI: 10.1038/srep11212.Peer-Reviewed Original ResearchConceptsMembrane-bound factor VIIIMembrane-binding domainCryo-electron microscopyMembrane-bound structuresSerine protease factor IXaDeficiency of FVIIIDimeric organizationSubtomogram averagingComplex assemblyHelical reconstructionLipid nanotubesCritical functionsFactor IXaBlood coagulationBlood coagulation factor VIIIStructural informationCrystal structureCritical stepPlatelet membranesPlatelet surfaceSevere bleeding disorderCoagulation factor VIIIIntermediate resolutionBleeding disorderMembrane
2014
Lipid nanotechnologies for structural studies of membrane‐associated proteins
Stoilova‐McPhie S, Grushin K, Dalm D, Miller J. Lipid nanotechnologies for structural studies of membrane‐associated proteins. Proteins Structure Function And Bioinformatics 2014, 82: 2902-2909. PMID: 24957666, PMCID: PMC5292012, DOI: 10.1002/prot.24631.Peer-Reviewed Original ResearchConceptsMembrane-associated proteinsLipid nanotechnologyLipid nanotubesAqueous solutionStructural studiesStructure determinationCryo-EMND technologiesCryo-electron microscopyLipid nanodisksScaffold proteinMembrane curvatureMacromolecular organizationRich membranesLipid bilayersPhysiological environmentProteinNanotechnologyLipid compositionNanotubesPhysiological conditionsAtomic structureFunctional structureProof of principleNanodisksHelical organization of blood coagulation factor VIII on lipid nanotubes.
Miller J, Dalm D, Koyfman A, Grushin K, Stoilova-McPhie S. Helical organization of blood coagulation factor VIII on lipid nanotubes. Journal Of Visualized Experiments 2014 PMID: 24961276, PMCID: PMC4126079, DOI: 10.3791/51254.Peer-Reviewed Original ResearchConceptsCryo-electron microscopyFVIII formsThree-dimensional structure analysisMembrane-bound stateMembrane-bound structuresSerine protease factor IXaBlood coagulation factor VIIIHelical organizationDeficiency of FVIIISequence proteinFunctional complexLipid nanotubesCryo-EMDetailed protocolProteolytic activationLipid nanotechnologyStructure analysisCoagulation factor VIIIFVIII bindsPowerful approachProteinFunctional structureStructural informationPlatelet membranesMembraneLack of recombinant factor VIII B‐domain induces phospholipid vesicle aggregation: implications for the immunogenicity of factor VIII
Grushin K, Miller J, Dalm D, Parker E, Healey J, Lollar P, Stoilova-McPhie S. Lack of recombinant factor VIII B‐domain induces phospholipid vesicle aggregation: implications for the immunogenicity of factor VIII. Haemophilia 2014, 20: 723-731. PMID: 24750465, PMCID: PMC4149818, DOI: 10.1111/hae.12421.Peer-Reviewed Original ResearchConceptsFVIII formsB domainFVIII-BDDPhospholipid vesicle aggregationFactor VIII B domainCryo-electron microscopyMembrane-bound statePhospholipid vesiclesMembrane-bound formSerine protease factor IXaPhysiological conditionsFVIII B domainSecondary structure distributionCryo-EMVesicle aggregationBiophysical propertiesCircular dichroismProtein therapeuticsVesiclesTenase complexPhospholipid membranesPlatelet surfaceHuman factor VIIIFactor IXaHereditary bleeding disorders