Biosynthesis and maturation of cellular membrane glycoproteins.
The biosynthesis and the processing of asparagine-linked oligosaccharides of cellular membrane glycoproteins were examined in monolayer cultures of BHK21 cells and human diploid fibroblasts after pulse- and pulse-chase labeling with [2-3H]mannose. After pronase digestion, radiolabeled glycopeptides were characterized by high-resolution gel filtration, with or without additional digestion with various exoglycosidases and endoglycosidases. Pulse-labeled glycoproteins contained a relatively homogenous population of neutral oligosaccharides (major species: Man9GlcNAc2ASN). The vast majority of these asparagine-linked oligosaccharides was smaller than the major fraction of lipid-linked oligosaccharides from the cell and was apparently devoid of terminal glucose. After pulse-chase or long labeling periods, a significant fraction of the large oligomannosyl cores was processed by removal of mannose units and addition of branch sugars (NeuNAc-Gal-GlcNAc), resulting in complex acidic structures containing three and possibly five mannoses. In addition, some of the large oligomannosyl cores were processed by the removal of only several mannoses, resulting in a mixture of neutral structures with 5-9 mannoses. This oligomannosyl core heterogeneity in both neutral and acidic oligosaccharides linked to asparagine in cellular membrane glycoproteins was analogous to the heterogeneity reported for the oligosaccharides of avian RNA tumor virus glycoproteins (Hunt LA, Wright SE, Etchison JR, Summers DF: J Virol 29:336, 1979).
D000118:Acetylglucosaminidase; D000818:Animals; D001216:Asparagine / Q000378:metabolism; D002240:Carbohydrate Sequence; D002460:Cell Line; D006224:Cricetinae; D004622:Embryo, Mammalian; D005347:Fibroblasts / Q000378:metabolism; D006020:Glycopeptides / Q000032:analysis; D006023:Glycoproteins / Q000096:biosynthesis*; D006367:HeLa Cells / Q000378:metabolism; D006801:Humans; D007668:Kidney; D008168:Lung; D008565:Membrane Proteins / Q000096:biosynthesis*; D009844:Oligosaccharides / Q000378:metabolism
D006020:Glycopeptides; D006023:Glycoproteins; D008565:Membrane Proteins; D009844:Oligosaccharides; D001216:Asparagine; D000118:Acetylglucosaminidase