Affinage

Showing SLC12A1 — NKCC2 is a alias.

SLC12A1

Solute carrier family 12 member 1 · UniProt Q13621

Length
1099 aa
Mass
121.5 kDa
Annotated
2026-06-10
100 papers in source corpus 35 papers cited in narrative 35 extracted findings
Cross-family judge vs UniProt: Affinage preferred faithfulness: 7/7 claims corpus-supported (100%)

Mechanistic narrative

Synthesis pass · prose summary of the discoveries below

SLC12A1/NKCC2 is the apical Na-K-2Cl cotransporter of the renal thick ascending limb (TAL) that drives transepithelial NaCl reabsorption, and its loss-of-function mutations cause Bartter syndrome type I (PMID:8640224, PMID:12761241). The protein localizes exclusively to the apical membrane of medullary and cortical TAL segments (PMID:8853424), where it operates as a strictly ion-coupled carrier that—unlike its paralog NKCC1—does not cotransport water and is activated by low intracellular chloride (PMID:9556622, PMID:22250214). NKCC2 activity is governed by phosphorylation of conserved N-terminal threonines that constitute a regulatory phospho-domain required for the full hypertonic/low-chloride response (PMID:16077079, PMID:21321328); this is executed by a chloride-sensing kinase cascade in which WNK3 acts upstream of SPAK/OSR1, which dock on an RFQV motif and directly phosphorylate the cluster (PMID:16275913, PMID:18550832, PMID:21321328), while AMPK independently phosphorylates Ser126 to set basal isotonic activity (PMID:17341212). In vivo, kidney-specific WNK1 negatively regulates surface NKCC2 (PMID:21131289), and the dominant physiological stimulus is vasopressin acting through PKA—and adenylyl cyclase 6—to drive VAMP2-dependent exocytic insertion into the apical membrane (PMID:12732642, PMID:19592485, PMID:25008321, PMID:23123217). Surface abundance is further tuned by constitutive dynamin-2/clathrin/lipid-raft endocytosis and recycling (PMID:22977238, PMID:20719977) and by a series of interacting proteins controlling biogenesis and trafficking: conserved C-terminal di-leucine-like motifs (including LLV 1081-1083) mediate ER exit (PMID:19535327, PMID:23105100), OS9 routes immature ER-localized NKCC2 to glycan-dependent ERAD (PMID:26721884), SCAMP2 and aldolase B suppress surface delivery (PMID:17848580, PMID:21205824), MAL/VIP17 promotes membrane retention (PMID:20861303), and Nedd4-2 mediates ubiquitin-proteasome degradation (PMID:23104236). Additional modulators include Tamm-Horsfall protein, which facilitates chloride-sensitive activation (PMID:21737451), and nitric oxide, TNF-alpha, and IL-1R1 signaling that adjust NKCC2 expression and activity in blood-pressure regulation (PMID:26712462, PMID:12837683, PMID:26887831). Beyond the kidney, NKCC2 is expressed in the hypothalamo-neurohypophyseal system where it contributes to osmotically driven AVP neuron excitation and fluid balance (PMID:25834041).

Mechanistic history

Synthesis pass · year-by-year structured walk · 14 steps
  1. 1996 High

    Establishing that NKCC2 is genetically required for renal salt reabsorption answered whether this transporter is physiologically essential, linking the gene directly to human disease.

    Evidence Genetic linkage and mutation co-segregation in Bartter syndrome type I families, plus apical TAL immunolocalization in rat kidney

    PMID:8640224 PMID:8853424

    Open questions at the time
    • Did not define the transport mechanism or regulatory inputs
    • Functional consequences of individual mutations not yet tested
  2. 1998 High

    Heterologous characterization distinguished NKCC2 from NKCC1 kinetically, establishing its low-chloride activation and apical-matched transport behavior.

    Evidence Stable expression of rabbit NKCC2A chimera in HEK-293 cells with ion affinity and volume-response assays

    PMID:9556622

    Open questions at the time
    • Chimeric construct rather than full native human protein
    • Molecular basis of chloride sensing not resolved
  3. 2003 High

    Functional testing of Bartter mutations showed loss of transport despite membrane delivery, distinguishing trafficking defects from intrinsic functional impairment.

    Evidence Bumetanide-sensitive 22Na+ uptake in oocytes with immunoblotting and immunocytochemistry of six disease mutants

    PMID:12761241

    Open questions at the time
    • Did not establish structural basis of functional impairment per mutation
  4. 2005 High

    Identifying WNK3 as a positive regulator and the N-terminal threonine phospho-domain defined the activating signal input controlling NKCC2.

    Evidence Co-expression and mutagenesis in Xenopus oocytes with kinase-inactive WNK3 and threonine mutants under hypertonic stress

    PMID:16077079 PMID:16275913

    Open questions at the time
    • In vivo relevance of WNK3 not yet tested
    • Upstream chloride sensor not yet defined
  5. 2008 High

    Demonstrating chloride-depletion-driven phosphorylation requiring both WNK3 and SPAK established NKCC2 as the output of a chloride-sensing kinase cascade.

    Evidence Oocyte co-expression with chloride manipulation, phospho-site mutagenesis, kinase-inactive WNK3, and SPAK-binding motif deletion

    PMID:18550832

    Open questions at the time
    • Direct kinase-substrate phosphorylation not yet mapped biochemically
  6. 2009 High

    Defining the cAMP/PKA-driven exocytic mechanism and C-terminal ER-exit motifs explained how NKCC2 surface abundance is acutely and constitutively controlled.

    Evidence Surface biotinylation with selective PKA/Epac agonists and FM1-43 exocytosis in native TALs; pulse-chase and LLV-motif mutagenesis for ER exit

    PMID:19535327 PMID:19592485

    Open questions at the time
    • Identity of the SNARE machinery mediating exocytosis not yet resolved
    • Vesicle pool source unclear
  7. 2011 High

    Direct kinase-substrate mapping placed SPAK/OSR1 (via the RFQV motif) and a separate Ser130 kinase as the executors of activation, and distinguished NKCC2 from NCC in not requiring phosphorylation-triggered translocation.

