2014
Molecular Motions Involved in Na-K-Cl Cotransporter-mediated Ion Transport and Transporter Activation Revealed by Internal Cross-linking between Transmembrane Domains 10 and 11/12*
Monette MY, Somasekharan S, Forbush B. Molecular Motions Involved in Na-K-Cl Cotransporter-mediated Ion Transport and Transporter Activation Revealed by Internal Cross-linking between Transmembrane Domains 10 and 11/12*. Journal Of Biological Chemistry 2014, 289: 7569-7579. PMID: 24451383, PMCID: PMC3953270, DOI: 10.1074/jbc.m113.542258.Peer-Reviewed Original ResearchMeSH KeywordsAmino Acid SequenceBinding SitesCell LineChloridesCopperCross-Linking ReagentsDisulfidesHomeostasisHumansIon TransportIonsKineticsMicroscopy, ConfocalMicroscopy, FluorescenceModels, MolecularMolecular Sequence DataMutationPhenanthrolinesPhosphorylationProtein Structure, TertiaryRubidium RadioisotopesSequence Homology, Amino AcidSodium-Potassium-Chloride SymportersSolute Carrier Family 12, Member 2ConceptsTransmembrane domain 10Domain 10Copper phenanthrolineLarge C terminusNa-K-Cl cotransporterRegulatory domainCysteine pairsC-terminusN-terminusDephosphorylation rateTransporter activationDisulfide formationIon transportHomology modelNKCC activationInhibition of transportTransport functionLow micromolar concentrationsSame transporterCross-link formationActivation statePhosphorylationTerminusTransportersOcclusion step
2002
Functional comparison of renal Na-K-Cl cotransporters between distant species
Gagnon E, Forbush B, Caron L, Isenring P. Functional comparison of renal Na-K-Cl cotransporters between distant species. American Journal Of Physiology - Cell Physiology 2002, 284: c365-c370. PMID: 12388059, DOI: 10.1152/ajpcell.00262.2002.Peer-Reviewed Original ResearchConceptsRenal Na-K-Cl cotransporterNa-K-Cl cotransporterSplice variantsSecond transmembrane domainDistant vertebratesDistant speciesTransmembrane domainAlternative splicingXenopus laevis oocytesTransport activityCl(-) affinityRenal NKCC2Functional comparisonLaevis oocytesSpeciesSharksSimilar affinityIon dependenceA Regulatory Locus of Phosphorylation in the N Terminus of the Na-K-Cl Cotransporter, NKCC1*
Darman RB, Forbush B. A Regulatory Locus of Phosphorylation in the N Terminus of the Na-K-Cl Cotransporter, NKCC1*. Journal Of Biological Chemistry 2002, 277: 37542-37550. PMID: 12145304, DOI: 10.1074/jbc.m206293200.Peer-Reviewed Original ResearchMeSH KeywordsAmino Acid SequenceAnimalsBinding SitesCell LineDogfishHumansIon TransportKineticsModels, MolecularPeptide FragmentsPhosphopeptidesPhosphorylationProtein ConformationRecombinant ProteinsSalt GlandSodium-Potassium-Chloride SymportersSolute Carrier Family 12, Member 2Spectrometry, Mass, Matrix-Assisted Laser Desorption-IonizationTransfectionTrypsinConceptsProtein phosphatase 1 bindsN-terminusSer/ThrPhosphorylation-dependent mechanismN-terminal domainStrong consensus siteNa-K-Cl cotransporter NKCC1Matrix-assisted laser desorption ionization timePhosphoregulatory mechanismsPhosphoacceptor sitesRegulatory lociHEK-293 cellsNa-K-Cl cotransporterLaser desorption ionization timeCalyculin AConsensus sitesPhosphorylation stoichiometryDesorption ionization timeAmino acidsTryptic fragmentsProteinGland tubulesRectal gland tubulesFlight mass spectrometryIonization timeActivation of the Na-K-Cl Cotransporter NKCC1 Detected with a Phospho-specific Antibody*
