ライフサイエンスコーパス: NKCC2

1 n activated Na(+)-K(+)-2Cl(-) cotransporter (NKCC2).

2 s (WNKs)] as well as its substrates (NCC and NKCC2).

3 the apical Na(+)/K(+)/2Cl(-) co-transporter NKCC2.

4 chloride affinity and transport activity of NKCC2.

5 ot sufficient for anterograde trafficking of NKCC2.

6 domain of OS9 had no effect on its action on NKCC2.

7 osmotic stimulatory effect was observed with NKCC2.

8 ted protein (MAL)/VIP17 in the regulation of NKCC2.

9 ated chimeras of the mouse NKCC1 and the rat NKCC2.

10 ded by an additional exon in NKCC1 absent in NKCC2.

11 sodium-potassium-chloride co-transporter 2, NKCC2.

12 ion-chloride cotransporters KCC3, NKCC1, and NKCC2.

13 gulatory threonines in the amino terminus of NKCC2.

14 olved in calcineurin-dependent modulation of NKCC2.

15 to mutations in the Na-K-2Cl co-transporter, NKCC2.

16 e key components in the phosphoregulation of NKCC2.

17 ) as a novel and specific binding partner of NKCC2.

18 ciation involves mainly the immature form of NKCC2.

19 in partners specifically involved in ERAD of NKCC2.

20 is known about phosphatases that deactivate NKCC2.

21 protein degradation of the immature form of NKCC2.

22 phorylation as well as expression of NCC and NKCC2.

23 munoprecipitation of SPAK with WNK1, NCC and NKCC2.

24 ignaling via PKA and other kinases activates NKCC2.

25 toward the Na(+)-K(+)-2Cl(-) cotransporter, NKCC2.

26 the apical Na(+)/K(+)/2Cl(-) co-transporter NKCC2.

27 ase A (PKA), increasing steady-state surface NKCC2.

28 ocytosis and promotes VAMP2 interaction with NKCC2.

29 ical translocation and surface expression of NKCC2.

30 e is known about partners that interact with NKCC2.

31 QP2 (121+/-4 versus 100+/-1%; P<0.001), ISOM NKCC2 (133+/-1 versus 100+/-4%; P<0.05), and cortex plus

32 nd cortex plus outer stripe of outer medulla NKCC2 (142+/-16 versus 100+/-9%; P<0.05).

33 Mutations in the Na-K-2Cl cotransporter (NKCC2), a mediator of renal salt reabsorption, cause Bar

34 pite a lack of SPAK and OSR1, phosphorylated NKCC2 abundance was still high, suggesting the existence

35 there was no Bartter phenotype with reduced NKCC2 activity and increased NCC expression in Romk1(-/-

36 ted NOS3 and Akt by Western blotting, and 4) NKCC2 activity by fluorescence microscopy.

37 ion of NKCC2 at S126 might help to stabilize NKCC2 activity in the absence of AC6.

38 hese results support a model in which apical NKCC2 activity is matched to basolateral Cl exit through

39 NKCC2 activity is modulated by N-terminal phosphorylatio

40 Endothelin-1 inhibited NKCC2 activity, an effect that was blocked by dominant-n

41 NKCC2 and AnxA2 interact in a phosphorylation-dependent

42 We have previously shown that NKCC2 and its disease-causing mutants are subject to reg

43 ve modulation of ERAD components specific to NKCC2 and its disease-causing mutants might provide nove

44 By mapping NKCC2 and KCC2 antibody staining on these dendrites, we

45 ing on these dendrites, we further show that NKCC2 and KCC2 are preferentially located in the proxima

46 depolarization and hyperpolarization at the NKCC2 and KCC2 compartments, respectively, and underlies

47 We show here that selective blockade of the NKCC2 and KCC2 cotransporters located on starburst dendr

48 We demonstrated that 1) NKCC2 and MAL/VIP17 colocalize and coimmunoprecipitate i

49 We conclude that VAMP3 interacts with NKCC2 and mediates its constitutive exocytic delivery to

50 nction of the sodium chloride cotransporters NKCC2 and NCC (key components of salt reabsorption in th

51 nactive state, WNK3 is a potent inhibitor of NKCC2 and NCC activity.

