Biosynthesis, processing and secretion of mucus glycoprotein in the rat stomach.
For the study of the biosynthesis, processing and secretion of mucus glycoproteins in rat gastric mucous cells, antibodies were raised against purified gastric mucus glycoproteins and against deglycosylated gastric mucus glycoproteins. Indirect immunofluorescence analysis of gastric mucosa sections revealed that both antibodies specifically labelled the mucus glycoprotein-synthesizing cells in the gastric mucosa. Stomach segments were pulse-labelled with [35S]cysteine and chased for various times. The radioactively labelled (glyco)proteins were quantitatively immunoprecipitated and analyzed by SDS-polyacrylamide gel electrophoresis. Less than 3% of the total radioactivity incorporated in protein was found to be present in mucus glycoproteins. Antibodies raised against native mucus glycoproteins recognized only high-molecular-weight mucus glycoproteins, while the antibodies against deglycosylated glycoproteins also bound to probable precursor forms. The synthesis of mature mucus glycoproteins (Mr greater than 300 000) required about 90 min. After 3 h of chase, only a small portion of the pulse-labelled mucus glycoproteins had been secreted; the majority of the radioactive glycoproteins at that time was still associated with the tissue. Immature (glyco)proteins were not secreted into the medium.
D000818:Animals; D011232:Chemical Precipitation; D005455:Fluorescent Antibody Technique; D005753:Gastric Mucosa / Q000378:metabolism*; D006023:Glycoproteins / Q000096:biosynthesis*; D007163:Immunosorbent Techniques; D008297:Male; D008970:Molecular Weight; D009093:Mucus / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D051381:Rats; D013997:Time Factors
D006023:Glycoproteins