2025
Single-molecule two- and three-colour FRET studies reveal a transition state in SNARE disassembly by NSF
Cheppali S, Li C, Xing W, Sun R, Yang M, Xue Y, Lu S, Yao J, Sun S, Chen C, Sui S. Single-molecule two- and three-colour FRET studies reveal a transition state in SNARE disassembly by NSF. Nature Communications 2025, 16: 250. PMID: 39747074, PMCID: PMC11695992, DOI: 10.1038/s41467-024-55531-0.Peer-Reviewed Original ResearchConceptsN-ethylmaleimide sensitive factorSNARE complexDisassembly of SNARE complexesStable four-helix bundleFour-helix bundleSNARE motifFluorescence spectroscopy approachesMinimal machineryAAA+ ATPasesProtein machineryAdapter proteinVesicle fusionMembrane fusionSyntaxinPhysiological processesSnareProteinN-ethylmaleimideDisassemblySequential disassemblyMachinerySequential pathwayPathwayEukaryotesFusion
2022
Measuring mRNA Decay with Roadblock‐qPCR
Watson MJ, Thoreen CC. Measuring mRNA Decay with Roadblock‐qPCR. Current Protocols 2022, 2: e344. PMID: 35041257, PMCID: PMC8830782, DOI: 10.1002/cpz1.344.Peer-Reviewed Original ResearchConceptsMRNA stabilityPost-transcriptional regulatory stepsPre-existing mRNAsMRNA decay kineticsQuantitative PCRN-ethylmaleimideGene functionGene regulationGeneral transcriptionNascent mRNARegulatory stepEndogenous mRNACDNA poolsGene expressionLiving cellsTranscriptionCell functionMRNAReverse transcriptionCDNA synthesisMRNA levelsCellsTranscriptsRNACommon strategy
1999
Functional interaction of the K-Cl cotransporter (KCC1) with the Na-K-Cl cotransporter in HEK-293 cells
Gillen C, Forbush B. Functional interaction of the K-Cl cotransporter (KCC1) with the Na-K-Cl cotransporter in HEK-293 cells. American Journal Of Physiology 1999, 276: c328-c336. PMID: 9950760, DOI: 10.1152/ajpcell.1999.276.2.c328.Peer-Reviewed Original ResearchConceptsNa-K-Cl cotransporterNa-K-Cl cotransport activityNa-K-ClK-Cl cotransportHEK-293 cellsCotransport activityK-ClHEK-293Activation of Na-K-Cl cotransportCell swellingK-Cl cotransport activityHypotonic cell swellingIntracellular Cl- concentrationFunctional interactionsKCC1External KCotransporterIon-transporting epitheliaCell linesN-ethylmaleimidePhysiological rangeCellsInvestigate functional interactionsCross-talkPretreatmentConstitutive Calcium-independent Release of Toxoplasma gondii Dense Granules Occurs through the NSF/SNAP/SNARE/Rab Machinery*
Chaturvedi S, Qi H, Coleman D, Rodriguez A, Hanson P, Striepen B, Roos D, Joiner K. Constitutive Calcium-independent Release of Toxoplasma gondii Dense Granules Occurs through the NSF/SNAP/SNARE/Rab Machinery*. Journal Of Biological Chemistry 1999, 274: 2424-2431. PMID: 9891012, DOI: 10.1074/jbc.274.4.2424.Peer-Reviewed Original ResearchConceptsN-ethylmaleimide-sensitive fusion proteinDense matrix granulesDense granulesBeta-lactamase secretionMammalian homologueDissociation inhibitorAbsence of ATPPathogenic protozoan parasitesCalcium-independent fashionPermeabilized parasitesFusion proteinProtein componentsProtozoan parasiteCalcium-independent releaseDense granule releaseParasitesMachineryProteinN-ethylmaleimideGuanosine 5'Matrix granulesGranule releaseToxoplasma gondiiGranulesHomologues
1994
Acute Regulation of Na+:HCO3- Cotransport System In Kidney Proximal Tubules
Soleimani M, Bizal G, Hattabaugh Y, Aronson P, Bergman J. Acute Regulation of Na+:HCO3- Cotransport System In Kidney Proximal Tubules. NATO ASI Series 1994, 309-317. DOI: 10.1007/978-3-642-79301-1_36.ChaptersKidney proximal tubulesProximal tubulesKidney proximal tubule cellsProximal tubule cellsCotransport systemFiltered loadBlood pHHCO3- reabsorptionTubule cellsAcute regulatory mechanismsAcute regulationTransport of HCO3N-ethylmaleimideTubulesBasolateral membraneRecent studiesCotransporterLumenTransportersFunctional characteristicsCellsRegulatory mechanismsNumerous typesBloodSecretion
1991
Apical membrane localization of ouabain-sensitive K(+)-activated ATPase activities in rat distal colon
Del Castillo J, Rajendran V, Binder H. Apical membrane localization of ouabain-sensitive K(+)-activated ATPase activities in rat distal colon. American Journal Of Physiology 1991, 261: g1005-g1011. PMID: 1837421, DOI: 10.1152/ajpgi.1991.261.6.g1005.Peer-Reviewed Original ResearchConceptsRat distal colonOuabain-insensitive fractionDistal colonOuabain-sensitive fractionOuabain-insensitive componentHalf-maximal concentrationApical membraneATPase activityColonic mucosaProximal colonColonActivation curveAbsorptive processesAdenosinetriphosphatase activityApical membrane localizationBasolateral membraneHomogenatesN-ethylmaleimideMucosaActivityOmeprazoleChanges in channel properties of acetylcholine receptors during the time course of thiol chemical modifications
