2022
Structural basis for inhibition of the Cation-chloride cotransporter NKCC1 by the diuretic drug bumetanide
Zhao Y, Roy K, Vidossich P, Cancedda L, De Vivo M, Forbush B, Cao E. Structural basis for inhibition of the Cation-chloride cotransporter NKCC1 by the diuretic drug bumetanide. Nature Communications 2022, 13: 2747. PMID: 35585053, PMCID: PMC9117670, DOI: 10.1038/s41467-022-30407-3.Peer-Reviewed Original ResearchConceptsTranslocation pathwayElectron cryo-microscopy structureC-terminal domainIon translocation pathwayCation-chloride cotransporters NKCC1Transmembrane domainCotransporter NKCC1C-terminal domain interactionsStructural basisDomain interactionsRenal salt reabsorptionDomain associationConformational changesFunctional studiesIon translocationElectroneutral symportCell membraneNKCC1PathwayNKCC2DomainSalt reabsorptionTransmembraneTranslocationTransporters
2021
The structural basis of function and regulation of neuronal cotransporters NKCC1 and KCC2
Zhang S, Zhou J, Zhang Y, Liu T, Friedel P, Zhuo W, Somasekharan S, Roy K, Zhang L, Liu Y, Meng X, Deng H, Zeng W, Li G, Forbush B, Yang M. The structural basis of function and regulation of neuronal cotransporters NKCC1 and KCC2. Communications Biology 2021, 4: 226. PMID: 33597714, PMCID: PMC7889885, DOI: 10.1038/s42003-021-01750-w.Peer-Reviewed Original ResearchConceptsCryo-electron microscopy structureHuman NKCC1Microscopy structureEssential residuesFunctional characterizationKCC transportersPlasma membraneStructural basisTransepithelial saltTransport activityMechanistic understandingTransportersStructural studiesCritical roleCotransporter NKCC1Computational analysisIon transportWater transportNeuronal excitabilityNKCC1PhosphorylationCell volumeNKCCKCC2Residues
2010
Drosophila glia use a conserved cotransporter mechanism to regulate extracellular volume
Leiserson WM, Forbush B, Keshishian H. Drosophila glia use a conserved cotransporter mechanism to regulate extracellular volume. Glia 2010, 59: 320-332. PMID: 21125654, PMCID: PMC3005002, DOI: 10.1002/glia.21103.Peer-Reviewed Original ResearchMeSH KeywordsAction PotentialsAnimalsAnimals, Genetically ModifiedBlood-Nerve BarrierCells, CulturedDrosophilaDrosophila ProteinsExtracellular SpaceHumansIn Vitro TechniquesLarvaMicroscopy, Electron, TransmissionModels, BiologicalMutationNeural ConductionNeurogliaNeuronsPeripheral NervesProtein Serine-Threonine KinasesSymportersTwo-Hybrid System TechniquesConceptsHuman NKCC1Yeast two hybrid assaysExtracellular solute compositionLarvae mutantDrosophila gliaNcc69Osmotic homeostasisExtracellular volumeMolecular mechanismsNervous systemOrthologsExtracellular solutesPhysiological mechanismsBlood-brain barrierBlood-nerve barrierSimilar roleAccumulation of fluidAction potential conductionGlial cellsPeripheral neuropathyNKCC1Serious health threatDetectable impactBlood barrierGlia
2008
Na-K-Cl Cotransporter-1 in the Mechanism of Ammonia-induced Astrocyte Swelling*
