Interaction of the Ca2+-sensing receptor with the inwardly rectifying potassium channels Kir4.1 and Kir4.2 results in inhibition of channel function

Chunfa Huang, Aleksandra Sindic, Ceredwyn E. Hill, Kristine M. Hujer, Kim W. Chan, Martin Sassen, Zhenzhen Wu, Yoshihisa Kurachi, Soren Nielsen, Michael F. Romero, R. Tyler Miller

Research output: Contribution to journalArticle

60 Citations (Scopus)

Abstract

The Ca2+-sensing receptor (CaR), a G protein-coupled receptor, is expressed in many epithelial tissues including the parathyroid glands, kidney, and GI tract. Although its role in regulating PTH levels and Ca 2+ metabolism are best characterized, it may also regulate salt and water transport in the kidney as demonstrated by recent reports showing association of potent gain-of-function mutations in the CaR with a Bartter-like, salt-wasting phenotype. To determine whether this receptor interacts with novel proteins that control ion transport, we screened a human adult kidney cDNA library with the COOH-terminal 219 amino acid cytoplasmic tail of the CaR as bait using the yeast two-hybrid system. We identified two independent clones coding for ∼125 aa from the COOH terminus of the inwardly rectifying K + channel, Kir4.2. The CaR and Kir4.2 as well as Kir4.1 (another member of Kir4 subfamily) were reciprocally coimmunoprecipitated from HEK-293 cells in which they were expressed, but the receptor did not coimmunoprecipitate with Kir5.1 or Kir1.1. Both Kir4.1 and Kir4.2 were immunoprecipitated from rat kidney extracts with the CaR. In Xenopus laevis oocytes, expression of the CaR with either Kir4.1 or Kir4.2 channels resulted in inactivation of whole cell current as measured by two-electrode voltage clamp, but the nonfunctional CaR mutant CaRR796W, and that does not coimmunoprecipitate with the channels, had no effect. Kir4.1 and the CaR were colocalized in the basolateral membrane of the distal nephron. The CaR interacts directly with Kir4.1 and Kir4.2 and can decrease their currents, which in turn could reduce recycling of K+ for the basolateral Na+-K+-ATPase and thereby contribute to inhibition of Na+ reabsorption.

Original languageEnglish (US)
JournalAmerican Journal of Physiology - Renal Physiology
Volume292
Issue number3
DOIs
StatePublished - Mar 2007

Fingerprint

Inwardly Rectifying Potassium Channel
Calcium-Sensing Receptors
Kidney
Salts
Two-Hybrid System Techniques
Parathyroid Glands
HEK293 Cells
Nephrons
Ion Transport
Xenopus laevis
Recycling
G-Protein-Coupled Receptors
Gene Library
Oocytes
Gastrointestinal Tract
Tail
Electrodes
Epithelium
Clone Cells
Phenotype

Keywords

  • HEK 293 cells
  • Inwardly rectifying K channel
  • Yeast two hybrid

ASJC Scopus subject areas

  • Physiology

Cite this

Interaction of the Ca2+-sensing receptor with the inwardly rectifying potassium channels Kir4.1 and Kir4.2 results in inhibition of channel function. / Huang, Chunfa; Sindic, Aleksandra; Hill, Ceredwyn E.; Hujer, Kristine M.; Chan, Kim W.; Sassen, Martin; Wu, Zhenzhen; Kurachi, Yoshihisa; Nielsen, Soren; Romero, Michael F.; Miller, R. Tyler.

In: American Journal of Physiology - Renal Physiology, Vol. 292, No. 3, 03.2007.

Research output: Contribution to journalArticle

Huang, Chunfa ; Sindic, Aleksandra ; Hill, Ceredwyn E. ; Hujer, Kristine M. ; Chan, Kim W. ; Sassen, Martin ; Wu, Zhenzhen ; Kurachi, Yoshihisa ; Nielsen, Soren ; Romero, Michael F. ; Miller, R. Tyler. / Interaction of the Ca2+-sensing receptor with the inwardly rectifying potassium channels Kir4.1 and Kir4.2 results in inhibition of channel function. In: American Journal of Physiology - Renal Physiology. 2007 ; Vol. 292, No. 3.
@article{6c2d6aea8c7f4586a7131f03e81eb4b2,
title = "Interaction of the Ca2+-sensing receptor with the inwardly rectifying potassium channels Kir4.1 and Kir4.2 results in inhibition of channel function",
abstract = "The Ca2+-sensing receptor (CaR), a G protein-coupled receptor, is expressed in many epithelial tissues including the parathyroid glands, kidney, and GI tract. Although its role in regulating PTH levels and Ca 2+ metabolism are best characterized, it may also regulate salt and water transport in the kidney as demonstrated by recent reports showing association of potent gain-of-function mutations in the CaR with a Bartter-like, salt-wasting phenotype. To determine whether this receptor interacts with novel proteins that control ion transport, we screened a human adult kidney cDNA library with the COOH-terminal 219 amino acid cytoplasmic tail of the CaR as bait using the yeast two-hybrid system. We identified two independent clones coding for ∼125 aa from the COOH terminus of the inwardly rectifying K + channel, Kir4.2. The CaR and Kir4.2 as well as Kir4.1 (another member of Kir4 subfamily) were reciprocally coimmunoprecipitated from HEK-293 cells in which they were expressed, but the receptor did not coimmunoprecipitate with Kir5.1 or Kir1.1. Both Kir4.1 and Kir4.2 were immunoprecipitated from rat kidney extracts with the CaR. In Xenopus laevis oocytes, expression of the CaR with either Kir4.1 or Kir4.2 channels resulted in inactivation of whole cell current as measured by two-electrode voltage clamp, but the nonfunctional CaR mutant CaRR796W, and that does not coimmunoprecipitate with the channels, had no effect. Kir4.1 and the CaR were colocalized in the basolateral membrane of the distal nephron. The CaR interacts directly with Kir4.1 and Kir4.2 and can decrease their currents, which in turn could reduce recycling of K+ for the basolateral Na+-K+-ATPase and thereby contribute to inhibition of Na+ reabsorption.",
keywords = "HEK 293 cells, Inwardly rectifying K channel, Yeast two hybrid",
author = "Chunfa Huang and Aleksandra Sindic and Hill, {Ceredwyn E.} and Hujer, {Kristine M.} and Chan, {Kim W.} and Martin Sassen and Zhenzhen Wu and Yoshihisa Kurachi and Soren Nielsen and Romero, {Michael F.} and Miller, {R. Tyler}",
year = "2007",
month = "3",
doi = "10.1152/ajprenal.00269.2006",
language = "English (US)",
volume = "292",
journal = "American Journal of Physiology - Heart and Circulatory Physiology",
issn = "0363-6135",
publisher = "American Physiological Society",
number = "3",

