Interaction of syntaxins with the amiloride-sensitive epithelial sodium channel.
Amiloride-sensitive sodium channels mediate sodium entry across the apical membrane of epithelial cells in variety of tissues. The rate of Na(+) entry is controlled by the regulation of the epithelial sodium channel (ENaC) complex. Insertion/retrieval of the ENaC complex into the apical membrane as well as direct kinetic effects at the single channel level are recognized mechanisms of regulation. Recent data suggest that the syntaxin family of targeting proteins interact with and functionally regulate a number of ion channels and pumps. To evaluate the role of these proteins in regulating ENaC activity, we co-expressed rat ENaC cRNA (alpha, beta, gamma subunits) with syntaxin 1A or 3 cRNAs in Xenopus oocytes. Basal ENaC currents were inhibited by syntaxin 1A and stimulated by syntaxin 3. Both syntaxin 1A and syntaxin 3 could be co-immunoprecipitated with ENaC subunit proteins, suggesting physical interaction. Interestingly, immunofluorescence data suggest that with either syntaxin isoform the ENaC-associated epifluorescence on the oocyte surface is enhanced. These data indicate that (i) both syntaxin isoforms increase the net externalization of the ENaC channel complex, (ii) that the functional regulation is isoform specific, and (iii) suggest that ENaC may be regulated through mechanisms involving protein-protein interactions.
Saxena
S
S
Nephrology Research and Training Center, University of Alabama at Birmingham, Birmingham, Alabama 35294, USA.
Quick
M W
MW
Tousson
A
A
Oh
Y
Y
Warnock
D G
DG
D000818:Animals; D001692:Biological Transport; D053503:Epithelial Sodium Channels; D005786:Gene Expression Regulation; D006801:Humans; D015640:Ion Channel Gating; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism; D050765:Qa-SNARE Proteins; D051381:Rats; D015222:Sodium Channels / Q000235:genetics / Q000378:metabolism*; D050827:Syntaxin 1; D014981:Xenopus
D053503:Epithelial Sodium Channels; D008565:Membrane Proteins; D020033:Protein Isoforms; D050765:Qa-SNARE Proteins; C494212:STX1A protein, human; D015222:Sodium Channels; C494213:Stx1a protein, rat; D050827:Syntaxin 1