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Purification of phosphatidylinositol-specific phospholipase C from NG108-15 cells
C F Chiang1, W L Chai, C F Chen
1Department of Pharmacology, National Yang-Ming Medical College, Taipei, Taiwan, Republic of China.
Abstract:
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.