Analysis of the phosphorylation state of a collagen-binding heat-shock glycoprotein from L6 myoblasts by isoelectric focusing.
A major collagen-binding glycoprotein from rat L6 skeletal myoblasts, designated gp46, is phosphorylated in vivo. In this report the relative phosphorylation state of gp46 was examined using isoelectric focusing to identify the phosphorylated and unphosphorylated forms of gp46. Two major and one minor isoform of gp46 were identified that could be related to the phosphorylation state of gp46. The relative percentage of unphosphorylated to phosphorylated gp46 increased 10% in myoblasts heat-shocked at 42 degrees C for 24 h. Treatment of myoblasts with phorbol ester or dibutyryl-cAMP had no effect on the phosphorylation ratio of gp46. Transformation of L6 myoblasts with Rous sarcoma virus, likewise, had no effect on the phosphorylation ratio. However, ras-transformed L6 myoblasts showed a 12% increase in phosphorylation of gp46. These results indicate that gp46 does not undergo large changes in phosphorylation status. Pulse-chase labelling showed that the phosphorylation of gp46 occurred either co-translationally or soon after translation, suggesting that gp46 was phosphorylated by a constitutively active protein kinase.
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Department of Biochemistry, Queen's University, Kingston, Canada.
D000818:Animals; D015153:Blotting, Western; D002460:Cell Line; D006360:Heat-Shock Proteins / Q000737:chemistry* / Q000302:isolation & purification; D007525:Isoelectric Focusing; D008562:Membrane Glycoproteins / Q000737:chemistry* / Q000302:isolation & purification; D009132:Muscles / Q000737:chemistry* / Q000166:cytology; D010766:Phosphorylation; D051381:Rats; D011956:Receptors, Cell Surface / Q000737:chemistry* / Q000302:isolation & purification; D038981:Receptors, Collagen
D006360:Heat-Shock Proteins; D008562:Membrane Glycoproteins; D011956:Receptors, Cell Surface; D038981:Receptors, Collagen