Asparagine-linked oligosaccharides of BHK cells treated with inhibitors of oligosaccharide processing.
Baby-hamster kidney (BHK) cells were labelled with [2-3H]mannose for 1-2 days in media containing 1-deoxynojirimycin, N-methyl-1-deoxynojirimycin or 1-deoxymannojirimycin. Glycopeptides obtained by Pronase digestion of disrupted cells were analysed by lectin affinity chromatography, by Bio-Gel P4 gel filtration and by paper chromatography of oligosaccharides released by endo-beta-N-acetylglucosaminidase H. Biosynthesis of complex-type oligosaccharides was diminished but not abolished, the greatest effect being obtained by continuous culture of cells with 1-deoxymannojirimycin. Under these conditions cells contained only 20-30% of the concentration of complex-type chains found in control cells and correspondingly increased amounts of oligomannose-type chains. Similar concentrations of asparagine-linked Man6-GlcNAc2 and Man5GlcNAc2 were present in 1-deoxymannojirimycin-treated cells and control cells, indicating that the inhibition of complex-type chain formation was not related simply to an inability of inhibitor-treated cells to carry out extensive mannosidase-catalysed processing. N-Methyl-1-deoxynojirimycin induced accumulation of oligomannose-type chains containing three glucose residues, and cells treated with 1-deoxynojirimycin contained oligosaccharides with one to three glucose residues. Cells cultured in the presence of the inhibitors retained sensitivity towards the galactose-binding lectins ricin and modeccin.
D017485:1-Deoxynojirimycin / Q000031:analogs & derivatives; D000818:Animals; D001216:Asparagine; D002460:Cell Line; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D006224:Cricetinae; D005944:Glucosamine / Q000031:analogs & derivatives* / Q000494:pharmacology; D005947:Glucose / Q000494:pharmacology; D006020:Glycopeptides / Q000032:analysis; D006031:Glycosylation; D037102:Lectins / Q000494:pharmacology; D008358:Mannose / Q000378:metabolism; D008361:Mannosidases / Q000494:pharmacology; D009844:Oligosaccharides / Q000037:antagonists & inhibitors* / Q000378:metabolism; D043323:alpha-Mannosidase
D006020:Glycopeptides; D037102:Lectins; D009844:Oligosaccharides; D017485:1-Deoxynojirimycin; D001216:Asparagine; D008361:Mannosidases; D043323:alpha-Mannosidase; D005947:Glucose; D005944:Glucosamine; D008358:Mannose; C033424:N-methyldeoxynojirimycin