A single residue in transmembrane domain 11 defines the different affinity for thiazides between the mammalian and flounder NaCl transporters

Castañeda‐Bueno, María; Vázquez, Norma; Bustos‐Jaimes, Ismael; Hernandez, Damian; Rodríguez-Lobato, Erika; Pacheco‐Alvarez, Diana; Cariño-Cortés, Raquel; Moreno, Erika; Bobadilla, Norma A.; Gamba, Gerardo

doi:10.1152/ajprenal.00412.2010

Cited by 22 publications

(16 citation statements)

References 24 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Site-directed mutagenesis was used to show that a single residue in TM11 defines the different affinity for thiazides between mammalian and flounder NCC [12]. Both transmembrane regions appear to have affinity for extracellular sodium.…”

Section: Structure-function Relationshipmentioning

confidence: 99%

The sodium chloride cotransporter SLC12A3: new roles in sodium, potassium, and blood pressure regulation

Moes

Lubbe

Zietse

et al. 2013

Pflugers Arch - Eur J Physiol

View full text Add to dashboard Cite

SLC12A3 encodes the thiazide-sensitive sodium chloride cotransporter (NCC), which is primarily expressed in the kidney, but also in intestine and bone. In the kidney, NCC is located in the apical plasma membrane of epithelial cells in the distal convoluted tubule. Although NCC reabsorbs only 5 to 10 % of filtered sodium, it is important for the fine-tuning of renal sodium excretion in response to various hormonal and non-hormonal stimuli. Several new roles for NCC in the regulation of sodium, potassium, and blood pressure have been unraveled recently. For example, the recent discoveries that NCC is activated by angiotensin II but inhibited by dietary potassium shed light on how the kidney handles sodium during hypovolemia (high angiotensin II) and hyperkalemia. The additive effect of angiotensin II and aldosterone maximizes sodium reabsorption during hypovolemia, whereas the inhibitory effect of potassium on NCC increases delivery of sodium to the potassium-secreting portion of the nephron. In addition, great steps have been made in unraveling the molecular machinery that controls NCC. This complex network consists of kinases and ubiquitinases, including WNKs, SGK1, SPAK, Nedd4-2, Cullin-3, and Kelch-like 3. The pathophysiological significance of this network is illustrated by the fact that modification of each individual protein in the network changes NCC activity and results in salt-dependent hypotension or hypertension. This review aims to summarize these new insights in an integrated manner while identifying unanswered questions.

show abstract

Section: Structure-function Relationshipmentioning

confidence: 99%

The sodium chloride cotransporter SLC12A3: new roles in sodium, potassium, and blood pressure regulation

Moes

Lubbe

Zietse

et al. 2013

Pflugers Arch - Eur J Physiol

View full text Add to dashboard Cite

show abstract

“…Prior to the uptake assay, oocytes were switched to Cl --free ND96 for 2-16 hours or to low-chloride hypotonic stress medium for 16 hours, as previously described [24,28]. [11,24,27,29]. Activation of NCC by low chloride hypotonic stress was done as previously described [28].…”

Section: Preparation Of Xenopus Laevis Oocytesmentioning

confidence: 99%

WNK3-SPAK Interaction is Required for the Modulation of NCC and other Members of the SLC12 Family

