2013
Functional Expression of Human NKCC1 from a Synthetic Cassette-Based cDNA: Introduction of Extracellular Epitope Tags and Removal of Cysteines
Somasekharan S, Monette MY, Forbush B. Functional Expression of Human NKCC1 from a Synthetic Cassette-Based cDNA: Introduction of Extracellular Epitope Tags and Removal of Cysteines. PLOS ONE 2013, 8: e82060. PMID: 24339991, PMCID: PMC3855340, DOI: 10.1371/journal.pone.0082060.Peer-Reviewed Original ResearchConceptsNa-K-Cl cotransporterEpitope tagYFP tagPlasma membraneMost animal cellsRemoval of cysteineOpen reading frameSilent restriction sitesHuman NKCC1Regulation of NKCC1Na-K-Cl cotransport activityHEK-293 cellsCellular homeostasisBiosynthetic machinerySynthetic cassetteAnimal cellsNovel cDNACysteine mutantsReading frameSynthetic cDNACysteine mutationsEndoplasmic reticulumFunctional expressionCotransport activityCDNA
1999
Functional interaction of the K-Cl cotransporter (KCC1) with the Na-K-Cl cotransporter in HEK-293 cells
Gillen C, Forbush B. Functional interaction of the K-Cl cotransporter (KCC1) with the Na-K-Cl cotransporter in HEK-293 cells. American Journal Of Physiology 1999, 276: c328-c336. PMID: 9950760, DOI: 10.1152/ajpcell.1999.276.2.c328.Peer-Reviewed Original ResearchConceptsNa-K-Cl cotransporterNa-K-Cl cotransport activityNa-K-ClK-Cl cotransportHEK-293 cellsCotransport activityK-ClHEK-293Activation of Na-K-Cl cotransportCell swellingK-Cl cotransport activityHypotonic cell swellingIntracellular Cl- concentrationFunctional interactionsKCC1External KCotransporterIon-transporting epitheliaCell linesN-ethylmaleimidePhysiological rangeCellsInvestigate functional interactionsCross-talkPretreatment
1998
The Na-K-Cl Cotransporters
Haas M, Forbush B. The Na-K-Cl Cotransporters. Journal Of Bioenergetics And Biomembranes 1998, 30: 161-172. PMID: 9672238, DOI: 10.1023/a:1020521308985.Peer-Reviewed Original ResearchConceptsNa-K-Cl cotransporterNa-K-ClNa-K-Cl cotransport activityNa-K-Cl cotransporter isoformsCotransport activitySecretory epitheliaApical Cl- channelsHypokalemic metabolic alkalosisCation-chloride cotransportersNKCC2 geneCl- channelsBartter's syndromeVolume depletionBasolateral membraneCotransporter proteinApical membraneSalt wastingMetabolic alkalosisSecreting epitheliaNonepithelial cellsAscending limbHenle's loopK-ClCotransporterEpithelial cells
1996
Na:K:2Cl Cotransporter (NKCC) of Intestinal Epithelial Cells SURFACE EXPRESSION IN RESPONSE TO cAMP*
D'Andrea L, Lytle C, Matthews J, Hofman P, Forbush B, Madara J. Na:K:2Cl Cotransporter (NKCC) of Intestinal Epithelial Cells SURFACE EXPRESSION IN RESPONSE TO cAMP*. Journal Of Biological Chemistry 1996, 271: 28969-28976. PMID: 8910547, DOI: 10.1074/jbc.271.46.28969.Peer-Reviewed Original ResearchConceptsChloride secretionSurface expressionBumetanide-sensitive NaMonoclonal antibodiesIntestinal chloride secretionIncreased surface expressionCell surface expressionEpithelial cell lineNKCC activityT84 monolayersCotransport activityAgonist stimulationCAMP agonistsEpithelial uptakeCotransporterBasolateral domainIntestinal epitheliaSurface biotinylationNKCCConfocal localizationApproximately 6-foldCell linesF-actin cytoskeletonSecretionImmunoelectron microscopyRegulatory phosphorylation of the secretory Na-K-Cl cotransporter: modulation by cytoplasmic Cl
Lytle C, Forbush B. Regulatory phosphorylation of the secretory Na-K-Cl cotransporter: modulation by cytoplasmic Cl. American Journal Of Physiology 1996, 270: c437-c448. PMID: 8779905, DOI: 10.1152/ajpcell.1996.270.2.c437.Peer-Reviewed Original ResearchConceptsNa-K-Cl cotransporterNa-K-ClCotransport activityNa-K-Cl cotransport proteinResponse to cell shrinkageIncreased cotransport activityExtracellular K concentrationBlocking activityRefractory to forskolinPhosphatase inhibitor calyculin AInhibitor calyculin AShark rectal glandCotransporter proteinCytoplasmic Cl-CotransporterCell ClSecretory stimuliCell shrinkageCalyculin APhosphorylationRate of entryRegulatory phosphorylationPhosphorylation stateSecretory tubulesRectal gland
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
1992
The 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