Jayakumar A, Liu M, Moriyama M, Ramakrishnan R, Forbush B, Reddy P, Norenberg M. Na-K-Cl Cotransporter-1 in the Mechanism of Ammonia-induced Astrocyte Swelling*. Journal Of Biological Chemistry 2008, 283: 33874-33882. PMID: 18849345, PMCID: PMC2590687, DOI: 10.1074/jbc.m804016200.Peer-Reviewed Original ResearchConceptsNKCC activityNa-K-ClAstrocyte swellingCell swellingNa-K-Cl cotransporter-1Na-K-Cl cotransporterComplication of acute liver failureN-nitro-L-arginine methyl esterNitric oxide synthase inhibitionActivation of NKCC1NKCC1 protein expressionCultured astrocytesInduce oxidative/nitrosative stressAcute liver failureLoss of ion homeostasisSwelling of astrocytesPhosphorylated NKCC1NKCC1 expressionLiver failureNKCC1Synthase inhibitionBrain herniationBrain edemaIntracranial pressureProtein expression
2006
Expression of the basolateral Na–K–Cl cotransporter during mouse nephrogenesis and embryonic development
Heuvel G, Payne J, Igarashi P, Forbush B. Expression of the basolateral Na–K–Cl cotransporter during mouse nephrogenesis and embryonic development. Gene Expression Patterns 2006, 6: 1000-1006. PMID: 16814616, DOI: 10.1016/j.modgep.2006.04.004.Peer-Reviewed Original ResearchConceptsIn situ hybridizationNa-K-Cl cotransporterDorsal root ganglionBranching ureteric budEndocapillary cellsPeripheral nervous systemRoot ganglionTooth budsNasal epitheliumMouse nephrogenesisChoroid plexusSubmandibular glandMature nephronsUreteric budNorthern blot analysisSLC12A2Nervous systemMesenchymal cellsNKCC1Blot analysisMetanephroiNephric structuresGlomeruliMiceEmbryonic development
2003
PASK (Proline-Alanine-rich STE20-related Kinase), a Regulatory Kinase of the Na-K-Cl Cotransporter (NKCC1)*
Dowd BF, Forbush B. PASK (Proline-Alanine-rich STE20-related Kinase), a Regulatory Kinase of the Na-K-Cl Cotransporter (NKCC1)*. Journal Of Biological Chemistry 2003, 278: 27347-27353. PMID: 12740379, DOI: 10.1074/jbc.m301899200.Peer-Reviewed Original ResearchMeSH KeywordsAnimalsBlotting, WesternCell LineDNA, ComplementaryDose-Response Relationship, DrugGenes, DominantGenetic VectorsHumansMarine ToxinsOxazolesOxidative StressPhosphorylationPrecipitin TestsProtein BindingProtein Serine-Threonine KinasesProtein Structure, TertiaryRatsRubidiumSharksSodium-Potassium-Chloride SymportersSolute Carrier Family 12, Member 2ThreonineTime FactorsConceptsNa-K-Cl cotransporterN-terminal regulatory domainPhosphorylation-dependent activationHEK cellsInhibitor calyculin ANKCC1 activityRegulatory domainCoimmunoprecipitation assaysRegulatory kinasesActivation of NKCC1Calyculin ARegulated eventNKCC1 activationPhosphorylationKinaseSharksCotransporter activityOverexpressionCotransporter expressionNKCC1CellsActivationBindingCotransporterMutants
2000
The Na-K-Cl Cotransporter of Secretory Epithelia
Haas M, Forbush B. The Na-K-Cl Cotransporter of Secretory Epithelia. Annual Review Of Physiology 2000, 62: 515-534. PMID: 10845101, DOI: 10.1146/annurev.physiol.62.1.515.Peer-Reviewed Original ResearchConceptsNa-K-ClNa-K-Cl cotransporterRegulation of NKCC1Secretory epitheliaNa-K-Cl cotransporter isoformsK-Cl cotransporter isoformsApical Cl- channelsNa-Cl cotransporterNon-epithelial cellsNKCC1 regulationCl- channelsBasolateral membraneCotransporter proteinFluid secretionNKCC isoformsNKCC1Ascending limbHenle's loopEpithelial cellsMacula densaIon transport proteinsTransport NaEpitheliaCotransporterCell volume
1998
Comparison 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