Bouzat C, Barrantes F, Sigworth F. Changes in channel properties of acetylcholine receptors during the time course of thiol chemical modifications. Pflügers Archiv - European Journal Of Physiology 1991, 418: 51-61. PMID: 2041725, DOI: 10.1007/bf00370451.Peer-Reviewed Original ResearchConceptsSingle acetylcholine receptor channelsHigh agonist concentrationsPatch-clamp techniqueOpen timeConcentration of NEMSilent periodAcetylcholine receptorsN-ethylmaleimideAcetylcholine receptor channelsAgonist concentrationsChannel open probabilityReceptor channelsAChRMM N-ethylmaleimideTime of exposureRate of occurrenceSingle-channel currentsTime courseNEM treatmentControl receptorsReceptorsSignificant changesTreatmentShort openingsSlight reduction
1990
Apical and basolateral membrane H+ extrusion mechanisms in inner stripe of rabbit outer medullary collecting duct
Hays SR, Alpern RJ. Apical and basolateral membrane H+ extrusion mechanisms in inner stripe of rabbit outer medullary collecting duct. American Journal Of Physiology 1990, 259: f628-f635. PMID: 2171359, DOI: 10.1152/ajprenal.1990.259.4.f628.Peer-Reviewed Original ResearchConceptsUnits/minPHi recoveryInner stripeAcid loadInitial control levelsSCH 28080Luminal Sch-28080ATPase inhibitorBasolateral membraneTriphasic responseExtrusion mechanismSlow alkalinizationRate of alkalinizationControl levelsCell acidificationRabbitsInhibitorsDuctTotal absenceInitial rateAlkalinizationN-ethylmaleimideRecoveryMin
1989
Synaptic vesicles immunoisolated from rat cerebral cortex contain high levels of glutamate
Burger P, Mehl E, Cameron P, Maycox P, Baumert M, Lottspeich F, De Camilli P, Jahn R. Synaptic vesicles immunoisolated from rat cerebral cortex contain high levels of glutamate. Neuron 1989, 3: 715-720. PMID: 2577130, DOI: 10.1016/0896-6273(89)90240-7.Peer-Reviewed Original ResearchConceptsSynaptic vesiclesRat cerebral cortexRat brain synaptic vesiclesProton electrochemical gradientBrain synaptic vesiclesCerebral cortexCytosolic poolMajor excitatory neurotransmitterVesicle membraneElectrochemical gradientAmino acidsVesiclesVesicle hypothesisMammalian CNSExcitatory neurotransmitterGlutamate lossEnrichment of glutamateN-ethylmaleimideStarting fractionGlutamate contentGlutamateHigh levelsL-glutamateCortexExchange experiments
1987
Spectrin Is Associated With Membrane-Bound Actin Filaments in Platelets and Is Hydrolyzed by the Ca2+-Dependent Protease During Platelet Activation
Fox J, Reynolds C, Morrow J, Phillips D. Spectrin Is Associated With Membrane-Bound Actin Filaments in Platelets and Is Hydrolyzed by the Ca2+-Dependent Protease During Platelet Activation. Blood 1987, 69: 537-545. DOI: 10.1182/blood.v69.2.537.537.Peer-Reviewed Original ResearchActin filamentsDependent proteasePlasma membraneRBC spectrinSubmembranous actin filamentsActin-binding proteinsBeta chainMembrane-bound filamentsActin filament networkTriton XImmunoprecipitation experimentsMinor polypeptidesFilament networkSpectrinGlycoprotein IbBrain spectrinDifferential centrifugationPresence of Ca2ProteaseLysatesPolyacrylamide gelsUnstimulated plateletsPolypeptideUnidentified componentsN-ethylmaleimide
1984
The hydrogen ion-pumping adenosine triphosphatase of platelet dense granule membrane. Differences from F1F0- and phosphoenzyme-type ATPases.
Dean G, Fishkes H, Nelson P, Rudnick G. The hydrogen ion-pumping adenosine triphosphatase of platelet dense granule membrane. Differences from F1F0- and phosphoenzyme-type ATPases. Journal Of Biological Chemistry 1984, 259: 9569-9574. PMID: 6204985, DOI: 10.1016/s0021-9258(17)42738-4.Peer-Reviewed Original ResearchConceptsGranule ATPaseGranule membranesMitochondrial membraneATP hydrolysisIon-translocating ATPasesEnergy-transducing membranesDense granule membraneIdentical assay conditionsF1F0-ATPasesSubstrate specificityPlasma membraneMitochondrial ATPaseInhibitor sensitivityATPasesATPaseATPase activityConcentrations of NaN3Cation requirementATPase moleculesMembraneN-ethylmaleimideAssay conditionsAdenosine triphosphataseEfrapeptinOligomycin
1983
Acidification of macrophage and fibroblast endocytic vesicles in vitro.
Galloway C, Dean G, Marsh M, Rudnick G, Mellman I. Acidification of macrophage and fibroblast endocytic vesicles in vitro. Proceedings Of The National Academy Of Sciences Of The United States Of America 1983, 80: 3334-3338. PMID: 6190176, PMCID: PMC394037, DOI: 10.1073/pnas.80.11.3334.Peer-Reviewed Original ResearchConceptsAcidic prelysosomal compartmentEndosome-lysosome fusionATP-dependent acidificationF0-ATPaseEndocytic vesiclesPrelysosomal compartmentAddition of ATPPrelysosomal vacuolesMitochondrial F1LysosomesEndosomesSecretory granulesCultured macrophagesN-ethylmaleimideInternal pHPercoll density gradientsAcidificationPH gradientFibroblastsCompartmentsOrganellesInhibitorsPermeant anionsFluorescein-dextranVacuoles
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