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
2002
Activation 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
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 fragments
1988
Photoaffinity labelling of a 150 kDa (Na + K + Cl)-cotransport protein from duck red cells with an analog of bumetanide
Haas M, Forbush B. Photoaffinity labelling of a 150 kDa (Na + K + Cl)-cotransport protein from duck red cells with an analog of bumetanide. Biochimica Et Biophysica Acta 1988, 939: 131-144. PMID: 3349075, DOI: 10.1016/0005-2736(88)90054-5.Peer-Reviewed Original ResearchConceptsDuck red cellsRed cellsSimultaneous presence of Na+Binding to intact cellsIncubation mediumDog kidney membraneConsistent with labelingDuck red blood cellsRed blood cellsBumetanideCotransporterRed cell membraneBlood cellsKidney membranesPresence of Na+DucksCellsCell membranePlasma membraneSDS-polyacrylamide gelsIntact cellsSDS-polyacrylamideProteinNorepinephrine