    Evidence Mass spectrometry, phosphospecific antibodies, RFQV motif mutation, and isoform expression

    PMID:21321328

    Open questions at the time
    • Ser130 kinase identity not definitively established
    • Relative contributions of each site in vivo unclear
  8. 2011 High

    In vivo mouse models clarified the physiological regulators, showing KS-WNK1 negatively controls surface NKCC2 and THP facilitates chloride-sensitive activation.

    Evidence Transgenic and knockout mice with phospho-NKCC2 immunostaining; THP-KO mice, oocyte co-injection, and cultured TAL cells

    PMID:21131289 PMID:21737451

    Open questions at the time
    • Mechanism of THP-NKCC2 functional coupling not fully defined
  9. 2007 High

    Identifying AMPK-mediated Ser126 phosphorylation and aldolase B binding revealed metabolism-linked regulation of basal NKCC2 activity and surface levels.

    Evidence In vitro kinase assay, co-precipitation, and S126A oocyte expression; yeast two-hybrid, co-IP, and substrate-dependent disruption of aldolase B binding

    PMID:17341212 PMID:17848580

    Open questions at the time
    • In vivo importance of AMPK-Ser126 axis under physiological metabolic states not established
  10. 2010 High

    Dissecting endocytosis, recycling, and the MAL/VIP17 retention interaction established how internalization balances exocytosis to set steady-state surface NKCC2.

    Evidence Combinatorial endocytosis inhibitors and surface biotinylation in native THALs; co-IP and transgenic overexpression for MAL/VIP17

    PMID:20719977 PMID:20861303 PMID:22977238

    Open questions at the time
    • Adaptor proteins linking NKCC2 to clathrin not identified
    • MAL/VIP17 finding is Medium confidence from a single lab
  11. 2011 High

    Characterizing rare hypomorphic human variants and SCAMP2 binding linked NKCC2 trafficking and function to blood-pressure variation in the population.

    Evidence Heterologous expression of nine blood-pressure-associated variants; yeast two-hybrid and co-IP for SCAMP2 with E-peptide mutagenesis

    PMID:21205824 PMID:21209010

    Open questions at the time
    • Population-level penetrance of variant effects not addressed mechanistically
  12. 2012 High

    Resolving that NKCC2 does not cotransport water and defining TM2/ICL1 residues governing ion affinity clarified its transport mechanism distinct from NKCC1.

    Evidence Oocyte volume and 86Rb+ flux comparison of NKCC1 vs NKCC2; site-directed mutagenesis of TM2/ICL1 residues across splice variants

    PMID:17186942 PMID:22250214

    Open questions at the time
    • No high-resolution structure to confirm chloride-binding geometry
  13. 2015 High

    Identifying VAMP2 as the selective SNARE for cAMP/PKA exocytosis and OS9 as the ERAD route for immature NKCC2 completed the trafficking life-cycle picture.

    Evidence Co-IP and in vivo isoform-specific siRNA silencing for VAMP2; yeast two-hybrid, co-IP, pulse-chase, and glycosylation-mutagenesis for OS9

    PMID:25008321 PMID:26721884

    Open questions at the time
    • The E3 machinery cooperating with OS9 in NKCC2 ERAD not defined
  14. 2016 Medium

    Mapping inflammatory and hormonal modulators (NO/PDE5, TNF-alpha, IL-1R1, 20-HETE/Nedd4-2) positioned NKCC2 as an integration point for blood-pressure control.

    Evidence Native TAL activity assays with PDE5/NO pharmacology, knockout mice, and co-IP for ubiquitination/Nedd4-2

    PMID:12837683 PMID:21511694 PMID:23104236 PMID:23123217 PMID:26712462 PMID:26887831

    Open questions at the time
    • Many modulators rest on single-lab studies
    • Convergence of these signals on common NKCC2 regulatory residues not resolved

Open questions

Synthesis pass · forward-looking unresolved questions
  • How chloride-sensing, multiple kinase inputs, and trafficking machinery are integrated structurally and the discrepancy between WNK3's strong oocyte effect and minor in vivo role remain unresolved.
  • No structural model of NKCC2
  • In vivo hierarchy of WNK1/WNK3/WNK4 on NKCC2 not settled
  • Integration of metabolic, inflammatory, and hormonal regulation unmapped

Mechanism profile

Synthesis pass · controlled-vocabulary classification · explore literature graph →
Molecular activity
GO:0005215 transporter activity 4 GO:0140104 molecular carrier activity 3 GO:0140299 molecular sensor activity 2
Localization
GO:0005886 plasma membrane 4 GO:0005783 endoplasmic reticulum 3 GO:0031410 cytoplasmic vesicle 3
Pathway
R-HSA-162582 Signal Transduction 4 R-HSA-5653656 Vesicle-mediated transport 4 R-HSA-382551 Transport of small molecules 3 R-HSA-392499 Metabolism of proteins 3
Partners
MALNEDD4LOS9OSR1SCAMP2SPAKVAMP2WNK3