52 R1-independent phosphorylation sites on both NKCC2 and NCC and changes in sodium transport along the

53 phosphorylate the ion cotransporters NKCC1, NKCC2 and NCC, leading to the identification of several

54 se-active WNK3 is a potent activator of both NKCC2 and NCC-mediated transport.

55 ot in OSR1 to the same degree, and activated NKCC2 and NCC.

56 ly, immunocytochemistry analysis showed that NKCC2 and OS9 co-localize at the endoplasmic reticulum.

57 lipid raft-associated trafficking factor for NKCC2 and provides mechanistic insight into the regulati

58 Phosphorylation of S126 NKCC2 and T58 NCC, induced by the V2 receptor agonist (1

59 )]vasopressin increases steady-state surface NKCC2 and that the protein kinase A (PKA) inhibitor H-89

60 screening identified an interaction between NKCC2 and the cytosolic protein, annexin A2 (AnxA2).

61 Measurements of steady-state surface NKCC2 and the rate of NKCC2 endocytosis in freshly isola

62 e kinases, regulate the furosemide-sensitive NKCC2 and the thiazide-sensitive NCC, kidney-specific CC

63 f +/+, yet no differences were found between NKCC2 +/+ and +/- mice in BP, blood gas, electrolytes, c

64 tion of the Na(+)/K(+)/2Cl(-) cotransporter (NKCC2) and AQP2, with less phosphorylation of AQP2 at se

65 on of ion transporters in the loop of Henle (NKCC2) and distal nephron (NCC, ENaC, and pendrin) as we

66 tion in three genes-SLC12A3 (NCCT), SLC12A1 (NKCC2) and KCNJ1 (ROMK)-causing rare recessive diseases

67 nsporters Na(+) -K(+) -2Cl(-) cotransporter (NKCC2) and Na(+) -Cl(-) cotransporter (NCC) via phosphor

68 e renal Na(+) -K(+) -2Cl(-) cotransporter 2 (NKCC2) and Na(+) Cl(-) cotransporter (NCC).

69 sporter (NKCC2) at serine residue 126 (pS126 NKCC2) and of the Na-Cl cotransporter (NCC) at threonine

70 tion of the Na(+)-K(+)-2Cl(-)-cotransporter (NKCC2) and the Na(+)-Cl(-)-cotransporter (NCC) by vasopr

71 ium, two chloride co-transporters, NKCC1 and NKCC2, and also affect other related ion co-transporters

72 echanism associated with mutations depriving NKCC2, and also all other members of the SLC12A family,

73 composed of the levels of mRNA for vimentin, NKCC2, and E-cadherin and of 18S ribosomal RNA provided

74 m-transporting proteins, including NaPi-IIa, NKCC2, and ENaC, did not change, although the abundance

75 ary expression of the Na-K-2Cl-cotransporter NKCC2, and greater furosemide-sensitive Na+ reabsorption

76 ome to the renal Na-K-2Cl cotransporter gene NKCC2, and identify frameshift or non-conservative misse

77 e regulatory threonine residues among NKCC1, NKCC2, and NCC family members, together with the fact th

78 Cross linking studies of NHE3, NKCC2, and NCC revealed that high molecular weight compl

79 +)-dependent chloride cotransporters (NKCC1, NKCC2, and NCC) are activated by phosphorylation to play

80 PAK/OSR1 to bind to and phosphorylate NKCC1, NKCC2, and NCC.

81 MMDD1 cells express COX-2, bNOS, NKCC2, and ROMK, but not Tamm-Horsfall protein, and show

82 acids are highly conserved between NKCC1 and NKCC2, and similarities are also present in the Na-Cl co

83 we examine the kinetic properties of NKCC1, NKCC2, and the endogenous HEK-293 cell cotransporter.

84 fferences among the three splice variants of NKCC2, and they support a model in which a reentrant loo

85 um-dependent sodium-chloride co-transporter, NKCC2, and thiazide-sensitive sodium-chloride cotranspor

86 n in mice augmented the abundance of phospho-NKCC2, and treatment of isolated TAL with cyclosporine i

87 companied by a significant decrease in THP-, NKCC2- and AQP1-positive loop of Henle nephron segments

88 bumetanide-sensitive Na-K-2Cl cotransporter [NKCC2]), and the distal convoluted tubule (the thiazide-

89 due to loss-of-function mutations in NCC and NKCC2 are consistent, in part, with their functional rol

90 quired for ER exit and surface expression of NKCC2 are evolutionarily conserved in all members of the

91 the existence of three different isoforms of NKCC2 are unclear.

