The relationship between glycosylation and glycoprotein metabolism of mouse neuroblastoma N18 cells.
Two inhibitors of glycosylation, glucosamine and tunicamycin, were utilized to examine the effect of glycosylation inhibition in mouse neuroblastoma N18 cells on the degradation of membrane glycoproteins synthesized before addition of the inhibitor. Treatment with 10 mM-glucosamine resulted in inhibition of glycosylation after 2h, as measured by [3H]fucose incorporation into acid-insoluble macromolecules, and in a decreased rate of glycoprotein degradation. However, these results were difficult to interpret since glucosamine also significantly inhibited protein synthesis, which in itself could cause the alteration in glycoprotein degradation [Hudson & Johnson (1977) Biochim. Biophys. Acta 497, 567-577]. N18 cells treated with 5 microgram of tunicamycin/ml, a more specific inhibitor of glycosylation, showed a small decrease in protein synthesis relative to its effect on glycosylation, which was inhibited by 85%. Tunicamycin-treated cells also showed a marked decrease in glycoprotein degradation in experiments with intact cells. The inhibition of glycoprotein degradation by tunicamycin was shown to be independent of alterations in cyclic AMP concentration. Polyacrylamide-gel electrophoresis of isolated membranes from N18 cells, double-labelled with [14C]fucose and [3H]fucose, revealed heterogeneous turnover rates for specific plasma-membrane glycoproteins. Comparisons of polyacrylamide gels of isolated plasma membranes from [3H]fucose-labelled control cells and [14C]fucose-labelled tunicamycin-treated cells revealed that both rapidly and slowly metabolized, although not all, membrane glycoproteins became resistant to degradation after glycosylation inhibition.
D000818:Animals; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D003864:Depression, Chemical; D005643:Fucose / Q000378:metabolism; D005944:Glucosamine / Q000494:pharmacology; D006023:Glycoproteins / Q000378:metabolism*; D051379:Mice; D009447:Neuroblastoma / Q000378:metabolism* / Q000473:pathology; D014176:Protein Biosynthesis; D014415:Tunicamycin / Q000494:pharmacology
D006023:Glycoproteins; D014415:Tunicamycin; D005643:Fucose; D005944:Glucosamine