Effects of tunicamycin, N-methyl-1-deoxynojirimycin, and manno-1-deoxynojirimycin on the biosynthesis of lactosaminoglycans in F9 teratocarcinoma cells.

F9 teratocarcinoma cells were incubated with D-[2-3H]mannose or D-[6-3H]galactose, and the labeled glycopeptides obtained after exhaustive digestion by pronase were fractionated on Bio-Gel P-6 before and after treatment by endo-beta-N-acetylglucosaminidase H. Tunicamycin almost completely inhibited the synthesis of lactosaminoglycans found in excluded glycopeptides of large molecular weight. Manno-1-deoxynojirimycin greatly inhibited the incorporation of labeled mannose into both lactosaminoglycan and complex oligosaccharides, while it greatly increased that into Man8GlcNAc and Man9GlcNAc oligosaccharides. In contrast, N-methyl-1-deoxynojirimycin only partially inhibited the incorporation into lactosaminoglycan and complex oligosaccharides, and caused the accumulation of Glc3Man7-9GlcNAc oligosaccharides. These results demonstrate that, in these cells, lactosaminoglycans are N-linked, and suggest that there is transfer of both glucosylated and nonglucosylated oligosaccharides from lipid to protein.