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
2009
Sites of Regulated Phosphorylation that Control K-Cl Cotransporter Activity
Rinehart J, Maksimova YD, Tanis JE, Stone KL, Hodson CA, Zhang J, Risinger M, Pan W, Wu D, Colangelo CM, Forbush B, Joiner CH, Gulcicek EE, Gallagher PG, Lifton RP. Sites of Regulated Phosphorylation that Control K-Cl Cotransporter Activity. Cell 2009, 138: 525-536. PMID: 19665974, PMCID: PMC2811214, DOI: 10.1016/j.cell.2009.05.031.Peer-Reviewed Original ResearchMeSH KeywordsAmino Acid SequenceAmino Acid SubstitutionAnimalsHumansMiceMolecular Sequence DataPhosphorylationSequence AlignmentSymportersConceptsIntrinsic transport activityK-Cl cotransporterTransport activityCell volume regulationRegulated phosphorylationRNA interferenceAlanine substitutionsCultured cellsHomologous sitesKCC activityCl exitWNK1 expressionNeonatal mouse brainVolume regulationNeuronal functionHypotonic conditionsActive cotransportPhosphorylationIntracellular chloride concentrationCotransporter activityKCC3Human red blood cellsKCC2 activationFundamental roleMouse brain
2005
Regulatory phosphorylation sites in the NH2 terminus of the renal Na-K-Cl cotransporter (NKCC2)
Giménez I, Forbush B. Regulatory phosphorylation sites in the NH2 terminus of the renal Na-K-Cl cotransporter (NKCC2). American Journal Of Physiology. Renal Physiology 2005, 289: f1341-f1345. PMID: 16077079, DOI: 10.1152/ajprenal.00214.2005.Peer-Reviewed Original Research
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 cellsSpatially Distributed Alternative Splice Variants of the Renal Na-K-Cl Cotransporter Exhibit Dramatically Different Affinities for the Transported Ions*
Giménez I, Isenring P, Forbush B. Spatially Distributed Alternative Splice Variants of the Renal Na-K-Cl Cotransporter Exhibit Dramatically Different Affinities for the Transported Ions*. Journal Of Biological Chemistry 2002, 277: 8767-8770. PMID: 11815599, DOI: 10.1074/jbc.c200021200.Peer-Reviewed Original Research
1998
The role of transmembrane domain 2 in cation transport by the Na–K–Cl cotransporter
Isenring P, Jacoby S, Forbush B. The role of transmembrane domain 2 in cation transport by the Na–K–Cl cotransporter. Proceedings Of The National Academy Of Sciences Of The United States Of America 1998, 95: 7179-7184. PMID: 9618559, PMCID: PMC22778, DOI: 10.1073/pnas.95.12.7179.Peer-Reviewed Original ResearchConceptsNa-K-Cl cotransporterBumetanide-sensitive 86Rb influxTransmembrane domain 2Na-Cl cotransporterBumetanide bindingNa-K-ClHEK-293 cellsHuman cotransportersDomain 2Third transmembrane domainCotransporterNa affinityHEK-293Stable transfectionNKCC2BumetanideCation transportTransmembrane domain 3Transmembrane domainDomain 3Alternative splicingIon transport
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
1995
Cloning, embryonic expression, and alternative splicing of a murine kidney-specific Na-K-Cl cotransporter
Igarashi P, Heuvel G, Payne J, Forbush B. Cloning, embryonic expression, and alternative splicing of a murine kidney-specific Na-K-Cl cotransporter. American Journal Of Physiology 1995, 269: f405-f418. PMID: 7573490, DOI: 10.1152/ajprenal.1995.269.3.f405.Peer-Reviewed Original ResearchConceptsNa-K-Cl cotransporterExpression of NKCC2In situ hybridizationCassette exonsRenal Na-K-Cl cotransporterOuter medullaAscending limbAlternative splicingInner stripe of outer medullaPotential sites of phosphorylationAdenosine 3',5'-cyclic monophosphate-dependent protein kinaseDays post coitusFull-length cDNAAnchored polymerase chain reactionSites of phosphorylationConsensus splice donorLoop of HenleS-shaped bodiesProtein kinase CNorthern blot analysisPattern of expressionGenomic clonesNKCC2Outer stripeProtein sequencesPrimary Structure, Functional Expression, and Chromosomal Localization of the Bumetanide-sensitive Na-K-Cl Cotransporter in Human Colon *
