Oligosaccharyltransferase activity is markedly increased during differentiation of a nonfusing myoblast cell line.
We have studied several aspects of glycoprotein synthesis in myoblast differentiation by using a nonfusing myoblast cell line, BC3H1. Previous studies showed that transfer of proliferating undifferentiated BC3H1 cells to mitogen-depleted medium results in the cells' withdrawal from the cell cycle and induction of a variety of muscle-specific gene products [E. N. Olson, L. Glaser, J. P. Merlie, R. Sebane, and J. Lindstrom (1983) J. Biol. Chem. 258, 13946-13953]. Because cell surface glycoproteins have been implicated in myoblast differentiation, in the present study we measured the amount of oligosaccharyltransferase in microsomes isolated from BC3H1 cells at various stages of differentiation. By using an acceptor peptide containing the sequence-Asn-Leu-Thr-, enzyme activity was measured by formation of [3H]glycopeptide. In addition, active enzyme protein was measured with a 125I-labeled photoreactive derivative of the acceptor tripeptide. Both of these independent assay methods revealed a marked increase in oligosaccharyltransferase when differentiation was induced by serum depletion. Moreover, mitogenic stimulation of differentiated cells resulted in a return of oligosaccharyltransferase to near basal levels. This reversible increase in this key enzyme in protein glycosylation occurred despite the fact that both total protein and glycoprotein synthesis were depressed during differentiation. These data indicate that during myogenesis the level of oligosaccharyltransferase is regulated in parallel with a number of muscle-specific gene products. These results are discussed in the context of regulation of the pathway of glycoprotein synthesis.
D000345:Affinity Labels; D000818:Animals; D002454:Cell Differentiation; D002460:Cell Line; D004286:Dolichols / Q000378:metabolism; D006023:Glycoproteins / Q000096:biosynthesis*; D006602:Hexosyltransferases; D007700:Kinetics; D008358:Mannose / Q000378:metabolism; D008565:Membrane Proteins; D051379:Mice; D008861:Microsomes / Q000201:enzymology; D009132:Muscles / Q000166:cytology / Q000201:enzymology*; D010777:Photochemistry; D014166:Transferases / Q000378:metabolism*
D000345:Affinity Labels; D004286:Dolichols; D006023:Glycoproteins; D008565:Membrane Proteins; D014166:Transferases; D006602:Hexosyltransferases; C025620:dolichyl-diphosphooligosaccharide - protein glycotransferase; D008358:Mannose