92 variants of the renal Na-K-Cl cotransporter (NKCC2) are found in distinct regions of the thick ascend

93 ransporter (NCC) and Na-K-2Cl cotransporter (NKCC2) are involved in Gitelman and Bartter syndrome, re

94 Here, we identified NKCC2 as a target of the calcineurin Abeta isoform.

95 asopressin also increased phosphorylation of NKCC2 at both Ser126 (more than fivefold) and Ser874 (mo

96 Increased AC6-independent phosphorylation of NKCC2 at S126 might help to stabilize NKCC2 activity in

97 la, there was significant phosphorylation of NKCC2 at SPAK/OSR1-dependent sites despite a complete ab

98 Phosphorylation of NKCC2 at SPAK/OSR1-dependent sites was lower than in SPA

99 P17 also is involved in the stabilization of NKCC2 at the apical membrane in vivo.

100 exocytic delivery, decreasing the amount of NKCC2 at the TAL apical surface.

101 osphorylation of the Na-K-2Cl cotransporter (NKCC2) at serine residue 126 (pS126 NKCC2) and of the Na

102 er874 of the Na(+):K(+):2Cl(-) cotransporter NKCC2, at Ser552 of the Na(+):H(+) exchanger NHE3, and a

103 es a selective loss of NCC function, whereas NKCC2 becomes hyperphosphorylated.

104 porin 2, or Na(+)-K(+)-2Cl(-) co-transporter NKCC2/BSC1 protein abundances or UT-A1 mRNA abundance in

105 ice isoforms of the Na-K-2Cl co-transporter (NKCC2/BSC1) are expressed along the thick ascending limb

106 R1) with the cotransporters KCC3, NKCC1, and NKCC2 but not KCC1 and KCC4.

107 38 +/- 8% and increased steady-state surface NKCC2 by 37 +/- 8%, without changing total NKCC2 express

108 d surface expression and raft association of NKCC2 by 5-fold upon low chloride hypotonic stimulation,

109 6 +/- 11% and increased steady-state surface NKCC2 by 67 +/- 27% (p < 0.05).

110 IP17 increases the cell surface retention of NKCC2 by attenuating its internalization, and 4) this co

111 cAMP stimulates NKCC2 by enhancing steady-state apical membrane levels o

112 cAMP stimulates steady-state apical surface NKCC2 by stimulating exocytic insertion and that this pr

113 isms underlying the short term activation of NKCC2 by vasopressin in vivo, finding that administratio

114 brary through a yeast two-hybrid assay using NKCC2 C terminus as bait.

115 ney medullae, 2) a 150-amino acid stretch of NKCC2 C-terminal tail is involved in the interaction wit

116 tions in the apical Na-K-2Cl co-transporter, NKCC2, cause type I Bartter syndrome, a life-threatening

117 Here we report that NKCC2 co-immunoprecipitates with VAMP2 in rat TALs, and

118 cells, and stimulation of PKA enhanced VAMP2-NKCC2 co-immunoprecipitation in TALs.

119 SPAK activity and phosphorylation of NCC and NKCC2 co-transporters at the residues phosphorylated by

120 nd stimulating the Na-Cl (NCC) and Na-K-2Cl (NKCC2) co-transporters, which regulate salt reabsorption

121 s analysis revealed an amino acid stretch in NKCC2 containing apical sorting information.

122 We reported that constitutive endocytosis of NKCC2 controls NaCl absorption in native THALs; however,

123 among several analyzed motifs present in the NKCC2 COOH terminus, only those required for ER exit and

124 in the activation and surface expression of NKCC2 could play an important role in the regulated abso

125 imb (mTAL) apical Na+-K+-2Cl- cotransporter (NKCC2) decreased by 52 % (P < 0.02) and 44 % (P < 0.01),

126 VAMP2 was not involved in constitutive NKCC2 delivery.

127 Thus, NKCC2-dependent brain mechanisms that regulate osmotic s

128 a mechanism that may include alterations in NKCC2-dependent sodium reabsorption.

129 e kidney-specific NaK2Cl cotransporter, BSC1/NKCC2, do not survive.

