Purification of phosphatidylinositol-specific phospholipase C from NG108-15 cells.
The phosphatidylinositol-specific phospholipase C from the membrane of NG108-15 cells has been purified to homogeneity by using DEAE Bio-Gel A agarose, hydroxyapatite, and heparin agarose chromatography. The purified phosphatidylinositol-specific phospholipase C has been purified 422-fold and its molecular weight has been estimated to be 28,000. We have obtained a final specific activity of 3.8 mumols of phosphatidylinositol hydrolyzed/min/mg of protein. The purified enzyme is specific to phosphatidylinositol. The purified enzyme has an apparent pH optimum of 7.0. Calcium ions are required for its activity.
Chiang
C F
CF
Department of Pharmacology, National Yang-Ming Medical College, Taipei, Taiwan, Republic of China.
Chai
W L
WL
Chen
C F
CF
D000818:Animals; D002118:Calcium / Q000502:physiology; D002462:Cell Membrane / Q000201:enzymology; D002845:Chromatography; D004591:Electrophoresis, Polyacrylamide Gel; D005910:Glioma; D006863:Hydrogen-Ion Concentration; D051379:Mice; D008970:Molecular Weight; D009447:Neuroblastoma; D043265:Phosphatidylinositol Diacylglycerol-Lyase; D054801:Phosphoinositide Phospholipase C; D010727:Phosphoric Diester Hydrolases / Q000302:isolation & purification*; D051381:Rats; D014407:Tumor Cells, Cultured
D010727:Phosphoric Diester Hydrolases; D054801:Phosphoinositide Phospholipase C; D043265:Phosphatidylinositol Diacylglycerol-Lyase; D002118:Calcium