Biosynthesis of gastric mucus glycoprotein of the rat.
We have studied the biosynthesis of rat gastric mucin in stomach segments using an antiserum against rat gastric mucin specific for peptide epitopes. Pulse-chase experiments were performed with [35S]methionine, [3H]galactose, and [35S]sulfate to label mucin precursors in different stages of biosynthesis, which were analyzed after immunoprecipitation. The earliest mucin precursor that could be detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis was a 300-kDa protein. The occurrence of N-linked "high-mannose" oligosaccharides on this protein was shown by susceptibility to degradation by endo-beta-N-acetylglucosaminidase H. This precursor could be labeled with [35S]methionine and not with [3H]galactose or [35S]sulfate. The 300-kDa precursor was converted into mature mucin after extensive glycosylation and sulfation. The mature mucin but not the 300-kDa precursor was in part secreted into the medium. Specific inhibition of sulfation with sodium chlorate had no effect on rate and amount of mucin secretion. In addition, we show that two core proteins are expressed in rats, slightly varying in Mr among individual animals.
Dekker
J
J
Department of Cell Biology, State University of Utrecht, School of Medicine, The Netherlands.
Van Beurden-Lamers
W M
WM
Strous
G J
GJ
D000818:Animals; D005690:Galactose / Q000378:metabolism; D005753:Gastric Mucosa / Q000378:metabolism*; D006023:Glycoproteins / Q000096:biosynthesis* / Q000302:isolation & purification; D006031:Glycosylation; D007700:Kinetics; D008297:Male; D008715:Methionine / Q000378:metabolism; D008970:Molecular Weight; D009077:Mucins / Q000302:isolation & purification; D051381:Rats; D011919:Rats, Inbred Strains; D013462:Sulfur Radioisotopes; D014316:Tritium
D006023:Glycoproteins; D009077:Mucins; D013462:Sulfur Radioisotopes; C047874:gastric mucus glycoproteins; D014316:Tritium; D008715:Methionine; D005690:Galactose