Evidence

Reading pass · 35 per-paper findings extracted from the source corpus
Year Finding Method Journal Conf PMIDs
1996 Loss-of-function mutations (frameshift and non-conservative missense) in the NKCC2 gene (SLC12A1) are the molecular cause of Bartter's syndrome type I, demonstrating that NKCC2 is required for renal Na-K-2Cl reabsorption in the thick ascending limb. Genetic linkage analysis and mutation identification in Bartter's syndrome patients; co-segregation of NKCC2 mutations with disease Nature genetics High 8640224
1996 BSC-1/NKCC2 protein is localized exclusively to the apical membrane of medullary and cortical thick ascending limb (TAL) segments in rat kidney; protein abundance is regulated by chronic saline loading and furosemide infusion (furosemide causes upward molecular mass shift and apparent increased expression). Peptide-derived polyclonal antibody immunoblotting and immunoperoxidase immunohistochemistry in rat kidney fractions The American journal of physiology High 8853424
2003 Short-term vasopressin (desmopressin) stimulation of NKCC2 in mouse kidney involves both increased phosphorylation of regulatory threonines in the NKCC2 N-terminus and membrane translocation, with a 55% increase in NKCC2 molecules at the apical membrane; phosphorylated NKCC2 is restricted to the cell membrane compartment. In vivo vasopressin analogue administration; phosphospecific antibody immunofluorescence; morphometric electron microscopy of apical membrane The Journal of biological chemistry High 12732642
2005 WNK3 kinase is a potent positive regulator of NKCC2 transport activity; kinase-active WNK3 activates NKCC2 by increasing its expression at the plasma membrane and increasing phosphorylation at Thr-184 and Thr-189, whereas kinase-inactive WNK3 potently inhibits NKCC2. Co-expression studies in Xenopus laevis oocytes; plasma membrane expression assays; phosphorylation site analysis Proceedings of the National Academy of Sciences of the United States of America High 16275913
2008 Intracellular chloride depletion activates NKCC2 by promoting phosphorylation of three conserved N-terminal threonines (T96, T101, T111); this chloride-sensing mechanism requires both WNK3 (the chloride-sensitive kinase, positioned upstream) and SPAK kinase. WNK3 activates NKCC2 via its SPAK-binding motif, and catalytically inactive WNK3 completely blocks chloride-depletion-induced NKCC2 activation. Co-expression in Xenopus oocytes with KCC2 or hypotonic stress; phosphospecific mutagenesis of T96/T101/T111; kinase-inactive WNK3 mutant; deletion of WNK3 SPAK-binding motif Proceedings of the National Academy of Sciences of the United States of America High 18550832
2005 NKCC2 transport activity is stimulated by hypertonicity and regulated by three N-terminal threonines (T99, T104, T117) that together constitute the regulatory phospho-domain; all three residues are required for the full hypertonic response, but none is individually necessary or sufficient. Under isotonic/hypotonic conditions NKCC2 retains ~50% activity even without phosphorylation of this domain. Mutagenic analysis of individual and combined threonine residues expressed in Xenopus oocytes; selective N-terminal deletions; chimeric NKCC1/NKCC2 constructs American journal of physiology. Renal physiology High 16077079
2005 cAMP increases surface expression of NKCC2 in rat medullary thick ascending limbs via a VAMP-dependent vesicle trafficking mechanism; tetanus toxin (which inactivates VAMP-2 and -3) completely blocks cAMP-stimulated surface NKCC2 expression and Cl- absorption. VAMP-2 and VAMP-3 localize to the subapical space of TAL cells. Surface biotinylation of rat mTAL suspensions; tetanus toxin VAMP inhibition; confocal microscopy; isolated perfused mTAL Cl- absorption measurements American journal of physiology. Renal physiology High 16144963
2009 cAMP stimulates NKCC2 surface expression in TAL specifically via protein kinase A (PKA), not Epac; PKA stimulates exocytic insertion of NKCC2 into the apical membrane (3-fold increase in exocytic insertion), while constitutive exocytosis is PKA-independent. Surface biotinylation in rat TALs; selective PKA agonist (N6-benzoyl-cAMP) vs. Epac agonist; H-89 PKA inhibitor; FM1-43 apical exocytosis assay; confocal imaging of isolated perfused TALs The Journal of biological chemistry High 19592485
2009 A trihydrophobic LLV motif (residues 1081-1083) in the distal C-terminus of NKCC2 is required for ER exit and cell surface expression; naturally occurring mutations depriving NKCC2 of this region cause ER retention, prevent complex glycosylation, and abolish surface delivery without affecting synthesis or degradation rates. Confocal microscopy; surface biotinylation; pulse-chase analysis; co-immunolocalization with ER marker PDI; proteasome/lysosome inhibitor treatment; serial truncation and site-directed mutagenesis The Journal of biological chemistry High 19535327
2011 SPAK and OSR1 kinases, activated by WNK1, interact with an RFQV motif on NKCC2 and directly phosphorylate Thr95, Thr100, Thr105 (and possibly Ser91) under hypotonic low-chloride conditions; a SPAK/OSR1-independent kinase (possibly AMPK) phosphorylates Ser130; Thr105 and Ser130 phosphorylation plays the most important role in stimulating NKCC2 activity. Unlike NCC, NKCC2 membrane translocation is not triggered by SPAK/OSR1 phosphorylation (NKCC2 is constitutively at the membrane). Phosphorylation site mapping by mass spectrometry and phosphospecific antibodies; RFQV motif mutation; kinase activity assays; expression of NKCC2 isoforms A, B, F in cells Journal of cell science High 21321328
2007 AMPK directly phosphorylates NKCC2 on Ser126 in vitro; AMPK physically associates with the N-terminal cytoplasmic domain of NKCC2 (co-precipitation); AMPK activation in MMDD1 cells increases Ser126 phosphorylation in situ; Ser126Ala mutation markedly reduces cotransporter activity under isotonic (basal) but not hypertonic conditions. In vitro kinase assay; co-precipitation; cell-based phosphorylation; functional Xenopus oocyte expression with S126A mutant The Biochemical journal High 17341212
2003 Six Bartter syndrome type I missense/frameshift mutations (G193R, A267S, G319R, A508T, del526N, Y998X) in human NKCC2 consistently abolish transport activity when expressed in Xenopus oocytes; mutant proteins show reduced expression, are routed to the plasma membrane, but are functionally impaired. Bumetanide-sensitive 22Na+ uptake assay in Xenopus oocytes; immunoblotting; immunocytochemistry Journal of the American Society of Nephrology : JASN High 12761241
