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
2012
Loop Diuretic and Ion-binding Residues Revealed by Scanning Mutagenesis of Transmembrane Helix 3 (TM3) of Na-K-Cl Cotransporter (NKCC1)*
Somasekharan S, Tanis J, Forbush B. Loop Diuretic and Ion-binding Residues Revealed by Scanning Mutagenesis of Transmembrane Helix 3 (TM3) of Na-K-Cl Cotransporter (NKCC1)*. Journal Of Biological Chemistry 2012, 287: 17308-17317. PMID: 22437837, PMCID: PMC3366785, DOI: 10.1074/jbc.m112.356014.Peer-Reviewed Original ResearchConceptsTransmembrane helix 3Na-K-Cl cotransporterTranslocation pathwayHelix 3Homology modelTryptophan-scanning mutagenesisMutation of residuesStructural homology modelEpithelial salt transportExtracellular gateCellular chloride homeostasisScanning mutagenesisOpen conformationIntracellular endPore residuesFunctional roleIon translocationTranslocation rateResiduesMutagenesisCentral roleChloride homeostasisMutationsPathwayLarge effect
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 Specificity
1997
Ion 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
1995
Primary 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 cDNA
1994
Molecular 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
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
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
1987
Na,K,Cl-cotransport system: characterization by bumetanide binding and photolabelling.
Haas M, Forbush B. Na,K,Cl-cotransport system: characterization by bumetanide binding and photolabelling. Kidney International Supplement 1987, 23: s134-43. PMID: 3481640.Peer-Reviewed Original ResearchPhotolabeling of a 150-kDa (Na + K + Cl) cotransport protein from dog kidney with a bumetanide analogue
Haas M, Forbush B. Photolabeling of a 150-kDa (Na + K + Cl) cotransport protein from dog kidney with a bumetanide analogue. American Journal Of Physiology 1987, 253: c243-c252. PMID: 3618761, DOI: 10.1152/ajpcell.1987.253.2.c243.Peer-Reviewed Original ResearchConceptsKidney cortexBinding to kidney membranesDog kidney cortexSite of actionCotransporter proteinApical membraneAscending limbBumetanideCotransport systemHenle's loopEpithelial cellsPhotolysis mediumKidney membranesK 1/2Mammalian kidneyCrude membranesBenzophenone analoguesKidneyDog kidneyPresence of NaCortexPhotolysisSucrose density gradientsMolecular mass proteinsDogsPhysiology 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
1986
[3H]bumetanide binding to duck red cells. Correlation with inhibition of (Na + K + 2Cl) co-transport.
Haas M, Forbush B. [3H]bumetanide binding to duck red cells. Correlation with inhibition of (Na + K + 2Cl) co-transport. Journal Of Biological Chemistry 1986, 261: 8434-8441. PMID: 3013852, DOI: 10.1016/s0021-9258(19)83931-5.Peer-Reviewed Original ResearchConceptsDuck red cellsExtracellular sodiumRed cellsCo-transportDog kidney outer medullaBinding of [3H]bumetanideConcentration of extracellular sodiumIncreased extracellular sodiumPresence of norepinephrineCo-transport systemBumetanide inhibitionKidney outer medullaOuter medullaBiphasic effectInhibition of influxCell shrinkageHalf-maximalSaturable bindingBumetanideInhibit bindingCell typesSaturable mannerNorepinephrineInfluxBinding to membranes
1983
[3H]bumetanide binding to membranes isolated from dog kidney outer medulla. Relationship to the Na,K,Cl co-transport system.
Forbush B, Palfrey H. [3H]bumetanide binding to membranes isolated from dog kidney outer medulla. Relationship to the Na,K,Cl co-transport system. Journal Of Biological Chemistry 1983, 258: 11787-11792. PMID: 6619143, DOI: 10.1016/s0021-9258(17)44299-2.Peer-Reviewed Original ResearchConceptsOuter medullaCl- co-transport systemDog kidney outer medullaCl- cotransport systemLoop of HenleCo-transport systemLoop diureticsBasolateral membraneKidney outer medullaProximal tubulesAscending limbLuminal membraneCotransport systemKidney cortexMedullaKidney membranesBrush borderCo-transportCrude membranesKidneyDog kidneyBinding to membranesBinding activityGradient centrifugation