Flemmer AW, Giménez I, Dowd BF, Darman RB, Forbush B. Activation of the Na-K-Cl Cotransporter NKCC1 Detected with a Phospho-specific Antibody*. Journal Of Biological Chemistry 2002, 277: 37551-37558. PMID: 12145305, DOI: 10.1074/jbc.m206294200.Peer-Reviewed Original ResearchMeSH KeywordsAmino Acid SequenceAnimalsAntibody SpecificityBinding SitesColonDipeptidesEpinephrineHumansIon TransportIsoproterenolKineticsMolecular Sequence DataMutagenesis, Site-DirectedPhosphatesPhosphopeptidesRatsRecombinant ProteinsSodium-Potassium-Chloride SymportersSolute Carrier Family 12, Member 2ThreonineTracheaConceptsPhospho-specific antibodiesNa-K-Cl cotransporter NKCC1Divergent vertebrate speciesBasolateral membraneShark rectal glandPhosphorylation of NKCC1Regulatory lociHEK-293 cellsHigh conservationThreonine residuesPhosphorylation experimentsVertebrate speciesRegulatory domainCotransporter NKCC1NKCC1 regulationN-terminusRectal glandCell membranePhosphorylationImmunofluorescence analysisAgonist stimulationActivation stateGland tubulesRectal gland tubulesEpithelial cells
1998
Mutagenic Mapping of the Na-K-Cl Cotransporter for Domains Involved in Ion Transport and Bumetanide Binding
Isenring P, Jacoby S, Chang J, Forbush B. Mutagenic Mapping of the Na-K-Cl Cotransporter for Domains Involved in Ion Transport and Bumetanide Binding. The Journal Of General Physiology 1998, 112: 549-558. PMID: 9806964, PMCID: PMC2229443, DOI: 10.1085/jgp.112.5.549.Peer-Reviewed Original ResearchMeSH KeywordsAnimalsBinding SitesBumetanideCarrier ProteinsCells, CulturedChloridesDiureticsHumansKidneyKineticsMutagenesis, Site-DirectedOligonucleotide ProbesPotassiumProtein Structure, TertiaryRecombinant Fusion ProteinsRubidium RadioisotopesSharksSodiumSodium-Potassium-Chloride SymportersSpecies SpecificityComparison of Na-K-Cl Cotransporters NKCC1, NKCC2, AND THE HEK CELL Na-K-Cl COTRANSPORTER*
Isenring P, Jacoby S, Payne J, Forbush B. Comparison of Na-K-Cl Cotransporters NKCC1, NKCC2, AND THE HEK CELL Na-K-Cl COTRANSPORTER*. Journal Of Biological Chemistry 1998, 273: 11295-11301. PMID: 9556622, DOI: 10.1074/jbc.273.18.11295.Peer-Reviewed Original ResearchConceptsNa-K-Cl cotransporterHEK-293 cellsNa-K-Cl cotransporter NKCC1Epithelial cellsBasolateral Cl- exitHEK-293Reverse transcriptase-polymerase chain reactionNa-K-ClRenal epithelial cellsEpithelial Cl- transportResting level of activityMaintenance of cell volumeSecretory epithelial cellsInhibitor bumetanideCotransporter NKCC1NKCC2 activityCl- exitNKCC1Cl- transportNKCC2CotransporterVolume responseNa affinityResting levelBumetanide
1997
Diversity of the E2P Phosphoforms of Na, K‐ATPasea
FEDOSOVA N, CORNELIUS F, FORBUSH B, KLODOS I. Diversity of the E2P Phosphoforms of Na, K‐ATPasea. Annals Of The New York Academy Of Sciences 1997, 834: 386-389. PMID: 9432913, DOI: 10.1111/j.1749-6632.1997.tb52278.x.Peer-Reviewed Original ResearchIon and Bumetanide Binding by the Na-K-Cl Cotransporter
Isenring P, Forbush B. Ion and Bumetanide Binding by the Na-K-Cl Cotransporter. Journal Of Biological Chemistry 1997, 272: 24556-24562. PMID: 9305921, DOI: 10.1074/jbc.272.39.24556.Peer-Reviewed Original ResearchConceptsNa-K-Cl cotransporterNa-K-ClBumetanide-sensitive 86Rb influxCation-chloride cotransporter familyApparent affinityHEK-293 cellsInhibitor bumetanideCell surface deliveryElectrolyte secretionTime course of activationBumetanidePolarized epitheliaBumetanide bindingCotransporterCl- mediumCotransported ionsHEK-293SNKCC1Hydrophobic central domainSurface deliveryCourse of activationCentral domainTime courseCytoplasmic N-Chimera approach
