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Intracellular Sodium Regulates Proteolytic Activation of the Epithelial Sodium Channel
Journal article   Open access   Peer reviewed

Intracellular Sodium Regulates Proteolytic Activation of the Epithelial Sodium Channel

Kristin K Knight, Danielle M Wentzlaff and Peter M Snyder
The Journal of biological chemistry, Vol.283(41), pp.27477-27482
10/10/2008
DOI: 10.1074/jbc.M804176200
PMCID: PMC2562069
PMID: 18662987
url
https://doi.org/10.1074/jbc.M804176200View
Published (Version of record) Open Access

Abstract

Na + transport across epithelia is mediated in part by the epithelial Na + channel ENaC. Previous work indicates that Na + is an important regulator of ENaC, providing a negative feedback mechanism to maintain Na + homeostasis. ENaC is synthesized as an inactive precursor, which is activated by proteolytic cleavage of the extracellular domains of the α and γ subunits. Here we found that Na + regulates ENaC in part by altering proteolytic activation of the channel. When the Na + concentration was low, we found that the majority of ENaC at the cell surface was in the cleaved/active state. As Na + increased, there was a dose-dependent decrease in ENaC cleavage and, hence, ENaC activity. This Na + effect was dependent on Na + permeation; cleavage was increased by the ENaC blocker amiloride and by a mutation that decreases ENaC activity (α H69A ) and was reduced by a mutation that activates ENaC (β S520K ). Moreover, the Na + ionophore monensin reversed the effect of the inactivating mutation (α H69A ) on ENaC cleavage, suggesting that intracellular Na + regulates cleavage. Na + did not alter activity of Nedd4-2, an E3 ubiquitin ligase that modulates ENaC cleavage, but Na + reduced ENaC cleavage by exogenous trypsin. Our findings support a model in which intracellular Na + regulates cleavage by altering accessibility of ENaC cleavage sites to proteases and provide a molecular explanation for the earlier observation that intracellular Na + inhibits Na + transport via ENaC (Na + feedback inhibition).
Membrane Transport, Structure, Function, and Biogenesis

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