2007 Aldolase B interacts with NKCC2 C-terminal tail (identified by yeast two-hybrid); co-immunoprecipitation and co-localization confirmed in renal cells; aldolase B co-expression reduces NKCC2 surface expression and transport activity; the substrate fructose 1,6-bisphosphate disrupts aldolase B binding and abolishes its effect on NKCC2 surface levels. Yeast two-hybrid screen; co-immunoprecipitation; co-immunolocalization; surface biotinylation; functional transport assay The Journal of biological chemistry High 17848580
2010 Dynamin-2 and clathrin (via clathrin-mediated endocytosis) and lipid rafts (including caveolin-1) mediate NKCC2 endocytosis at the apical surface in native TALs; simultaneous inhibition of clathrin- and lipid raft-mediated endocytosis completely blocks NKCC2 internalization. Blocking endocytosis increases steady-state surface NKCC2. Dynasore treatment; dominant-negative Dyn2K44A expression; chlorpromazine clathrin inhibition; synaptojanin-clathrin interaction blockade; methyl-β-cyclodextrin lipid raft disruption; caveolin-1 siRNA silencing; surface biotinylation in isolated rat THALs The Journal of biological chemistry High 22977238
2010 NKCC2 undergoes constitutive endocytosis (21.5% of surface pool per 30 min) and recycling (36% of retrieved NKCC2 returns to plasma membrane) in rat THALs; blockade of endocytosis with methyl-β-cyclodextrin (cholesterol chelation) increases steady-state surface NKCC2 by 60% and enhances NaCl entry by 57%. Surface biotinylation; Western blot; confocal microscopy of isolated perfused rat THALs; methyl-β-cyclodextrin treatment American journal of physiology. Renal physiology High 20719977
2014 VAMP2 (but not VAMP3) selectively mediates cAMP/PKA-stimulated NKCC2 exocytic delivery and surface expression in TALs; NKCC2 co-immunoprecipitates with VAMP2 in native rat TALs; cAMP stimulation enhances VAMP2 exocytosis and promotes VAMP2-NKCC2 co-immunoprecipitation; in vivo VAMP2 silencing completely blocks cAMP-stimulated NKCC2 exocytosis. VAMP2 is not involved in constitutive NKCC2 delivery. Co-immunoprecipitation in rat TALs; VAMP2/VAMP3 in vivo siRNA silencing; surface biotinylation; exocytosis assay The Journal of biological chemistry High 25008321
2011 Secretory carrier membrane protein 2 (SCAMP2) interacts with the NKCC2 C-terminus (yeast two-hybrid and co-IP); co-expression of SCAMP2 decreases NKCC2 surface expression and transport activity by impairing exocytotic trafficking (not endocytosis); SCAMP2 co-localizes with intracellularly retained NKCC2 in recycling endosomes; a single point mutation (C201A) in SCAMP2's E peptide abolishes its inhibitory effect. Yeast two-hybrid; co-immunoprecipitation; co-immunolocalization; surface biotinylation; MESNA cleavage endocytosis assay; E-peptide mutagenesis The Journal of biological chemistry High 21205824
2010 MAL/VIP17 co-localizes and co-immunoprecipitates with NKCC2 in LLC-PK1 cells and rat kidney medulla; a 150-amino acid stretch of the NKCC2 C-terminal tail mediates the interaction; MAL/VIP17 increases cell surface retention of NKCC2 by attenuating its internalization and coincides with increased NKCC2 phosphorylation. Transgenic overexpression of MAL/VIP17 in mouse kidney produces highly glycosylated and phosphorylated NKCC2. Co-immunoprecipitation; co-immunolocalization; surface retention assay; transgenic mouse overexpression; co-deletion mapping of interaction domain Molecular biology of the cell Medium 20861303
2015 OS9 protein interacts specifically with the immature (ER-localized) form of NKCC2 (identified by yeast two-hybrid and confirmed by co-IP); OS9 overexpression increases NKCC2 proteasomal degradation; OS9 knockdown by siRNA increases NKCC2 stability. OS9-induced degradation is N-glycan-dependent (NKCC2 N-glycosylation site mutations abolish OS9's effect) but MRH-domain-independent, defining an ERAD pathway specific to immature NKCC2. Yeast two-hybrid; co-immunoprecipitation; immunocytochemistry; pulse-chase and cycloheximide-chase; siRNA knockdown; proteasome inhibitor MG132; N-glycosylation and MRH-domain mutagenesis The Journal of biological chemistry High 26721884
2012 Multiple evolutionarily conserved di-leucine-like motifs in the NKCC2 C-terminus (1038LL1039, 1048LI1049, and 1081LLV1083) are each required for ER exit and cell surface expression; double mutation of any one pair to di-alanine disrupts glycosylation and surface expression by causing ER retention, without affecting synthesis or degradation rates. Serial C-terminal truncations; site-directed mutagenesis; pulse-chase analysis; co-immunolocalization with ER marker calnexin; surface biotinylation; multiple expression systems The Journal of biological chemistry High 23105100
2006 The ion affinity differences among NKCC2 splice variants (F, A, B) are determined by specific residues in the second transmembrane domain (TM2) and the putative intracellular loop (ICL1) connecting TM2 and TM3; six residue substitutions convert the B variant into the F variant; involvement of ICL1 residues suggests this region may be membrane-embedded and contribute to chloride binding. Site-directed mutagenesis of individual and combined residues in NKCC2B; functional expression in Xenopus oocytes with ion affinity measurements The Journal of biological chemistry High 17186942
1998 NKCC2 (rabbit NKCC2A chimera) expressed in HEK-293 cells has a 4-fold lower Rb+ affinity and 3-fold higher bumetanide affinity compared to NKCC1; NKCC2 activity is increased by low-[Cl-] media; NKCC2 exhibits appropriate volume response unlike NKCC1, supporting a model where apical NKCC2 activity is matched to basolateral Cl- exit via changes in intracellular [Cl-]. Stable expression of rabbit NKCC2A chimera in HEK-293 cells; ion affinity kinetic measurements; low-[Cl-] activation assays; volume response assay The Journal of biological chemistry High 9556622
2011 Tamm-Horsfall protein (THP) facilitates NKCC2 activation in a chloride-sensitive manner; THP-deficient mice show increased intracellular NKCC2 in subapical vesicles and decreased basal NKCC2 phosphorylation (-49%); THP co-expression in oocytes enhances NKCC2 activation under low-chloride hypotonic stress; vasopressin-stimulated NKCC2 phosphorylation is blunted in THP absence. THP knockout mice; immunofluorescence; surface biotinylation; Xenopus oocyte co-injection; cultured TAL cells with THP transfection; V2 receptor agonist stimulation The Journal of biological chemistry High 21737451
