Biosynthesis of high molecular weight polylactosamine-type glycopeptides in rat Zajdela hepatoma ascites cells.
The first steps of the biosynthetic pathway of high molecular weight polylactosamine-type glycopeptides from rat Zajdela hepatoma cells were studied by pulse-chase experiments, biochemical analysis and by inhibition of N-glycosylation. It is clear that this process involves firstly the transfer of a lipid-linked high-mannose oligosaccharide precursor to a protein moiety in a similar way to that of N-linked glycopeptides of a more common size range according to the classical 'cycle of dolichol'. In the presence of enzymes which are inhibitors of the processing of high-mannose oligosaccharide chains, this class of oligosaccharides was considerably increased, whereas polylactosamine chains and lower complex N-linked glycopeptides were concomitantly decreased in the same kinetics and the same ratio. As expected in the presence of N-methyldeoxynojirimycin, which is an alpha-glucosidase inhibitor, high-mannose oligosaccharides remained glycosylated and are mostly of the Glc1-3Man9GlcNAc type. In the presence of swainsonine, which is an alpha-mannosidase (EC 3.2.1.24) inhibitor, these chains were devoid of glucose residues. In addition, some chains displayed hybrid structures. It appears, therefore, that the first steps of the biosynthesis of polylactosamine-type and N-linked oligosaccharides of a more common size range proceed similarly and that differences between their biosynthetic pathways occur during the elongation phase, which leads to their final respective structures. Glycopeptides prepared from the cell surface by mild trypsin treatment as well as from entire cells, previously treated or not by processing inhibitors, display the same gel filtration patterns indicating that modifications in protein glycosylation do not prevent glycoprotein insertion into the cell membrane.
Saunier
B
B
Laboratoire de Biochimie, Faculté de Médecine, Paris, France.
Goulut
C
C
Nato
F
F
Bourrillon
R
R
D000606:Amino Sugars / Q000096:biosynthesis; D000818:Animals; D001201:Ascites / Q000378:metabolism; D002845:Chromatography / Q000379:methods; D006020:Glycopeptides / Q000096:biosynthesis*; D008114:Liver Neoplasms, Experimental / Q000378:metabolism*; D008358:Mannose / Q000378:metabolism; D008970:Molecular Weight; D009844:Oligosaccharides / Q000378:metabolism; D051381:Rats; D014407:Tumor Cells, Cultured
D000606:Amino Sugars; D006020:Glycopeptides; D009844:Oligosaccharides; C032233:lactosamine; D008358:Mannose