title: Carboxylmethylation of the beta subunit of xENaC regulates channel activity.
abstract: The action of aldosterone to increase apical membrane permeability in responsive epithelia is thought to be due to activation of sodium channels. Aldosterone stimulates methylation of a 95-kDa protein in apical membrane of A6 cells, and we have previously shown that methylation of a 95-kDa protein in the immunopurified Na+ channel complex increases open probability of these channels in planar lipid bilayers. We report here that aldosterone stimulates carboxylmethylation of the beta subunit of xENaC in A6 cells. In vitro translated beta subunit, but not alpha or gamma, serves as a substrate for carboxylmethylation. Carboxylmethylation of ENaC reconstituted in planar lipid bilayers leads to an increase in open probability only when beta subunit is present. When the channel complex is immunoprecipitated from A6 cells and analyzed by Western blot with antibodies to the three subunits of xENaC, all three subunits are recognized as constituents of the complex. The results suggest that Na+ channel activity in A6 cells is regulated, in part, by carboxylmethylation of the beta subunit of xENaC.
authors.affiliation: Laboratory of Epithelial Cell Biology, Department of Medicine, University of Pittsburgh, Pittsburgh, Pennsylvania 15213, USA.
authors.forename: M D
authors.initials: MD
authors.lastname: Rokaw
authors.forename: J M
authors.initials: JM
authors.lastname: Wang
authors.forename: R S
authors.initials: RS
authors.lastname: Edinger
authors.forename: O A
authors.initials: OA
authors.lastname: Weisz
authors.forename: D
authors.initials: D
authors.lastname: Hui
authors.forename: P
authors.initials: P
authors.lastname: Middleton
authors.forename: V
authors.initials: V
authors.lastname: Shlyonsky
authors.forename: B K
authors.initials: BK
authors.lastname: Berdiev
authors.forename: I
authors.initials: I
authors.lastname: Ismailov
authors.forename: D C
authors.initials: DC
authors.lastname: Eaton
authors.forename: D J
authors.initials: DJ
authors.lastname: Benos
authors.forename: J P
authors.initials: JP
authors.lastname: Johnson
mesh_terms: D000450:Aldosterone / Q000494:pharmacology; D000595:Amino Acid Sequence; D000906:Antibodies / Q000276:immunology; D002460:Cell Line; D004591:Electrophoresis, Polyacrylamide Gel; D008051:Lipid Bilayers; D008745:Methylation; D015222:Sodium Channels / Q000276:immunology / Q000378:metabolism*
chemical_list: D000906:Antibodies; D008051:Lipid Bilayers; D015222:Sodium Channels; D000450:Aldosterone