Partial characterization of a collagen-binding, differentiation-related glycoprotein from skeletal myoblasts.
A 46-kDa glycoprotein, gp46, which binds collagen has been purified to homogeneity from L6 rat skeletal myoblasts. The procedure involves extraction of crude myoblast membranes with 1% sodium dodecyl sulfate followed by concanavalin A affinity chromatography and preparative gel electrophoresis. The sequence of 15 N-terminal amino acids had some resemblance to a sequence in myosin light chains. The oligosaccharide chains of the glycoprotein can be released by treatment with endoglycosidase H, suggesting that gp46 has high-mannose type of glycans. Galactose and sialic acid are not detected in the purified protein. gp46 is widely distributed and conserved in different cell lines as determined by immunoblotting using a monoclonal anti-gp46 antibody. High levels of gp46 were found in several fibroblastic and myogenic cell lines, but not in a hematopoietic cell line. Undifferentiated F9 embryonal carcinoma cells lacked gp46 but the glycoprotein was induced when the cells were made to differentiate in the presence of retinoic acid. Broad survey of gp46 in different cell lines also suggests that it is present mainly in those cell lines which attach to the substratum and produce collagens. Although the function of gp46 is not yet known, the evidence suggests that it is developmentally regulated and is probably involved in the synthesis or assembly of collagen in the endoplasmic reticulum.
Nandan
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Department of Biochemistry, University of Western Ontario, London, Canada.
Cates
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GA
Ball
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Sanwal
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BD
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002454:Cell Differentiation; D003208:Concanavalin A / Q000378:metabolism; D006023:Glycoproteins / Q000302:isolation & purification*; D006801:Humans; D051379:Mice; D009132:Muscles / Q000032:analysis*; D009928:Organ Specificity; D011956:Receptors, Cell Surface / Q000302:isolation & purification*; D038981:Receptors, Collagen; D012995:Solubility; D014407:Tumor Cells, Cultured
D000596:Amino Acids; D006023:Glycoproteins; D011956:Receptors, Cell Surface; D038981:Receptors, Collagen; D003208:Concanavalin A