2010 Kidney-specific (KS)-WNK1 is a negative regulator of NKCC2 in vivo; transgenic KS-WNK1 overexpression reduces surface expression of total and phosphorylated NKCC2 in thick ascending limb; targeted deletion of exon 4A (KS-WNK1-specific exon) increases surface expression of total and phosphorylated NKCC2. Transgenic mouse overexpression and knockout of KS-WNK1; immunofluorescent staining of total and phosphorylated NKCC2 in kidney sections Human molecular genetics High 21131289
2012 NKCC2 does not cotransport water, in contrast to NKCC1 which cotransports ~460-600 water molecules per turnover; osmotic gradients did not induce water transport in NKCC2-expressing oocytes, whereas NKCC1 supports bumetanide-blockable, uphill water transport. Expression of NKCC1 and NKCC2 in Xenopus oocytes; volume measurements; 86Rb+ ion flux assays; bumetanide inhibition The Journal of physiology High 22250214
2011 TNF-alpha acts as an endogenous inhibitor of NKCC2 isoform A expression and activity in thick ascending limbs; TNF gene deletion doubles total NKCC2 protein, increases NKCC2A mRNA 4-fold, and increases bumetanide-sensitive O2 consumption (a correlate of NKCC2 activity) by 2-fold; hTNF replacement restores NKCC2 expression and activity. TNF knockout mice; Western blotting; RT-PCR; bumetanide-sensitive O2 consumption in isolated mTAL tubules; hTNF rescue experiment American journal of physiology. Renal physiology Medium 21511694
2012 Adenylyl cyclase 6 (AC6) mediates vasopressin-induced phosphorylation of NKCC2 at S126 and determines total NKCC2 protein abundance in the medullary TAL; AC6 knockout mice lack desmopressin-stimulated S126 NKCC2 phosphorylation and have lower NKCC2 expression with a mild Bartter syndrome-like phenotype. AC6 knockout mice; desmopressin stimulation; phosphospecific Western blotting for pS126 NKCC2; immunohistochemistry The American journal of pathology Medium 23123217
2015 IL-1 receptor (IL-1R1) activation potentiates sodium reabsorption via NKCC2 in the nephron; IL-1R1 deficiency or blockade reduces blood pressure by mitigating NKCC2-dependent sodium reabsorption; the mechanism involves IL-1R1 preventing intra-renal myeloid cells from maturing into Ly6C+Ly6G- macrophages that suppress NKCC2 activity via nitric oxide. IL-1R1 knockout mice; angiotensin II-induced hypertension model; diuretic response assays; macrophage characterization by flow cytometry Cell metabolism Medium 26712462
2015 NKCC2 is expressed in the hypothalamo-neurohypophyseal system (HNS) of the brain, not only the kidney; HNS NKCC2 is upregulated by osmotic stress; knockdown of HNS NKCC2 impairs fluid balance after high-salt ingestion; dehydration-evoked GABA-mediated excitation of AVP neurons is reversed by bumetanide, and furosemide blocks AVP release in vivo and in hypothalamic explants. In situ hybridization; RT-PCR; shRNA knockdown of HNS NKCC2 in rats; bumetanide/furosemide pharmacology in vivo and in hypothalamic explants; electrophysiology of AVP neurons The Journal of neuroscience : the official journal of the Society for Neuroscience Medium 25834041
2011 Rare human NKCC2 mutations associated with lower blood pressure exhibit impaired protein processing (reduced complex glycosylation, absence of plasma membrane localization for R302W and L505V) or reduced transport function; P569H mutation reduces sodium affinity by 50%; P254A increases rubidium affinity by 35%; functional variants retain regulation by cell volume and intracellular chloride. Heterologous expression in Xenopus oocytes and HEK-293 cells; 86Rb+ transport assay; surface membrane localization; glycosylation analysis; ion affinity kinetics American journal of physiology. Renal physiology High 21209010
2011 WNK3 knockout mice show no significant decrease in NKCC2 phosphorylation or expression under normal or low-salt diet, indicating that WNK3 plays only a minor role in regulating NKCC2 phosphorylation in vivo, with compensation by WNK4 and WNK1. WNK3 knockout mice; Western blotting for phospho-NKCC2, phospho-OSR1, phospho-SPAK; urine Na+/K+ excretion; blood pressure telemetry Biology open Medium 23213404
2003 NO positively regulates NKCC2 protein abundance in kidney; NO synthase inhibition with L-NAME in aldosterone-treated rats decreases NKCC2 protein abundance without changes in corresponding mRNA levels, indicating post-translational regulation of NKCC2 by NO. In vivo L-NAME infusion in aldosterone-escape model; semiquantitative immunoblotting; mRNA measurements American journal of physiology. Renal physiology Medium 12837683
2012 20-HETE and high salt synergistically decrease NKCC2 protein expression via Nedd4-2-mediated ubiquitin-proteasome degradation; NKCC2 was found to be ubiquitinated and to interact with Nedd4-2 in transgenic CYP4F2 mice on high-salt diet; proteasome inhibition or inhibition of 20-HETE synthesis restores NKCC2 expression. Immunoprecipitation for ubiquitin and Nedd4-2 interaction with NKCC2; proteasome inhibitor rescue; Western blotting in CYP4F2 transgenic and WT mice on high-salt diet Human genetics Medium 23104236
2016 Angiotensin II-induced hypertension impairs NO-mediated inhibition of NKCC2 activity in TALs via enhanced PDE5-mediated cGMP degradation; a PDE5 inhibitor (vardenafil) restores NO's ability to inhibit NKCC2 and increase cGMP; dibutyryl-cGMP reduces NKCC2 activity equally in vehicle and ANG II hypertensive rats, placing the defect upstream of cGMP action. Isolated perfused rat THAL NKCC2 activity assay; NO donor and ET-1 stimulation; dibutyryl-cGMP; PDE5 inhibitor vardenafil; cGMP measurement; ANG II infusion model American journal of physiology. Renal physiology Medium 26887831
2008 The three NKCC2 splice isoforms (B, A, F) differ in ion affinities and show distinct localization along the TAL; isoform-specific NKCC2 knockout studies demonstrate that NKCC2B and NKCC2A cooperate in macula densa cells to facilitate efficient salt sensing over wide ranges of salt concentrations. Differential splicing analysis; RT-PCR; isoform-specific NKCC2 knockout mice; localization studies along the TAL American journal of physiology. Renal physiology Medium 18495801

