Regulation of maturation and processing of ENaC subunits in the rat kidney.
Antibodies directed against subunits of the epithelial Na channel (ENaC) were used together with electrophysiological measurements in the cortical collecting duct to investigate the processing of the proteins in rat kidney with changes in Na or K intake. When animals were maintained on a low-Na diet for 7-9 days, the abundance of two forms of the alpha-subunit, with apparent masses of 85 and 30 kDa, increased. Salt restriction also increased the abundance of the beta-subunit and produced an endoglycosidase H (Endo H)-resistant pool of this subunit. The abundance of the 90-kDa form of the gamma-subunit decreased, whereas that of a 70-kDa form increased and this peptide also exhibited Endo H-resistant glycosylation. These changes in alpha- and gamma-subunits were correlated with increases in Na conductance elicited by a 4-h infusion with aldosterone. Changes in all three subunits were correlated with decreases in Na conductance when Na-deprived animals drank saline for 5 h. We conclude that ENaC subunits are mainly in an immature form in salt-replete rats. With Na depletion, the subunits mature in a process that involves proteolytic cleavage and further glycosylation. Similar changes occurred in alpha- and gamma- but not beta-subunits when animals were treated with exogenous aldosterone, and in beta- and gamma- but not alpha-subunits when animals were fed a high-K diet. Changes in the processing and maturation of the channels occur rapidly enough to be involved in the daily regulation of ENaC activity and Na reabsorption by the kidney.
Department of Physiology and Biophysics, Weill Medical College of Cornell University, 1300 York Ave., New York, NY 10021, USA.
Zuhal
Z
Ergonul
Gustavo
G
Frindt
Lawrence G
LG
Palmer
D000450:Aldosterone / Q000494:pharmacology; D000818:Animals; D053503:Epithelial Sodium Channels; D005260:Female; D005786:Gene Expression Regulation / Q000502:physiology; D015151:Immunoblotting; D007668:Kidney / Q000187:drug effects / Q000378:metabolism*; D008297:Male; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase / Q000378:metabolism; D017419:Potassium, Dietary / Q000494:pharmacology; D021122:Protein Subunits / Q000235:genetics; D021381:Protein Transport / Q000502:physiology; D051381:Rats; D017207:Rats, Sprague-Dawley; D015222:Sodium Channels / Q000235:genetics*; D017673:Sodium Chloride, Dietary / Q000494:pharmacology
D053503:Epithelial Sodium Channels; D017419:Potassium, Dietary; D021122:Protein Subunits; D015222:Sodium Channels; D017673:Sodium Chloride, Dietary; D000450:Aldosterone; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase