Biosynthesis and expression of the myelin-associated glycoprotein in cultured oligodendrocytes from adult bovine brain.
The biosynthesis and expression of myelin-associated glycoprotein (MAG) were investigated in cultured oligodendrocytes isolated from adult bovine brain. Western blotting revealed two prominent MAG bands that were present in comparable amounts; the larger component electrophoresed above the 97 kD standard but was slightly smaller than the MAG band in purified bovine myelin, and the smaller component electrophoresed below the 97 kD standard. In comparison to other precursors of oligosaccharides, inorganic [35S]sulfate was a relatively specific isotope for labeling MAG relative to other glycoproteins in the cells. Sulfate labeled only the larger of the two MAG components, which contains complex N-linked oligosaccharides, but which appears to be glycosylated to a lesser extent than MAG in vivo. The smaller MAG band in the cells is a form with high-mannose oligosaccharides and was not detected in purified bovine myelin. Both the large and small MAG components were expressed on the oligodendrocyte surface as indicated by their sensitivity to neuraminidase and/or trypsin treatment of live cells. MAG was also released by the oligodendrocytes into the culture medium. The MAG in the medium was slightly smaller than that in the cells, suggesting that it may be released from the cell surface by limited proteolysis. The release of MAG by myelin-forming cells could be relevant to physiological roles that have been postulated for soluble forms of MAG and other adhesion proteins.
D000818:Animals; D015153:Blotting, Western; D001923:Brain Chemistry / Q000502:physiology*; D002417:Cattle; D002478:Cells, Cultured; D003470:Culture Media; D006026:Glycoside Hydrolases / Q000494:pharmacology; D006868:Hydrolysis; D009185:Myelin Proteins / Q000096:biosynthesis* / Q000235:genetics; D018992:Myelin-Associated Glycoprotein; D009439:Neuraminidase / Q000494:pharmacology; D009836:Oligodendroglia / Q000201:enzymology / Q000378:metabolism*; D012756:Sheep; D014357:Trypsin / Q000494:pharmacology
D003470:Culture Media; D009185:Myelin Proteins; D018992:Myelin-Associated Glycoprotein; D006026:Glycoside Hydrolases; C020668:glycanase; D009439:Neuraminidase; D014357:Trypsin