Source papers

Stage 0 corpus · 100 papers · ranked by NIH iCite citations
Year Title Journal Citations PMID
1984 Growth inhibitor from BSC-1 cells closely related to platelet type beta transforming growth factor. Science (New York, N.Y.) 778 6093254
1996 Bartter's syndrome, hypokalaemic alkalosis with hypercalciuria, is caused by mutations in the Na-K-2Cl cotransporter NKCC2. Nature genetics 713 8640224
2000 Gemcitabine plus best supportive care (BSC) vs BSC in inoperable non-small cell lung cancer--a randomized trial with quality of life as the primary outcome. UK NSCLC Gemcitabine Group. Non-Small Cell Lung Cancer. British journal of cancer 263 10945489
1975 Methylated simian virus 40-specific RNA from nuclei and cytoplasm of infected BSC-1 cells. Proceedings of the National Academy of Sciences of the United States of America 208 166375
2008 Regulation of NKCC2 by a chloride-sensing mechanism involving the WNK3 and SPAK kinases. Proceedings of the National Academy of Sciences of the United States of America 195 18550832
1996 Localization and regulation of the rat renal Na(+)-K(+)-2Cl- cotransporter, BSC-1. The American journal of physiology 184 8853424
2006 The Atx1-Ccc2 complex is a metal-mediated protein-protein interaction. Nature chemical biology 181 16732294
2003 Short-term stimulation of the renal Na-K-Cl cotransporter (NKCC2) by vasopressin involves phosphorylation and membrane translocation of the protein. The Journal of biological chemistry 173 12732642
2000 Energetics of copper trafficking between the Atx1 metallochaperone and the intracellular copper transporter, Ccc2. The Journal of biological chemistry 173 10764731
1991 Microsurgical removal of centrosomes blocks cell reproduction and centriole generation in BSC-1 cells. Cell 173 1934057
2011 Regulation of the NKCC2 ion cotransporter by SPAK-OSR1-dependent and -independent pathways. Journal of cell science 162 21321328
1977 Density-dependent regulation of growth of BSC-1 cells in cell culture: control of growth by serum factors. Proceedings of the National Academy of Sciences of the United States of America 153 303774
2005 WNK3 kinase is a positive regulator of NKCC2 and NCC, renal cation-Cl- cotransporters required for normal blood pressure homeostasis. Proceedings of the National Academy of Sciences of the United States of America 152 16275913
2002 Upregulation of renal BSC1 and TSC in prenatally programmed hypertension. American journal of physiology. Renal physiology 148 12060603
2011 Molecular regulation of NKCC2 in the thick ascending limb. American journal of physiology. Renal physiology 145 21900458
2011 Activation of the bumetanide-sensitive Na+,K+,2Cl- cotransporter (NKCC2) is facilitated by Tamm-Horsfall protein in a chloride-sensitive manner. The Journal of biological chemistry 143 21737451
1988 Amino acid sequence of the BSC-1 cell growth inhibitor (polyergin) deduced from the nucleotide sequence of the cDNA. Proceedings of the National Academy of Sciences of the United States of America 142 3277172
1995 Sequence, mapping and disruption of CCC2, a gene that cross-complements the Ca(2+)-sensitive phenotype of csg1 mutants and encodes a P-type ATPase belonging to the Cu(2+)-ATPase subfamily. Yeast (Chichester, England) 136 7785328
1985 BSC-1 growth inhibitor transforms a mitogenic stimulus into a hypertrophic stimulus for renal proximal tubular cells: relationship to Na+/H+ antiport activity. Proceedings of the National Academy of Sciences of the United States of America 129 2994063
2015 Interleukin-1 Receptor Activation Potentiates Salt Reabsorption in Angiotensin II-Induced Hypertension via the NKCC2 Co-transporter in the Nephron. Cell metabolism 124 26712462
1999 Relationship between quality of life and clinical outcomes in advanced non-small cell lung cancer: best supportive care (BSC) versus BSC plus chemotherapy. Lung cancer (Amsterdam, Netherlands) 114 10403690
1978 Density-dependent regulation of growth of BSC-1 cells in cell culture: growth inhibitors formed by the cells. Proceedings of the National Academy of Sciences of the United States of America 111 273914
1998 Comparison of Na-K-Cl cotransporters. NKCC1, NKCC2, and the HEK cell Na-L-Cl cotransporter. The Journal of biological chemistry 110 9556622
2005 Regulatory phosphorylation sites in the NH2 terminus of the renal Na-K-Cl cotransporter (NKCC2). American journal of physiology. Renal physiology 103 16077079
2013 Physiology and pathophysiology of SLC12A1/2 transporters. Pflugers Archiv : European journal of physiology 102 24097229
2002 Time course of renal Na-K-ATPase, NHE3, NKCC2, NCC, and ENaC abundance changes with dietary NaCl restriction. American journal of physiology. Renal physiology 100 12217855
2005 cAMP increases surface expression of NKCC2 in rat thick ascending limbs: role of VAMP. American journal of physiology. Renal physiology 96 16144963
2008 Thick ascending limb: the Na(+):K (+):2Cl (-) co-transporter, NKCC2, and the calcium-sensing receptor, CaSR. Pflugers Archiv : European journal of physiology 95 18982348
2002 Rat NKCC2/NKCC1 cotransporter selectivity for loop diuretic drugs. Naunyn-Schmiedeberg's archives of pharmacology 90 11882915
2010 Downregulation of NCC and NKCC2 cotransporters by kidney-specific WNK1 revealed by gene disruption and transgenic mouse models. Human molecular genetics 85 21131289
2012 Cotransport of water by Na⁺-K⁺-2Cl⁻ cotransporters expressed in Xenopus oocytes: NKCC1 versus NKCC2. The Journal of physiology 77 22250214
2007 Regulation of the renal-specific Na+-K+-2Cl- co-transporter NKCC2 by AMP-activated protein kinase (AMPK). The Biochemical journal 77 17341212
