Chloride sensing by WNK1 involves inhibition of autophosphorylation

Alexander T. Piala, Thomas M. Moon, Radha Akella, Haixia He, Melanie H. Cobb, Elizabeth J. Goldsmith

Research output: Contribution to journalArticle

128 Citations (Scopus)

Abstract

WNK1 [with no lysine (K)] is a serine-threonine kinase associated with a form of familial hypertension. WNK1 is at the top of a kinase cascade, leading to phosphorylation of several cotransporters, in particular those transporting sodium, potassium, and chloride (NKCC), sodium and chloride (NCC), and potassium and chloride (KCC). The responsiveness of NKCC, NCC, and KCC to changes in extracellular chloride parallels their phosphorylation state, provoking the proposal that these transporters are controlled by a chloride-sensitive protein kinase. We found that chloride stabilizes the inactive conformation of WNK1, preventing kinase autophosphorylation and activation. Crystallographic studies of inactive WNK1 in the presence of chloride revealed that chloride binds directly to the catalytic site, providing a basis for the unique position of the catalytic lysine. Mutagenesis of the chloride-binding site rendered the kinase less sensitive to inhibition of autophosphorylation by chloride, validating the binding site. Thus, these data suggest that WNK1 functions as a chloride sensor through direct binding of a regulatory chloride ion to the active site, which inhibits autophosphorylation.

Original languageEnglish (US)
Article numberra41
JournalScience Signaling
Volume7
Issue number324
DOIs
StatePublished - May 6 2014

Fingerprint

Chlorides
Sodium Chloride
Phosphorylation
Phosphotransferases
Potassium Chloride
Lysine
Catalytic Domain
Binding Sites
Mutagenesis
Protein-Serine-Threonine Kinases
Protein Kinases
Conformations
Sodium
Chemical activation
Ions
Hypertension
Sensors

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology
  • Medicine(all)

Cite this

Chloride sensing by WNK1 involves inhibition of autophosphorylation. / Piala, Alexander T.; Moon, Thomas M.; Akella, Radha; He, Haixia; Cobb, Melanie H.; Goldsmith, Elizabeth J.

In: Science Signaling, Vol. 7, No. 324, ra41, 06.05.2014.

Research output: Contribution to journalArticle

@article{4e99d9c531a446299439e58597db2e39,
title = "Chloride sensing by WNK1 involves inhibition of autophosphorylation",
abstract = "WNK1 [with no lysine (K)] is a serine-threonine kinase associated with a form of familial hypertension. WNK1 is at the top of a kinase cascade, leading to phosphorylation of several cotransporters, in particular those transporting sodium, potassium, and chloride (NKCC), sodium and chloride (NCC), and potassium and chloride (KCC). The responsiveness of NKCC, NCC, and KCC to changes in extracellular chloride parallels their phosphorylation state, provoking the proposal that these transporters are controlled by a chloride-sensitive protein kinase. We found that chloride stabilizes the inactive conformation of WNK1, preventing kinase autophosphorylation and activation. Crystallographic studies of inactive WNK1 in the presence of chloride revealed that chloride binds directly to the catalytic site, providing a basis for the unique position of the catalytic lysine. Mutagenesis of the chloride-binding site rendered the kinase less sensitive to inhibition of autophosphorylation by chloride, validating the binding site. Thus, these data suggest that WNK1 functions as a chloride sensor through direct binding of a regulatory chloride ion to the active site, which inhibits autophosphorylation.",
author = "Piala, {Alexander T.} and Moon, {Thomas M.} and Radha Akella and Haixia He and Cobb, {Melanie H.} and Goldsmith, {Elizabeth J.}",
year = "2014",
month = "5",
day = "6",
doi = "10.1126/scisignal.2005050",
language = "English (US)",
volume = "7",
journal = "Science Signaling",
issn = "1937-9145",
publisher = "American Association for the Advancement of Science",
number = "324",

}

TY - JOUR

T1 - Chloride sensing by WNK1 involves inhibition of autophosphorylation

AU - Piala, Alexander T.

AU - Moon, Thomas M.

AU - Akella, Radha

AU - He, Haixia

AU - Cobb, Melanie H.

AU - Goldsmith, Elizabeth J.

PY - 2014/5/6

Y1 - 2014/5/6

N2 - WNK1 [with no lysine (K)] is a serine-threonine kinase associated with a form of familial hypertension. WNK1 is at the top of a kinase cascade, leading to phosphorylation of several cotransporters, in particular those transporting sodium, potassium, and chloride (NKCC), sodium and chloride (NCC), and potassium and chloride (KCC). The responsiveness of NKCC, NCC, and KCC to changes in extracellular chloride parallels their phosphorylation state, provoking the proposal that these transporters are controlled by a chloride-sensitive protein kinase. We found that chloride stabilizes the inactive conformation of WNK1, preventing kinase autophosphorylation and activation. Crystallographic studies of inactive WNK1 in the presence of chloride revealed that chloride binds directly to the catalytic site, providing a basis for the unique position of the catalytic lysine. Mutagenesis of the chloride-binding site rendered the kinase less sensitive to inhibition of autophosphorylation by chloride, validating the binding site. Thus, these data suggest that WNK1 functions as a chloride sensor through direct binding of a regulatory chloride ion to the active site, which inhibits autophosphorylation.

AB - WNK1 [with no lysine (K)] is a serine-threonine kinase associated with a form of familial hypertension. WNK1 is at the top of a kinase cascade, leading to phosphorylation of several cotransporters, in particular those transporting sodium, potassium, and chloride (NKCC), sodium and chloride (NCC), and potassium and chloride (KCC). The responsiveness of NKCC, NCC, and KCC to changes in extracellular chloride parallels their phosphorylation state, provoking the proposal that these transporters are controlled by a chloride-sensitive protein kinase. We found that chloride stabilizes the inactive conformation of WNK1, preventing kinase autophosphorylation and activation. Crystallographic studies of inactive WNK1 in the presence of chloride revealed that chloride binds directly to the catalytic site, providing a basis for the unique position of the catalytic lysine. Mutagenesis of the chloride-binding site rendered the kinase less sensitive to inhibition of autophosphorylation by chloride, validating the binding site. Thus, these data suggest that WNK1 functions as a chloride sensor through direct binding of a regulatory chloride ion to the active site, which inhibits autophosphorylation.

UR - http://www.scopus.com/inward/record.url?scp=84900543727&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84900543727&partnerID=8YFLogxK

U2 - 10.1126/scisignal.2005050

DO - 10.1126/scisignal.2005050

M3 - Article

VL - 7

JO - Science Signaling

JF - Science Signaling

SN - 1937-9145

IS - 324

M1 - ra41

ER -