130 Halving the mRNA expression of NKCC2 does not affect BP or fluid balance because of com

131 of a 4-gene signature of mRNAs for vimentin, NKCC2, E-cadherin, and 18S rRNA diagnostic of interstiti

132 Knockdown of HNS NKCC2 elicited profound effects on fluid balance followi

133 er: the absorptive isoform BSC1 (also called NKCC2, encoded by Slc12a1 in mouse) that is exclusively

134 dynamin-2, clathrin, and lipid rafts mediate NKCC2 endocytosis and maintain steady-state apical surfa

135 ive THALs; however, the pathways involved in NKCC2 endocytosis are unknown.

136 We hypothesized that NKCC2 endocytosis at the apical surface depends on dynam

137 egative Dyn2K44A in THALs slowed the rate of NKCC2 endocytosis by 38 +/- 8% and increased steady-stat

138 Disruption of lipid rafts blunted NKCC2 endocytosis by 39 +/- 4% and silencing caveolin-1

139 m interacting with synaptojanin also blunted NKCC2 endocytosis by 52 +/- 5%.

140 ated endocytosis with chlorpromazine blunted NKCC2 endocytosis by 54 +/- 6%, while preventing clathri

141 f endogenous dynamin-2 with dynasore blunted NKCC2 endocytosis by 56 +/- 11% and increased steady-sta

142 f steady-state surface NKCC2 and the rate of NKCC2 endocytosis in freshly isolated rat THALs showed t

143 identifying the endocytic pathway for apical NKCC2 endocytosis.

144 t VAMP3 selectively mediates cAMP-stimulated NKCC2 exocytic delivery and surface expression in TALs.

145 cAMP stimulates NKCC2 exocytic delivery via protein kinase A (PKA), incr

146 silencing VAMP3 in vivo blocks constitutive NKCC2 exocytic delivery, decreasing the amount of NKCC2

147 VAMP3 is not required for cAMP-stimulated NKCC2 exocytic delivery.

148 ssion and completely blocked cAMP-stimulated NKCC2 exocytic delivery.

149 ed cumulative apical membrane exocytosis and NKCC2 exocytic insertion in TALs.

150 The NKCC2 expressed in these mice was highly glycosylated an

151 se gene coding for the NaK2Cl cotransporter (NKCC2) expressed in kidney epithelial cells of the thick

152 Consequently, mice lacking AC6 have lower NKCC2 expression and a mild Bartter syndrome-like phenot

153 blunted cAMP-stimulated steady-state surface NKCC2 expression and completely blocked cAMP-stimulated

154 Renal AC6 determines total NKCC2 expression and mediates vasopressin-induced NKCC2/

155 wn of OS9 by small interfering RNA increased NKCC2 expression by increasing the co-transporter stabil

156 Additionally, VAMP3 is required for normal NKCC2 expression, renal function, and blood pressure.

157 e NKCC2 by 37 +/- 8%, without changing total NKCC2 expression.

158 ate p62 displayed the strongest reduction of NKCC2 expression.

159 teristics we expressed cDNAs encoding rabbit NKCC2 F, A, and B in Xenopus oocytes and determined the

160 variants of the renal Na-K-Cl cotransporter (NKCC2 F, A, and B) are spatially distributed along the t

161 oduces a premature stop in codon W625 of the NKCC2 gene (SCL12A1).

162 apical sorting of the protein encoded by the NKCC2 gene.

163 Serine-threonine kinases that activate NKCC2 have been identified, but less is known about phos

164 Confocal imaging of apical surface NKCC2 in isolated perfused TALs confirmed a stimulatory

165 stal nephron did not alter the expression of NKCC2 in mTAL and decreased AQP2 protein only in OM but

166 sis and maintain steady-state apical surface NKCC2 in native THALs.

167 pression of total and phosphorylated NCC and NKCC2 in respective nephron segments.

168 rapidly normalized the abundance of phospho-NKCC2 in SORLA-deficient mice, and a functional interact

169 lso found constitutive exocytic insertion of NKCC2 in TALs over time, which was increased by 3-fold i

170 NCC and the Na(+)-K(+)-2Cl(-) cotransporter NKCC2 in the distal convoluted tubule and the thick asce

171 Thus absence of NKCC2 in the mouse causes polyuria that is not compensat

172 on of vasopressin restored the expression of NKCC2 in the outer medulla as well as the expression and

173 VAMP3 in mice decreased total expression of NKCC2 in the TAL and lowered blood pressure.

174 There is no functional linking of ROMK1 and NKCC2 in the TAL.

175 n and no clear change in the distribution of NKCC2 in the thick ascending limb (TAL) cells.

