Biosynthesis of a low-molecular-mass rat submandibular gland mucin glycoprotein in COS7 cells.
We have examined the biosynthesis of a low-molecular-mass mucin from rat submandibular gland (RSMG) expressed recombinantly in COS7 tissue culture cells, focusing primarily on the addition of carbohydrate to the protein core of the mucin. We find evidence for N-linked glycosylation, but this modification is not required for secretion of the mucin. Similarly, although the recombinant RSMG mucin, like its native counterpart, contains large amounts of O-linked carbohydrate, chain extension beyond the initial O-linked GalNAc moiety is not required for secretion. We have identified partially glycosylated mucin by a combination of metabolic pulse-chase and lectin precipitations of the biosynthetic intermediates. Our results suggest that the addition of GalNAc to threonine and serine in the RSMG mucin does not occur simultaneously, as has been described for other O-glycosylated proteins.
Nehrke
K
K
Department of Dental Research, School of Medicine and Dentistry, University of Rochester, 601 Elmwood Avenue, Box 611, Rochester, NY 14642, USA.
Tabak
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D000116:Acetylgalactosamine / Q000378:metabolism; D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D019556:COS Cells; D004591:Electrophoresis, Polyacrylamide Gel; D006026:Glycoside Hydrolases / Q000378:metabolism; D006031:Glycosylation; D008970:Molecular Weight; D009077:Mucins / Q000096:biosynthesis*; D051381:Rats; D011994:Recombinant Proteins / Q000096:biosynthesis; D012463:Saliva / Q000737:chemistry; D013363:Submandibular Gland / Q000378:metabolism*; D014415:Tunicamycin / Q000494:pharmacology
D000900:Anti-Bacterial Agents; D009077:Mucins; D011994:Recombinant Proteins; D014415:Tunicamycin; D006026:Glycoside Hydrolases; D000116:Acetylgalactosamine