Interaction with grp58 increases activity of the thiazide-sensitive Na-Cl cotransporter

Bruce Wyse, Nawb Ali, David Ellison

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

The thiazide-sensitive sodium-chloride cotransporter (NCC) is expressed by distal convoluted tubule cells of the mammalian kidney. We used yeast two-hybrid screening to identify that glucose-regulated protein 58 (grp58), a protein induced by glucose deprivation, binds to the COOH terminus of the NCC. Immunoprecipitation of rat kidney cortex homogenates using a guinea pig anti-NCC antibody confirmed that grp58 associates with the NCC in vivo. Northern blots indicated that grp58 is highly expressed in rat kidney cortex. Immunofluorescence showed that grp58 protein abundance in kidney is highest in epithelial cells of the distal nephron, where it colocalizes with NCC near the apical membrane. To determine whether this interaction has a functional significance, NCC and grp58 cRNA were coexpressed in Xenopus laevis oocytes. In oocytes overexpressing grp58, sodium uptake was increased compared with control. Because oocytes express endogenous grp58, antisense experiments were performed to evaluate whether endogenous grp58 affected NCC activity in oocytes. Sodium uptake was lower in oocytes injected with both antisense grp58 cRNA and sense NCC compared with sense NCC oocytes. Western blot analysis did not show any effect of grp58 expression on processing of the NCC. These data indicate a novel, functionally important interaction between grp58 and the NCC in rat kidney cortex.

Original languageEnglish (US)
JournalAmerican Journal of Physiology - Renal Physiology
Volume282
Issue number3 51-3
StatePublished - 2002

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Oocytes
Kidney Cortex
Complementary RNA
thiazide receptor
glucose-regulated proteins
Member 3 Solute Carrier Family 12
Sodium
Kidney
Nephrons
Xenopus laevis
Immunoprecipitation
Northern Blotting
Fluorescent Antibody Technique
Anti-Idiotypic Antibodies
Guinea Pigs
Proteins
Yeasts
Western Blotting
Epithelial Cells
Glucose

Keywords

  • Distal
  • Glucose-regulated protein 58
  • Kidney tubules
  • Molecular chaperones
  • Thiazide-sensitive sodium-chloride co-transporter
  • Two-hybrid system techniques

ASJC Scopus subject areas

  • Physiology

Cite this

Interaction with grp58 increases activity of the thiazide-sensitive Na-Cl cotransporter. / Wyse, Bruce; Ali, Nawb; Ellison, David.

In: American Journal of Physiology - Renal Physiology, Vol. 282, No. 3 51-3, 2002.

Research output: Contribution to journalArticle

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