176 he medullary Na(+)-K(+)-2Cl(-) cotransporter NKCC2 in these mice compared with wild-type mice, an eff

177 , KS-WNK1 is a negative regulator of NCC and NKCC2 in vivo and plays an important role in the control

178 glucose or NaCl, upregulated renal AQP2 and NKCC2 in vivo in BB rats.

179 plasmalemmal translocation and activation of NKCC2 in vivo.

180 glycosylation and cell surface expression of NKCC2, independently of the expression system.

181 mice that are deficient in the B isoform of NKCC2 indicates a limited role for NKCC2B for overall sa

182 Here we report that NKCC2 interacts with the vesicle fusion protein VAMP3, a

183 raft-mediated endocytosis completely blocked NKCC2 internalization.

184 Although a pool of NKCC2 is present in cytoplasmic vesicles, the distributi

185 Since NKCC2 is the molecular target of the loop diuretics bume

186 e-sensitive Na(+)-K(+)-2Cl(-)-cotransporter (NKCC2) is crucial for NaCl reabsorption in kidney thick

187 e-sensitive Na(+)-K(+)-2Cl(-) cotransporter (NKCC2) is responsible for urine concentration and helps

188 The renal Na-K-Cl cotransporter (NKCC2) is selectively expressed in the apical membranes

189 he renal-specific Na+-K+-2Cl- cotransporter (NKCC2) is the major salt transport pathway of the apical

190 AnxA2 effect involved only nonphosphorylated NKCC2, it appears to affect NKCC2 trafficking.

191 NKCC2 mRNA of NKCC2 +/- kidney was 55 +/- 6% of +/+, yet no difference

192 the renal specific Na-K-2Cl co-transporter (NKCC2) lead to type I Bartter syndrome, a life-threateni

193 Steady-state surface NKCC2 levels in the apical membrane are maintained by a

194 n of both high- and low-affinity isoforms of NKCC2 may permit transport and Cl-dependent tubuloglomer

195 Na-K-Cl cotransporter (NKCC2)-mediated sodium chloride reabsorption in the thic

196 ing of NKCC2 to the apical surface regulates NKCC2-mediated NaCl absorption and blood pressure.

197 se models suggest that OSR1 mainly activates NKCC2-mediated sodium transport along the thick ascendin

198 apical renal Na(+)-K(+)-2Cl(-) cotransporter NKCC2 mediates NaCl absorption by the thick ascending li

199 NKCC2 +/- mice had a near-normal level of NKCC2 protein

200 oscope images demonstrates a 55% increase in NKCC2 molecules at the apical membrane, suggesting the a

201 NKCC2 mRNA of NKCC2 +/- kidney was 55 +/- 6% of +/+, yet

202 The NKCC2 mutant animals should be valuable for uncovering n

203 NKCC2 mutations can be excluded in some Bartter's kindre

204 ntial phenotypes in heterozygous carriers of NKCC2 mutations.

205 In contrast, mutations of NKCC2 N-glycosylation sites abolished the effects of OS9

206 he abundance of total and cell-surface NHE3, NKCC2, NCC, alpha-ENaC and cleaved gamma-ENaC compared t

207 expression and mediates vasopressin-induced NKCC2/NCC phosphorylation.

208 transporters neither conferred activation to NKCC2 nor prevented activation of NKCC1.

209 that naturally occurring mutations depriving NKCC2 of its distal COOH-terminal tail and interfering w

210 strate that the differential distribution of NKCC2 on the proximal dendrites and KCC2 on the distal d

211 me residues with corresponding residues from NKCC2 or the Na-Cl cotransporter resulted in cation affi

212 bumetanide-sensitive Na-K-2Cl cotransporter (NKCC2) or the ATP-regulated potassium channel ROMK (KCNJ

213 ected with H(2)O or with RNA encoding SGLT1, NKCC2, or PepT1.

