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Author Notes:

To whom correspondence should be addressed: Dr. Robert S. Hoover, Division of Nephrology, Department of Medicine, Emory University, 615 Michael St, rm. 601, Atlanta, GA, 30322, Telephone: 404-727-2525; Fax: 404-727-3425. robert.hoover@emory.edu

Author Contributions: ACM and BMW conducted much of the experimentation, analyzed the results, interpreted the data and did some of the writing.

LY, VT, QY, YZ, OA, RM, AAA, AM, HFB, and FT also conducted experimentation and interpreted results.

BK, HC, OP, and DCE helped in interpretation of data and writing the paper.

DCE also helped conceive the paper.

RSH conceived the idea for the project and wrote most of the paper.

The authors would like to thank Jan Loffing for providing antibodies for the EM studies.

Declarations of Interest: The authors declare that they have no conflicts of interest with the content of this article.

Subjects:

Research Funding:

This work was supported by grants from the National Institutes of Health R01 DK-085097 (to R.S.H.), T32 DK07656 (B.M.W.), 1K01DK099617 (A.A.A.), R37 DK037963 (D.C.E.) and RO1 DK095029 (O.P.).

Also by Department of Veteran Affairs VA Merit Awards I01BX002322-01 (R.S.H.) and I01BX000994 (H.C.).

Also the Swiss National Foundation (F.T.), AHA GIA 13GRNT16220002 (O.P.), and the Russian Foundation for Basic Research #15-04-00938.

This research project was also supported in part by the Emory University Integrated Cellular Imaging Microscopy Core of the Emory University Department of Physiology.

Keywords:

  • Science & Technology
  • Life Sciences & Biomedicine
  • Biochemistry & Molecular Biology
  • NA-CL COTRANSPORTER
  • DISTAL CONVOLUTED TUBULE
  • TRANSPORT PATHWAYS
  • BLOOD-PRESSURE
  • MINERALOCORTICOID RECEPTORS
  • BARTTERS-SYNDROME
  • KNOCKOUT MICE
  • STAS DOMAIN
  • PROTEIN
  • EXPRESSION

The sodium chloride cotransporter (NCC) and epithelial sodium channel (ENaC) associate

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Journal Title:

Biochemical Journal

Volume:

Volume 473, Number 19

Publisher:

, Pages 3237-3252

Type of Work:

Article | Post-print: After Peer Review

Abstract:

The thiazide-sensitive sodium chloride cotransporter (NCC) and the epithelial sodium channel (ENaC) are two of the most important determinants of salt balance and thus systemic blood pressure. Abnormalities in either result in profound changes in blood pressure. There is one segment of the nephron where these two sodium transporters are coexpressed, the second part of the distal convoluted tubule. This is a key part of the aldosterone-sensitive distal nephron, the final regulator of salt handling in the kidney. Aldosterone is the key hormonal regulator for both of these proteins. Despite these shared regulators and coexpression in a key nephron segment, associations between these proteins have not been investigated. After confirming apical localization of these proteins, we demonstrated the presence of functional transport proteins and native association by blue native PAGE. Extensive coimmunoprecipitation experiments demonstrated a consistent interaction of NCC with α-And γ-ENaC. Mammalian two-hybrid studies demonstrated direct binding of NCC to ENaC subunits. Fluorescence resonance energy transfer and immunogold EM studies confirmed that these transport proteins are within appropriate proximity for direct binding. Additionally, we demonstrate that there are functional consequences of this interaction, with inhibition of NCC affecting the function of ENaC. This novel finding of an association between ENaC and NCC could alter our understanding of salt transport in the distal tubule.

Copyright information:

© 2016 The Author(s); published by Portland Press Limited on behalf of the Biochemical Society.

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