Payne J, Xu J, Haas M, Lytle C, Ward D, Forbush B. Primary Structure, Functional Expression, and Chromosomal Localization of the Bumetanide-sensitive Na-K-Cl Cotransporter in Human Colon *. Journal Of Biological Chemistry 1995, 270: 17977-17985. PMID: 7629105, DOI: 10.1074/jbc.270.30.17977.Peer-Reviewed Original ResearchMeSH KeywordsAmino Acid SequenceAnimalsBlotting, NorthernBumetanideCarrier ProteinsCell LineChloridesChromosome MappingChromosomes, Human, Pair 5Cloning, MolecularColonDNA, ComplementaryHumansMolecular Sequence DataPotassiumProtein ConformationSodiumSodium-Potassium-Chloride SymportersTumor Cells, CulturedConceptsNa-K-Cl cotransporterT84 cellsRenal Na-K-Cl cotransporterThiazide-sensitive Na-Cl cotransporterBumetanide-sensitive 86Rb influxBumetanide-sensitive Na-K-Cl cotransporterNa-Cl cotransporterHuman colon carcinoma lineNa-K-ClColon carcinoma linePrimary structureHuman embryonic kidney cellsChloride-free mediumShark rectal glandProtein kinase AStably transfected cellsG + C contentEmbryonic kidney cellsElectroneutral cotransporterUrinary bladderCarcinoma linesAbsorptive epitheliaSNKCC1Screening cDNA librariesBiotin-labeled cDNAMolecular characterization of the epithelial NaKCl cotransporter isoforms
Payne J, Forbush B. Molecular characterization of the epithelial NaKCl cotransporter isoforms. Current Opinion In Cell Biology 1995, 7: 493-503. PMID: 7495568, DOI: 10.1016/0955-0674(95)80005-0.Peer-Reviewed Original Research
1994
Alternatively spliced isoforms of the putative renal Na-K-Cl cotransporter are differentially distributed within the rabbit kidney.
Payne J, Forbush B. Alternatively spliced isoforms of the putative renal Na-K-Cl cotransporter are differentially distributed within the rabbit kidney. Proceedings Of The National Academy Of Sciences Of The United States Of America 1994, 91: 4544-4548. PMID: 7514306, PMCID: PMC43822, DOI: 10.1073/pnas.91.10.4544.Peer-Reviewed Original ResearchMeSH KeywordsAlternative SplicingAmino Acid SequenceAnimalsBase SequenceBlotting, NorthernCarrier ProteinsCloning, MolecularColonConserved SequenceDogfishGene LibraryHumansKidneyKidney CortexKidney MedullaMembrane ProteinsMolecular Sequence DataOpen Reading FramesPoly ARabbitsRNARNA, MessengerSalt GlandSequence Homology, Amino AcidSodium-Potassium-Chloride SymportersTranscription, GeneticConceptsNa-K-Cl cotransporterNorthern blot analysisRabbit kidney cDNA libraryRenal Na-K-Cl cotransporterC-terminal cytoplasmic domainC-terminal cytoplasmic regionConsensus splice acceptor siteKidney cDNA libraryAmino acid hydropathySplice acceptor siteBlot analysisN-linked glycosylationRegions of divergenceIntronic DNACDNA clonesCassette exonsCDNA libraryProtein kinase ACytoplasmic domainSpliced isoformsTransmembrane helicesCDNA probeCytoplasmic regionHomologous variantMultiple clonesMolecular cloning and functional expression of the bumetanide-sensitive Na-K-Cl cotransporter.