1996
Molecular Cloning and Functional Expression of the K-Cl Cotransporter from Rabbit, Rat, and Human A NEW MEMBER OF THE CATION-CHLORIDE COTRANSPORTER FAMILY*
Gillen C, Brill S, Payne J, Forbush B. Molecular Cloning and Functional Expression of the K-Cl Cotransporter from Rabbit, Rat, and Human A NEW MEMBER OF THE CATION-CHLORIDE COTRANSPORTER FAMILY*. Journal Of Biological Chemistry 1996, 271: 16237-16244. PMID: 8663127, DOI: 10.1074/jbc.271.27.16237.Peer-Reviewed Original ResearchMeSH KeywordsAmino Acid SequenceAnimalsBase SequenceBrainCarrier ProteinsCell LineChloridesCloning, MolecularDatabases, FactualDNA PrimersDNA, ComplementaryHumansKidneyKineticsModels, StructuralMolecular Sequence DataMultigene FamilyOpen Reading FramesPhylogenyPolymerase Chain ReactionPotassiumProtein Structure, SecondaryRabbitsRatsRecombinant ProteinsRubidiumSequence Homology, Amino AcidSequence Tagged SitesSymportersTransfectionConceptsNa-K-Cl cotransporterK-Cl cotransportK-ClRat brain cDNA libraryThiazide-sensitive Na-Cl cotransporterRate of 86Rb effluxBrain cDNA libraryBumetanide-sensitive Na-K-Cl cotransporterN-linked glycosylation sitesC-terminal regionSubstantial sequence homologyNa-Cl cotransporterNa-K-ClHEK 293) cellsUptake of 86RbNorthern blot analysisGenome sequenceCDNA libraryHydropathy analysisFunctional expressionSequence homologyReverse transcription-polymerase chain reactionHuman embryonic kidneySequence analysisHomologous cDNA probes
1994
Kinetic heterogeneity of phosphoenzyme of Na,K-ATPase modeled by unmixed lipid phases. Competence of the phosphointermediate.
Klodos I, Post R, Forbush B. Kinetic heterogeneity of phosphoenzyme of Na,K-ATPase modeled by unmixed lipid phases. Competence of the phosphointermediate. Journal Of Biological Chemistry 1994, 269: 1734-1743. PMID: 8294422, DOI: 10.1016/s0021-9258(17)42089-8.Peer-Reviewed Original Research
1992
The Na-K-Cl cotransporter of avian salt gland. Phosphorylation in response to cAMP-dependent and calcium-dependent secretogogues.
Torchia J, Lytle C, Pon D, Forbush B, Sen A. The Na-K-Cl cotransporter of avian salt gland. Phosphorylation in response to cAMP-dependent and calcium-dependent secretogogues. Journal Of Biological Chemistry 1992, 267: 25444-25450. PMID: 1281159, DOI: 10.1016/s0021-9258(19)74061-7.Peer-Reviewed Original ResearchMeSH Keywords1-Methyl-3-isobutylxanthine8-Bromo Cyclic Adenosine MonophosphateAnimalsBlotting, WesternCalciumCarbacholCarrier ProteinsCell MembraneColforsinCyclic AMPDucksKineticsMembrane ProteinsPhosphatesPhosphorylationSalt GlandSodium-Potassium-Chloride SymportersTime FactorsVasoactive Intestinal PeptideConceptsNa-K-Cl cotransporterPhosphorylation of pp170Avian salt glandMonoclonal antibodiesResponse to carbacholEffect of forskolinNa-K-ClIntact cell preparationsConcentration-dependent increaseStimulation of secretionIncreased approximately 5-foldCAMP accumulationExposure of cellsCAMP-dependent protein kinasePp170Approximately 5-foldCell preparationsCAMP-dependentSalt gland cellsGland membranesWestern blottingForskolinSecretogoguesVIPAntibodiesThe Na-K-Cl cotransport protein of shark rectal gland. II. Regulation by direct phosphorylation.
