title: Studies on asparagine-linked protein glycosylation in differentiating skeletal muscle cells.
abstract: The embryonic development of skeletal muscle proceeds by the adherence and fusion of myoblast cells to form multinucleated myotubes. In the present study, enzymes in the dolichol pathway for asparagine-linked glycoprotein synthesis and oligosaccharide chain composition were characterized in myoblasts and myotubes derived from the C2 (mouse) muscle cell line. The N-acetylglucosaminyltransferase responsible for chain initiation and the mannosyl- and glucosyltransferases for Dol-P-Man and Dol-P-Glc synthesis were characterized with respect to substrate, cation, and detergent dependence. Time course studies in the absence and presence of exogenous Dol-P revealed that myoblasts had a two- to threefold higher capacity than myotubes for Dol-sugar synthesis. Pulse-chase experiments following the elongation of the Dol-oligosaccharide by intact cells showed myoblasts to label oligosaccharide intermediates approximately fourfold greater than myotubes; myotubes, however, were more efficient than myoblasts for converting the intermediates to the glucosylated Dol-tetradecasaccharide. Oligosaccharide chains isolated from sarcolemma glycopeptides were analyzed by Con A, WGA, and QAE chromatography. There were no differences between myoblast and myotube oligosaccharides with respect to the proportion of tri-tetraantennary complex, biantennary complex, and high mannose chains. Hybrid chains were not detected. The major high mannose chain contained nine mannose residues. Sialyltransferase activity was identical. The results suggest that higher levels of Dol-P and protein acceptor contribute to the greater degree of protein glycosylation in myoblast vs myotube muscle cells.
authors.affiliation: Department of Biochemistry, Bowman Gray School of Medicine, Wake Forest University, Winston-Salem, North Carolina 27103.
authors.forename: K R
authors.initials: KR
authors.lastname: Miller
authors.forename: C N
authors.initials: CN
authors.lastname: Remy
authors.forename: P B
authors.initials: PB
authors.lastname: Smith
mesh_terms: D000818:Animals; D002454:Cell Differentiation; D002460:Cell Line; D004286:Dolichols / Q000096:biosynthesis; D006023:Glycoproteins / Q000096:biosynthesis*; D006031:Glycosylation; D006602:Hexosyltransferases / Q000378:metabolism*; D007700:Kinetics; D051379:Mice; D024510:Muscle Development; D009132:Muscles / Q000378:metabolism*; D009844:Oligosaccharides / Q000096:biosynthesis
chemical_list: D004286:Dolichols; D006023:Glycoproteins; D009844:Oligosaccharides; D006602:Hexosyltransferases; C018660:asparagine-N-glycosyltransferase