2009 cAMP stimulates apical exocytosis of the renal Na(+)-K(+)-2Cl(-) cotransporter NKCC2 in the thick ascending limb: role of protein kinase A. The Journal of biological chemistry 73 19592485
1992 Mechanism of centrosome positioning during the wound response in BSC-1 cells. The Journal of cell biology 71 1740470
2008 Isoforms of renal Na-K-2Cl cotransporter NKCC2: expression and functional significance. American journal of physiology. Renal physiology 70 18495801
1974 Vaccinia virus replication in enucleate BSC-1 cells: particle production and synthesis of viral DNA and proteins. Journal of virology 69 4204192
2012 Adenylyl cyclase 6 enhances NKCC2 expression and mediates vasopressin-induced phosphorylation of NKCC2 and NCC. The American journal of pathology 65 23123217
1999 Altered expression of Na transporters NHE-3, NaPi-II, Na-K-ATPase, BSC-1, and TSC in CRF rat kidneys. The American journal of physiology 64 10444581
1982 Serum and epidermal growth factor transiently depolarize quiescent BSC-1 epithelial cells. Proceedings of the National Academy of Sciences of the United States of America 63 6984193
2002 Altered expression of renal NHE3, TSC, BSC-1, and ENaC subunits in potassium-depleted rats. American journal of physiology. Renal physiology 62 12388387
1992 Mutations within the 5' nontranslated region of hepatitis A virus RNA which enhance replication in BS-C-1 cells. Journal of virology 59 1404601
1987 Diphtheria toxin receptor. Identification of specific diphtheria toxin-binding proteins on the surface of Vero and BS-C-1 cells. The Journal of biological chemistry 52 3654609
2004 Elevated BSC-1 and ROMK expression in Dahl salt-sensitive rat kidneys. Hypertension (Dallas, Tex. : 1979) 50 14967839
2015 NKCC1 and NKCC2: The pathogenetic role of cation-chloride cotransporters in hypertension. Genes & diseases 48 26114157
2011 Rare mutations in SLC12A1 and SLC12A3 protect against hypertension by reducing the activity of renal salt cotransporters. Journal of hypertension 48 21157372
2003 Mutations in the human Na-K-2Cl cotransporter (NKCC2) identified in Bartter syndrome type I consistently result in nonfunctional transporters. Journal of the American Society of Nephrology : JASN 48 12761241
2002 Renal Na-K-Cl cotransporter NKCC2 in Dahl salt-sensitive rats. Journal of hypertension 48 11910309
2011 High salt differentially regulates surface NKCC2 expression in thick ascending limbs of Dahl salt-sensitive and salt-resistant rats. American journal of physiology. Renal physiology 47 21307126
2011 A minor role of WNK3 in regulating phosphorylation of renal NKCC2 and NCC co-transporters in vivo. Biology open 45 23213404
2009 A highly conserved motif at the COOH terminus dictates endoplasmic reticulum exit and cell surface expression of NKCC2. The Journal of biological chemistry 45 19535327
2003 Regulation of NHE3, NKCC2, and NCC abundance in kidney during aldosterone escape phenomenon: role of NO. American journal of physiology. Renal physiology 45 12837683
2012 Dynamin2, clathrin, and lipid rafts mediate endocytosis of the apical Na/K/2Cl cotransporter NKCC2 in thick ascending limbs. The Journal of biological chemistry 44 22977238
2001 Regulation of the sodium transporters NHE3, NKCC2 and NCC in the kidney. Current opinion in nephrology and hypertension 44 11496061
2012 Effect of heterozygous deletion of WNK1 on the WNK-OSR1/ SPAK-NCC/NKCC1/NKCC2 signal cascade in the kidney and blood vessels. Clinical and experimental nephrology 43 22294159
1997 Caenorhabditis elegans cDNA for a Menkes/Wilson disease gene homologue and its function in a yeast CCC2 gene deletion mutant. Journal of biochemistry 43 9354393
2011 Rare mutations in the human Na-K-Cl cotransporter (NKCC2) associated with lower blood pressure exhibit impaired processing and transport function. American journal of physiology. Renal physiology 42 21209010
2018 Pre-eclampsia is associated with altered expression of the renal sodium transporters NKCC2, NCC and ENaC in urinary extracellular vesicles. PloS one 40 30248150
2011 Tumor necrosis factor-alpha is an endogenous inhibitor of Na+-K+-2Cl- cotransporter (NKCC2) isoform A in the thick ascending limb. American journal of physiology. Renal physiology 40 21511694
1997 A common NKCC2 mutation in Costa Rican Bartter's syndrome patients: evidence for a founder effect. Journal of the American Society of Nephrology : JASN 40 9355073
2008 Treating lithium-induced nephrogenic diabetes insipidus with a COX-2 inhibitor improves polyuria via upregulation of AQP2 and NKCC2. American journal of physiology. Renal physiology 39 18216147
2006 Acute growth hormone administration induces antidiuretic and antinatriuretic effects and increases phosphorylation of NKCC2. American journal of physiology. Renal physiology 39 17062845
2004 Increased expression of the sodium transporter BSC-1 in spontaneously hypertensive rats. The Journal of pharmacology and experimental therapeutics 39 15340004
2009 In vivo and in vitro splicing assay of SLC12A1 in an antenatal salt-losing tubulopathy patient with an intronic mutation. Human genetics 38 19513753
2006 The residues determining differences in ion affinities among the alternative splice variants F, A, and B of the mammalian renal Na-K-Cl cotransporter (NKCC2). The Journal of biological chemistry 38 17186942
2013 Dietary salt intake modulates differential splicing of the Na-K-2Cl cotransporter NKCC2. American journal of physiology. Renal physiology 37 23946287
2004 Reduced renal Na+-K+-Cl- co-transporter activity and inhibited NKCC2 mRNA expression by Leptospira shermani: from bed-side to bench. Nephrology, dialysis, transplantation : official publication of the European Dialysis and Transplant Association - European Renal Association 37 15388818