Xu J, Lytle C, Zhu T, Payne J, Benz E, Forbush B. Molecular cloning and functional expression of the bumetanide-sensitive Na-K-Cl cotransporter. Proceedings Of The National Academy Of Sciences Of The United States Of America 1994, 91: 2201-2205. PMID: 8134373, PMCID: PMC43338, DOI: 10.1073/pnas.91.6.2201.Peer-Reviewed Original ResearchConceptsNa-K-Cl cotransporterBumetanide-sensitive Na-K-Cl cotransporterNa-K-ClNa-K-Cl cotransport activityNa-K-Cl cotransport proteinThiazide-sensitive Na-Cl cotransporterCoupled movement of NaUncharacterized genesShort stretches of homologyRegulation of ionic balanceStretches of homologyC. elegans cDNAHEK-293 cellsHuman HEK-293 cellsPutative transmembrane domainsBumetanide sensitivityCotransport activityCotransporter proteinMovement of NaUrinary bladderAbsorptive epitheliaNorthern blot analysisPhosphoacceptor residuesCDNA sequenceCoding region
1993
Molecular cloning and immunological characterization of the gamma polypeptide, a small protein associated with the Na,K-ATPase.
Mercer R, Biemesderfer D, Bliss D, Collins J, Forbush B. Molecular cloning and immunological characterization of the gamma polypeptide, a small protein associated with the Na,K-ATPase. Journal Of Cell Biology 1993, 121: 579-586. PMID: 8387529, PMCID: PMC2119561, DOI: 10.1083/jcb.121.3.579.Peer-Reviewed Original ResearchConceptsNa,K-ATPaseGamma subunitK-ATPaseSmall membrane proteinsN-linked glycosylationTissue-specific fashionCardiac glycoside bindingGamma subunit mRNAGamma-specific antibodiesNorthern blot analysisHydropathy analysisSequenced proteinsNAB-ouabainMembrane proteinsGamma polypeptidesMolecular cloningGlycoside bindingSmall proteinsNephron segmentsBeta subunitSubunit mRNAAlpha subunitSubunitAmino acidsHydrophobic domains
1992
The Na-K-Cl cotransport protein of shark rectal gland. I. Development of monoclonal antibodies, immunoaffinity purification, and partial biochemical characterization.
Lytle C, Xu J, Biemesderfer D, Haas M, Forbush B. The Na-K-Cl cotransport protein of shark rectal gland. I. Development of monoclonal antibodies, immunoaffinity purification, and partial biochemical characterization. Journal Of Biological Chemistry 1992, 267: 25428-25437. PMID: 1460038, DOI: 10.1016/s0021-9258(19)74059-9.Peer-Reviewed Original ResearchMeSH KeywordsAmino Acid SequenceAnimalsAntibodies, MonoclonalBenzophenonesBlotting, WesternCarrier ProteinsCell MembraneChloridesCholic AcidsChromatography, AffinityChromatography, High Pressure LiquidChromatography, Ion ExchangeDogfishElectrophoresis, Polyacrylamide GelFluorescent Antibody TechniqueMembrane ProteinsMiceMice, Inbred BALB CMolecular Sequence DataMolecular WeightPeptide FragmentsPotassiumRectumSebaceous GlandsSodiumSodium-Potassium-Chloride SymportersSulfanilamidesConceptsNa-K-Cl cotransport proteinNa-K-ClCotransporter proteinNa-K-Cl cotransporterMonoclonal antibodiesShark rectal glandLoop diureticsBasolateral membraneApical membraneTransport of NaRepresentative antibodiesCotransporterCoupled transport of NaAntibodiesPattern of recognitionTreated with N-glycanaseGland secretory cellsImmunoelectron microscopyCell membranePlasma membraneEpitopesPhysiological evidenceSecretory cellsProteolytic fragmentsThe 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