}

TY - JOUR

T1 - Interaction of the Ca2+-sensing receptor with the inwardly rectifying potassium channels Kir4.1 and Kir4.2 results in inhibition of channel function

AU - Huang, Chunfa

AU - Sindic, Aleksandra

AU - Hill, Ceredwyn E.

AU - Hujer, Kristine M.

AU - Chan, Kim W.

AU - Sassen, Martin

AU - Wu, Zhenzhen

AU - Kurachi, Yoshihisa

AU - Nielsen, Soren

AU - Romero, Michael F.

AU - Miller, R. Tyler

PY - 2007/3

Y1 - 2007/3

N2 - The Ca2+-sensing receptor (CaR), a G protein-coupled receptor, is expressed in many epithelial tissues including the parathyroid glands, kidney, and GI tract. Although its role in regulating PTH levels and Ca 2+ metabolism are best characterized, it may also regulate salt and water transport in the kidney as demonstrated by recent reports showing association of potent gain-of-function mutations in the CaR with a Bartter-like, salt-wasting phenotype. To determine whether this receptor interacts with novel proteins that control ion transport, we screened a human adult kidney cDNA library with the COOH-terminal 219 amino acid cytoplasmic tail of the CaR as bait using the yeast two-hybrid system. We identified two independent clones coding for ∼125 aa from the COOH terminus of the inwardly rectifying K + channel, Kir4.2. The CaR and Kir4.2 as well as Kir4.1 (another member of Kir4 subfamily) were reciprocally coimmunoprecipitated from HEK-293 cells in which they were expressed, but the receptor did not coimmunoprecipitate with Kir5.1 or Kir1.1. Both Kir4.1 and Kir4.2 were immunoprecipitated from rat kidney extracts with the CaR. In Xenopus laevis oocytes, expression of the CaR with either Kir4.1 or Kir4.2 channels resulted in inactivation of whole cell current as measured by two-electrode voltage clamp, but the nonfunctional CaR mutant CaRR796W, and that does not coimmunoprecipitate with the channels, had no effect. Kir4.1 and the CaR were colocalized in the basolateral membrane of the distal nephron. The CaR interacts directly with Kir4.1 and Kir4.2 and can decrease their currents, which in turn could reduce recycling of K+ for the basolateral Na+-K+-ATPase and thereby contribute to inhibition of Na+ reabsorption.

AB - The Ca2+-sensing receptor (CaR), a G protein-coupled receptor, is expressed in many epithelial tissues including the parathyroid glands, kidney, and GI tract. Although its role in regulating PTH levels and Ca 2+ metabolism are best characterized, it may also regulate salt and water transport in the kidney as demonstrated by recent reports showing association of potent gain-of-function mutations in the CaR with a Bartter-like, salt-wasting phenotype. To determine whether this receptor interacts with novel proteins that control ion transport, we screened a human adult kidney cDNA library with the COOH-terminal 219 amino acid cytoplasmic tail of the CaR as bait using the yeast two-hybrid system. We identified two independent clones coding for ∼125 aa from the COOH terminus of the inwardly rectifying K + channel, Kir4.2. The CaR and Kir4.2 as well as Kir4.1 (another member of Kir4 subfamily) were reciprocally coimmunoprecipitated from HEK-293 cells in which they were expressed, but the receptor did not coimmunoprecipitate with Kir5.1 or Kir1.1. Both Kir4.1 and Kir4.2 were immunoprecipitated from rat kidney extracts with the CaR. In Xenopus laevis oocytes, expression of the CaR with either Kir4.1 or Kir4.2 channels resulted in inactivation of whole cell current as measured by two-electrode voltage clamp, but the nonfunctional CaR mutant CaRR796W, and that does not coimmunoprecipitate with the channels, had no effect. Kir4.1 and the CaR were colocalized in the basolateral membrane of the distal nephron. The CaR interacts directly with Kir4.1 and Kir4.2 and can decrease their currents, which in turn could reduce recycling of K+ for the basolateral Na+-K+-ATPase and thereby contribute to inhibition of Na+ reabsorption.

KW - HEK 293 cells

KW - Inwardly rectifying K channel

KW - Yeast two hybrid

UR - http://www.scopus.com/inward/record.url?scp=33847771725&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33847771725&partnerID=8YFLogxK

U2 - 10.1152/ajprenal.00269.2006

DO - 10.1152/ajprenal.00269.2006

M3 - Article

C2 - 17122384

AN - SCOPUS:33847771725

VL - 292

JO - American Journal of Physiology - Heart and Circulatory Physiology

JF - American Journal of Physiology - Heart and Circulatory Physiology

SN - 0363-6135

IS - 3

ER -