Lytle C, Forbush B. The Na-K-Cl cotransport protein of shark rectal gland. II. Regulation by direct phosphorylation. Journal Of Biological Chemistry 1992, 267: 25438-25443. PMID: 1334094, DOI: 10.1016/s0021-9258(19)74060-5.Peer-Reviewed Original ResearchMeSH KeywordsAmino Acid SequenceAnimalsCarrier ProteinsChromatography, High Pressure LiquidCyclic AMPDogfishHypertonic SolutionsIsoquinolinesKineticsMembrane ProteinsMolecular Sequence DataPeptide FragmentsPhosphorylationProtein Kinase InhibitorsRectumSebaceous GlandsSodium-Potassium-Chloride SymportersSulfanilamidesThionucleotidesConceptsNa-K-Cl cotransport proteinNa-K-ClCotransporter proteinShark rectal glandCell shrinkageCAMP-dependent secretagoguesRectal gland tubulesOsmotically induced changesCotransporter phosphorylationCotransport activityKinase inhibitorsProtein kinase inhibitorsCotransporterPhosphorylation stateRectal glandActive stateH-8K-252aTubulesGlandPhosphorylationCDNA analysisCellsIncreased 5Protein phosphorylation
1991
[3H]bumetanide binding to mouse kidney membranes: identification of corresponding membrane proteins
Haas M, Dunham P, Forbush B. [3H]bumetanide binding to mouse kidney membranes: identification of corresponding membrane proteins. American Journal Of Physiology 1991, 260: c791-c804. PMID: 2018111, DOI: 10.1152/ajpcell.1991.260.4.c791.Peer-Reviewed Original ResearchMeSH KeywordsAffinity LabelsAnimalsBenzophenonesBumetanideCarcinoma, Ehrlich TumorCarrier ProteinsCell FractionationCell MembraneCentrifugation, Density GradientFemaleKidneyKidney CortexKidney MedullaKineticsMembrane ProteinsMiceProtein BindingSodium-Potassium-Chloride SymportersSulfanilamidesTritiumConceptsMouse kidney membranesKidney membranesNa-K-Cl cotransport systemNa-K-ClLow-affinity peakDog kidney membraneMouse Ehrlich ascites tumor cellsMouse kidney proteinsDog kidneyCrude plasma membranesTumor cellsWestern blot analysisEhrlich ascites tumor cellsCotransport systemAscites tumor cellsMouse kidneyMiceStaining profileLow-affinity sitesWestern blottingBinding sitesAntiserum cross-reactedKidneyBlot analysisKidney proteins
1990
Biosynthesis of the Na,K-ATPase in Madin-Darby canine kidney cells. Activation and cell surface delivery.
Caplan MJ, Forbush B, Palade GE, Jamieson JD. Biosynthesis of the Na,K-ATPase in Madin-Darby canine kidney cells. Activation and cell surface delivery. Journal Of Biological Chemistry 1990, 265: 3528-3534. PMID: 2154482, DOI: 10.1016/s0021-9258(19)39801-1.Peer-Reviewed Original Research
1988
Rapid 86Rb release from an occluded state of the Na,K-pump reflects the rate of dephosphorylation or dearsenylation.
Forbush B. Rapid 86Rb release from an occluded state of the Na,K-pump reflects the rate of dephosphorylation or dearsenylation. Journal Of Biological Chemistry 1988, 263: 7961-7969. PMID: 2836403, DOI: 10.1016/s0021-9258(18)68428-5.Peer-Reviewed Original ResearchRapid release of 45Ca from an occluded state of the Na,K-pump.
Forbush B. Rapid release of 45Ca from an occluded state of the Na,K-pump. Journal Of Biological Chemistry 1988, 263: 7970-7978. PMID: 2836404, DOI: 10.1016/s0021-9258(18)68429-7.Peer-Reviewed Original ResearchConceptsNa,K-pumpNa,K-ATPaseK-pumpK-ATPasePhosphorylation of Na,K-ATPaseRelease of 45CaExposure to K+Release of 86RbApparent affinityCa2+Transport sitesK+ congenerMg2+ + ATPIntracellular faceRelease of K+Simultaneous occlusionExtracellular sitesPrevent phosphorylationCa2Exposure to Mg2Extracellular faceReleaseAbsence of PiN-methylglucamineIntracellular medium
1987
Rapid release of 42K and 86Rb from an occluded state of the Na,K-pump in the presence of ATP or ADP.
Forbush B. Rapid release of 42K and 86Rb from an occluded state of the Na,K-pump in the presence of ATP or ADP. Journal Of Biological Chemistry 1987, 262: 11104-11115. PMID: 2440883, DOI: 10.1016/s0021-9258(18)60932-9.Peer-Reviewed Original ResearchPhysiology and biophysics of chloride and cation cotransport across cell membranes.
Lauf P, McManus T, Haas M, Forbush B, Duhm J, Flatman P, Saier M, Russell J. Physiology and biophysics of chloride and cation cotransport across cell membranes. The FASEB Journal 1987, 46: 2377-94. PMID: 3552736.Peer-Reviewed Original Research
1984
Na+ movement in a single turnover of the Na pump.
Forbush B. Na+ movement in a single turnover of the Na pump. Proceedings Of The National Academy Of Sciences Of The United States Of America 1984, 81: 5310-5314. PMID: 6089192, PMCID: PMC391693, DOI: 10.1073/pnas.81.17.5310.Peer-Reviewed Original ResearchAn apparatus for rapid kinetic analysis of isotopic efflux from membrane vesicles and of ligand dissociation from membrane proteins
Forbush B. An apparatus for rapid kinetic analysis of isotopic efflux from membrane vesicles and of ligand dissociation from membrane proteins. Analytical Biochemistry 1984, 140: 495-505. PMID: 6091496, DOI: 10.1016/0003-2697(84)90200-8.Peer-Reviewed Original Research