214 eptor with A-type repeats (SORLA) may affect NKCC2 phosphoregulation, we used SORLA-knockout mice to

215 a in the apical region of TAL cells and less NKCC2 phosphorylation and activity compared with litterm

216 increases and kinase-inactive WNK3 decreases NKCC2 phosphorylation at Thr-184 and Thr-189, sites requ

217 Thus, enhanced NKCC2 phosphorylation in the SPAK knock-out may be expla

218 in) causes a 2-fold increase in mouse kidney NKCC2 phosphorylation, as detected with a phosphospecifi

219 With regard to NKCC2 phosphorylation, the stimulatory effect of 1-desam

220 aporin 2 (AQP2) and Na-K-2Cl co-transporter (NKCC2), pivotal factors in urinary concentration, in AVP

221 ositive with NKCC1 primers and negative with NKCC2 primers.

222 AC6 is also a stimulator of total renal NKCC2 protein abundance in medullary and cortical thick

223 NKCC2 +/- mice had a near-normal level of NKCC2 protein and no clear change in the distribution of

224 In cells overexpressing OS9, total cellular NKCC2 protein levels were markedly decreased, an effect

225 by a reduction in the expression of NCC and NKCC2 protein without changes in mRNA levels.

226 athway in renal cells that degrades immature NKCC2 proteins.

227 Homozygous NKCC2-/- pups were born in expected numbers and appeared

228 The molecular mechanisms by which NKCC2 reaches the apical surface and their role in renal

229 levels of the apical Na/K/2Cl cotransporter NKCC2 regulate NaCl reabsorption by epithelial cells of

230 to investigate the effect of WNK3 on NCC and NKCC2, related kidney-specific transporters that mediate

231 NKCC2 rose in the ISOM but was not reversed with insulin

232 The three splice variants of NKCC2 showed dramatic differences in their kinetic behav

233 sodium-potassium -chloride co-transporter 2 (NKCC2), sodium chloride co-transporter (NCC), aquaporin

234 odium/potassium/chloride transporter type 2 (NKCC2), sodium/chloride transporter, and Na(+),K(+)-ATPa

235 Thus, the kinetic properties of the NKCC2 splice variants are consistent with the spatial di

236 To test whether NKCC2 splice variants differ in ion transport characteri

237 egulation of the expression of renal AQP and NKCC2, studies were performed with hyperosmolality that

238 e genes encoding the Na-K-2Cl cotransporter (NKCC2), the potassium channel ROMK, the chloride channel

239 Na,K,2Cl co-transporter (NKCC2), the primary NaCl uptake pathway in the thick asc

240 organize microdomains, is codistributed with NKCC2 to promote its apical translocation in response to

241 Trafficking of NKCC2 to the apical surface regulates NKCC2-mediated NaC

242 the mechanisms underlying the regulation of NKCC2 trafficking in renal cells are scarcely known.

243 onphosphorylated NKCC2, it appears to affect NKCC2 trafficking.

244 associated with a 60% decrease in medullary NKCC2 transporter expression determined by Western blot.

245 ammoniagenesis but reduced abundance of the NKCC2 transporter responsible for medullary accumulation

246 tation of the kinetic characteristics of the NKCC2 variants to the luminal concentrations of substrat

247 sence of the terminally glycosylated form of NKCC2 was not due to reduced synthesis or increased rate

248 The Na-K-2Cl cotransporter 2 (NKCC2) was thought to be kidney specific.

249 ing mechanism by which cAMP increases apical NKCC2, we measured cumulative apical membrane exocytosis

250 variants to interact with and phosphorylate NKCC2, whereas only full-length SPAK promoted the activa

251 (500 microm) stimulated steady-state surface NKCC2, whereas the Epac-selective agonist 8-p-chlorophen

252 een identified in the mammalian kidney: BSC1/NKCC2 which localizes to the apical thick ascending limb

253 tory epithelia and non-epithelial cells; and NKCC2, which is present exclusively in the kidney, in th

254 umulation in secretory epithelial cells, and NKCC2, which mediates apical Na+K+Cl entry into renal ep

255 se models suggest that OSR1 mainly activates NKCC2, while SPAK mainly activates NCC, with possible cr

256 Association of NKCC2 with lipid rafts facilitates its AVP-induced apica

257 mbrane domain 2 endows different versions of NKCC2 with unique kinetic behaviors.