Purification of phospholipase C from rat cerebral cortex.
Phospholipase C from rat cerebral cortex was purified to homogeneity by use of DEAE Bio-Gel A agarose, hydroxyapatite, and heparin agarose chromatography. The purified phospholipase C (PLC) was purified 622.4-fold and its molecular weight is estimated to be 97,500. We obtained a final specific activity of 3.112 mumol of phosphatidylinositol hydrolyzed/min/mg of protein. It is specific for inositol phospholipids. The purified enzyme has an apparent optimum pH 7.0. Calcium is required for its activity. Western blotting analysis showed that two proteins were recognized by anti-PLC antiserum.
Wu
C Y
CY
Department of Pharmacology, National Yang-Ming Medical College, Taipei, Taiwan, Republic of China.
Chen
C F
CF
Chiang
C F
CF
D000818:Animals; D015153:Blotting, Western; D002118:Calcium / Q000494:pharmacology; D002540:Cerebral Cortex / Q000201:enzymology*; D002845:Chromatography; D006863:Hydrogen-Ion Concentration; D008970:Molecular Weight; D010716:Phosphatidylinositols / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D013379:Substrate Specificity; D010738:Type C Phospholipases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism
D010716:Phosphatidylinositols; D010738:Type C Phospholipases; D002118:Calcium