The voltage-sensitive sodium channel from rabbit skeletal muscle. Chemical characterization of subunits.
The alpha and beta subunits of the rabbit skeletal muscle sodium channel have been separated and isolated preparatively under denaturing conditions. In this sodium channel, the beta subunit is not linked covalently to the alpha subunit. The isolated subunits have been subjected to amino acid and carbohydrate analysis. Both subunits are heavily glycosylated (alpha = 26.5%, beta = 29.7% carbohydrate by weight) with N-acetylneuraminic acid and N-acetylhexosamines representing the predominant monosaccharides in each. Enzymatic deglycosylation with neuraminidase and endoglycosidase F yielded single core peptides of approximately 209 kDa for the alpha subunit and 26.5 kDa for the beta subunit. Based on the known carbohydrate composition, the molecular masses for the glycosylated subunits are, therefore, 285 and 37.5 kDa for alpha and beta, respectively. Using the isolated subunits, we calibrated our protein-labeling system on sodium dodecyl sulfate-polyacrylamide gel electrophoresis and determined the subunit stoichiometry for the rabbit skeletal muscle channel; in the native preparation, the molar ratio of alpha:beta is 1 : 1.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002241:Carbohydrates / Q000032:analysis; D006026:Glycoside Hydrolases / Q000378:metabolism; D007473:Ion Channels / Q000032:analysis / Q000378:metabolism*; D046911:Macromolecular Substances; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase; D008970:Molecular Weight; D009132:Muscles / Q000032:analysis*; D019158:N-Acetylneuraminic Acid; D009439:Neuraminidase / Q000378:metabolism; D011489:Protein Denaturation; D011817:Rabbits; D012794:Sialic Acids / Q000032:analysis; D012964:Sodium / Q000378:metabolism*
D000596:Amino Acids; D002241:Carbohydrates; D007473:Ion Channels; D046911:Macromolecular Substances; D012794:Sialic Acids; D012964:Sodium; D006026:Glycoside Hydrolases; D009439:Neuraminidase; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase; D019158:N-Acetylneuraminic Acid