2008 Expression of aquaporin1, 3, and 4, NKCC1, and NKCC2 in the human endolymphatic sac. Auris, nasus, larynx 36 18606512
1983 Rapid selective effects by a growth inhibitor and epidermal growth factor on the incorporation of [35S]methionine into proteins secreted by African green monkey (BSC-1) cells. Proceedings of the National Academy of Sciences of the United States of America 36 6604275
2007 NKCC2 surface expression in mammalian cells: down-regulation by novel interaction with aldolase B. The Journal of biological chemistry 35 17848580
2015 Osmoregulation requires brain expression of the renal Na-K-2Cl cotransporter NKCC2. The Journal of neuroscience : the official journal of the Society for Neuroscience 34 25834041
2010 Localization and functional characterization of the human NKCC2 isoforms. Acta physiologica (Oxford, England) 34 20146722
2010 Mutation of the Na(+)-K(+)-2Cl(-) cotransporter NKCC2 in mice is associated with severe polyuria and a urea-selective concentrating defect without hyperreninemia. American journal of physiology. Renal physiology 34 20219826
2003 Differential expression pattern of chloride transporters NCC, NKCC2, KCC1, KCC3, KCC4, and AE3 in the developing rat auditory brainstem. Cell and tissue research 34 12712325
1993 Synthesis and assembly of hepatitis A virus-specific proteins in BS-C-1 cells. Journal of virology 34 8388489
1978 Density-dependent regulation of growth of BSC-1 cells in cell culture: control of growth by low molecular weight nutrients. Proceedings of the National Academy of Sciences of the United States of America 34 272650
2010 Constitutive endocytosis and recycling of NKCC2 in rat thick ascending limbs. American journal of physiology. Renal physiology 32 20719977
2007 Chronic noradrenaline increases renal expression of NHE-3, NBC-1, BSC-1 and aquaporin-2. Clinical and experimental pharmacology & physiology 32 18177483
2006 Rosiglitazone regulates ENaC and Na-K-2Cl cotransporter (NKCC2) abundance in the obese Zucker rat. American journal of nephrology 31 16757903
2016 Spilanthol from Acmella Oleracea Lowers the Intracellular Levels of cAMP Impairing NKCC2 Phosphorylation and Water Channel AQP2 Membrane Expression in Mouse Kidney. PloS one 30 27213818
2015 OS9 Protein Interacts with Na-K-2Cl Co-transporter (NKCC2) and Targets Its Immature Form for the Endoplasmic Reticulum-associated Degradation Pathway. The Journal of biological chemistry 30 26721884
2014 Vesicle-associated membrane protein 2 (VAMP2) but Not VAMP3 mediates cAMP-stimulated trafficking of the renal Na+-K+-2Cl- co-transporter NKCC2 in thick ascending limbs. The Journal of biological chemistry 30 25008321
2008 Chronic candesartan alters expression and activity of NKCC2, NCC, and ENaC in the obese Zucker rat. American journal of physiology. Renal physiology 30 18305093
2014 Effects of NKCC2 isoform regulation on NaCl transport in thick ascending limb and macula densa: a modeling study. American journal of physiology. Renal physiology 29 24848496
2011 Secretory carrier membrane protein 2 regulates exocytic insertion of NKCC2 into the cell membrane. The Journal of biological chemistry 29 21205824
2010 MAL/VIP17, a new player in the regulation of NKCC2 in the kidney. Molecular biology of the cell 29 20861303
2014 Novel mechanisms of Na+ retention in obesity: phosphorylation of NKCC2 and regulation of SPAK/OSR1 by AMPK. American journal of physiology. Renal physiology 27 24808538
2012 Multiple evolutionarily conserved Di-leucine like motifs in the carboxyl terminus control the anterograde trafficking of NKCC2. The Journal of biological chemistry 26 23105100
2006 Sequential expression of NKCC2, TonEBP, aldose reductase, and urea transporter-A in developing mouse kidney. American journal of physiology. Renal physiology 26 16926446
2012 Synergistical effect of 20-HETE and high salt on NKCC2 protein and blood pressure via ubiquitin-proteasome pathway. Human genetics 25 23104236
2009 Expression of NKCC2 in the rat gastrointestinal tract. Neurogastroenterology and motility 25 19460103
2016 Angiotensin II-mediated hypertension impairs nitric oxide-induced NKCC2 inhibition in thick ascending limbs. American journal of physiology. Renal physiology 24 26887831
2012 Expression of the Slc12a1 gene in pancreatic β-cells: molecular characterization and in silico analysis. Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology 24 22759959
2011 Differential regulation of NFAT5 by NKCC2 isoforms in medullary thick ascending limb (mTAL) cells. American journal of physiology. Renal physiology 24 21228109
2011 NKCC2 is activated in Milan hypertensive rats contributing to the maintenance of salt-sensitive hypertension. Pflugers Archiv : European journal of physiology 24 21553016
2010 Differential regulation of the bumetanide-sensitive cotransporter (NKCC2) by ovarian hormones. Steroids 24 20580730
2007 Novel SLC12A1 (NKCC2) mutations in two families with Bartter syndrome type 1. Endocrine journal 24 17998760
2001 Alternative splicing of the BSC1 gene generates tissue-specific isoforms in the German cockroach. Insect biochemistry and molecular biology 24 11267908
1996 Inhibition of initiation of simian virus 40 DNA replication in infected BSC-1 cells by the DNA alkylating drug adozelesin. The Journal of biological chemistry 23 8702695
1995 Effects of the DNA-damaging enediyne C-1027 on intracellular SV40 and genomic DNA in green monkey kidney BSC-1 cells. Biochemistry 23 7849041
2010 Integrated genomic profiling identifies candidate genes implicated in glioma-genesis and a novel LEO1-SLC12A1 fusion gene. Genes, chromosomes & cancer 22 20196086

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