Phospholipase C from human melanoma: purification and characterization of a phosphatidylinositol-selective enzyme.
Phospholipase C was purified from human melanoma grown as solid tumors in nude mice. The specific activity of the pure enzyme was approx. 100 mumol/min per mg; its apparent molecular mass was determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis to be 150 kDa. The enzyme required calcium for activity and was activated by deoxycholate in the presence of the substrate phosphatidylinositol. The melanoma phospholipase C has a distinctly different substrate preference than those identified from normal tissues; it prefers phosphatidylinositol to phosphatidylinositol bisphosphate. The tumor enzyme was approx. 4-5-fold more active using phosphatidylinositol than phosphatidylinositol bisphosphate as the substrate.
Perrella
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Medical Products Department, E.I. DuPont de Nemours and Company, Glenolden, PA 19036.
Jankewicz
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Dandrow
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EA
D000818:Animals; D002460:Cell Line; D002845:Chromatography / Q000379:methods; D002846:Chromatography, Affinity / Q000379:methods; D002852:Chromatography, Ion Exchange / Q000379:methods; D017886:Durapatite; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D006882:Hydroxyapatites; D007700:Kinetics; D008545:Melanoma / Q000201:enzymology*; D051379:Mice; D008819:Mice, Nude; D008970:Molecular Weight; D009368:Neoplasm Transplantation; D010716:Phosphatidylinositols / Q000378:metabolism*; D013379:Substrate Specificity; D014183:Transplantation, Heterologous; D010738:Type C Phospholipases / Q000302:isolation & purification* / Q000378:metabolism
D006882:Hydroxyapatites; D010716:Phosphatidylinositols; D017886:Durapatite; D010738:Type C Phospholipases