Pacheco‐Alvarez

Vázquez

Castañeda‐Bueno

et al. 2012

Cell Physiol Biochem

Self Cite

View full text Add to dashboard Cite

The serine/threonine with no lysine kinase 3 (WNK3) modulates the activity of the electroneutral cation-coupled chloride cotransporters (CCC) to promote Cl^- influx and prevent Cl^- efflux, thus fitting the profile for a putative “Cl^--sensing kinase”. The Ste20-type kinases, SPAK/OSR1, become phosphorylated in response to reduction in intracellular chloride concentration and regulate the activity of NKCC1. Several studies have now shown that WNKs function upstream of SPAK/OSR1. This study was designed to analyze the role of WNK3-SPAK interaction in the regulation of CCCs with particular emphasis on NCC. In this study we used the functional expression system of Xenopus laevis oocytes to show that different SPAK binding sites in WNK3 (^{241, 872, 1336}RFxV) are required for the kinase to have effects on CCCs. WNK3-F1337A no longer activated NKCC2, but the effects on NCC, NKCC1, and KCC4 were preserved. In contrast, the effects of WNK3 on these cotransporters were prevented in WNK3-F242A. The elimination of F873 had no consequence on WNK3 effects. WNK3 promoted NCC phosphorylation at threonine 58, even in the absence of the unique SPAK binding site of NCC, but this effect was abolished in the mutant WNK3-F242A. Thus, our data support the hypothesis that the effects of WNK3 upon NCC and other CCCs require the interaction and activation of the SPAK kinase. The effect is dependent on one of the three binding sites for SPAK that are present in WNK3, but not on the SPAK binding sites on the CCCs, which suggests that WNK3 is capable of binding both SPAK and CCCs to promote their phosphorylation.

show abstract

“…After 2-3 days of incubation, metolazone-sensitive 22 Na ϩ (for NCC), bumetanide-sensitive 86 Rb ϩ (for NKCC1 and NKCC2), or Cl Ϫ -dependent 86 Rb ϩ uptake (for KCCs) was determined following our standard procedures (1,2,17,20). In brief, NCC measurements were performed under isotonic conditions.…”

Section: Functional Assays With Cation/cl ϫ Cotransportersmentioning

confidence: 99%

Similar effects of all WNK3 variants on SLC12 cotransporters

Cruz-Rangel

Melo

Vázquez

et al. 2011

American Journal of Physiology-Cell Physiology

Self Cite

View full text Add to dashboard Cite

kinase 3 (WNK3) is a member of a subfamily of serine/threonine kinases that modulate the activity of the electroneutral cation-coupled chloride cotransporters. WNK3 activates NKCC1/2 and NCC and inhibits the KCCs. Four splice variants are generated from the WNK3 gene. Our previous studies focused on the WNK3-18a variant. However, it has been suggested that other variants could have different effects on the cotransporters. Thus, the present study was designed to define the effects of all WNK3 variants on members of the SLC12 family. By RT-PCR from a fetal brain library, exons 18b and 22 were separately amplified and subcloned into the original WNK3-18a or catalytically inactive WNK3-D294A to obtain all four potential combinations with and without catalytic activity (18a, 18aϩ22, 18b, and 18bϩ22). The basal activity of the cotransporters and the effects of WNK3 isoforms were assessed in Xenopus laevis oocytes coinjected with each of the WNK3 variant cRNAs. In isotonic conditions, the basal activity of NCC and NKCC1/2 were increased by coinjection with any of the WNK3. The positive effects occurred even in hypotonic conditions, in which the basal activity of NKCC1 is completely prevented. Consistent with these observations, when expressed in hypotonicity, all KCCs were active, but in the presence of any of the WNK3 variants, KCC activity was completely reduced. That is, NKCC1/2 and NCC were inhibited, even in hypertonicity, while KCCs were activated, even in isotonic conditions. We conclude that the effects of all WNK3 variants toward SLC12 proteins are similar.

show abstract

A single residue in transmembrane domain 11 defines the different affinity for thiazides between the mammalian and flounder NaCl transporters

Cited by 22 publications

References 24 publications

The sodium chloride cotransporter SLC12A3: new roles in sodium, potassium, and blood pressure regulation

The sodium chloride cotransporter SLC12A3: new roles in sodium, potassium, and blood pressure regulation

WNK3-SPAK Interaction is Required for the Modulation of NCC and other Members of the SLC12 Family

Similar effects of all WNK3 variants on SLC12 cotransporters

Contact Info

Product

Resources

About