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Author Notes:

E-mail Address : inyeong.choi@emory.edu

We acknowledge Eunjung Shin for construction of GST/syntrophin γ2 fusion proteins, Alana Darcher and Brigid Choi for art work and editorial correction.

We also thank Dr. Vincenzo Nigro (Seconda Università degli Studi di Napoli, Italy) for the syntrophin γ2 antibody.

Conflicts of Interests: None declared.

Subjects:

Research Funding:

This study was supported by National Institutes of Health grants NS055179 (Randy Hall), DK061418 (Chris Yun), and National Institutes of Health GM078502 and American Heart Association (Inyeong Choi).

Keywords:

  • Bicarbonate transporter
  • pH
  • protein interaction
  • SLC4A7
  • Xenopus oocyte

Systematic family-wide analysis of sodium bicarbonate cotransporter NBCn1/SLC4A7 interactions with PDZ scaffold proteins

Tools:

Journal Title:

Physiological Reports

Volume:

Volume 2, Number 5

Publisher:

, Pages e12016-e12016

Type of Work:

Article | Final Publisher PDF

Abstract:

NBCn1 (SLC4A7) plays a role in transepithelial HCO3− movement and intracellular pH maintenance in many tissues. In this study, we searched PDZ proteins capable of binding to NBCn1. We screened a protein array membrane, on which 96 different class I PDZ protein peptides were blotted, with the C‐terminal domain of NBCn1 fused to GST. Thirteen proteins were identified in these screens: MAGI‐3, NHERF‐1, NHERF‐2, PSD‐95, chapsyn‐110, ERBIN, MALS‐1, densin‐180, syntrophins α1, β2, γ2, MUPP1, and PDZK1. After determining these binding partners, we analyzed the database of known and predicted protein interactions to obtain an NBCn1 interaction network. The network shows NBCn1 being physically and functionally associated with a variety of membrane and cytosolic proteins via the binding partners. We then focused on syntrophin γ2 to examine the molecular and functional interaction between NBCn1 and one of the identified binding partners in the Xenopus oocyte expression system. GST/NBCn1 pulled down syntrophin γ2 and conversely GST/syntrophin γ2 pulled down NBCn1. Moreover, syntrophin γ2 increased intracellular pH recovery, from acidification, mediated by NBCn1's Na/HCO3 cotransport. Syntrophin γ2 also increased an ionic conductance produced by NBCn1 channel‐like activity. Thus, syntrophin γ2 regulates NBCn1 activity. In conclusion, this study demonstrates that NBCn1 binds to many PDZ proteins, which in turn may allow the transporter to associate with other physiologically important proteins.

Copyright information:

© 2014 The Authors. Physiological Reports published by Wiley Periodicals, Inc. on behalf of the American Physiological Society and The Physiological Society.

This is an Open Access work distributed under the terms of the Creative Commons Attribution 3.0 Unported License (http://creativecommons.org/licenses/by/3.0/).

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