Cell surface expression and biosynthesis of epithelial Na+ channels.
The epithelial Na+ channel (ENaC) complex is composed of three homologous subunits: alpha, beta and gamma. Mutations in ENaC subunits can increase the number of channels on the cell surface, causing a hereditary form of hypertension called Liddle's syndrome, or can decrease channel activity, causing pseudohypoaldosteronism type I, a salt-wasting disease of infancy. To investigate surface expression, we studied ENaC subunits expressed in COS-7 and HEK293 cells. Using surface biotinylation and protease sensitivity, we found that when individual ENaC subunits are expressed alone, they traffic to the cell surface. The subunits are glycosylated with high-mannose oligosaccharides, but seem to have the carbohydrate removed before they reach the cell surface. Moreover, subunits form a complex that cannot be disrupted by several non-ionic detergents. The pattern of glycosylation and detergent solubility/insolubility persists when the N-teminal and C-terminal cytoplasmic regions of ENaC are removed. With co-expression of all three ENaC subunits, the insoluble complex is the predominant species. These results show that ENaC and its family members are unique in their trafficking, biochemical characteristics and post-translational modifications.
Prince
L S
LS
Howard Hughes Medical Institute, Departments of Pediatrics, Internal Medicine, and Physiology and Biophysics, University of Iowa College of Medicine, Iowa City, IA 52242, USA.
Welsh
M J
MJ
D000818:Animals; D020126:Brefeldin A / Q000494:pharmacology; D019556:COS Cells; D053503:Epithelial Sodium Channels; D006031:Glycosylation; D006801:Humans; D016296:Mutagenesis; D011487:Protein Conformation; D015222:Sodium Channels / Q000096:biosynthesis*; D012995:Solubility; D013499:Surface Properties; D014415:Tunicamycin / Q000494:pharmacology
D053503:Epithelial Sodium Channels; D015222:Sodium Channels; D014415:Tunicamycin; D020126:Brefeldin A