Partial purification of phosphoinositide phospholipase C from human platelet cytosol; characterization of its three forms

Biochem Biophys Res Commun. 1986 Apr 29;136(2):713-21. doi: 10.1016/0006-291x(86)90498-5.

Abstract

Three forms (I, IIa and IIb) of phospholipase C, hydrolyzing specifically inositol phospholipids, were resolved from human platelet cytosol and partially purified by DEAE-cellulose, phenyl-Sepharose, Ultrogel ACA-44 and hydroxylapatite column chromatographies. All three forms exhibited pH optimum at 6.5 - 7.0 in the presence of deoxycholate and their molecular weights were 67,000 (form I), 120,000 (IIa) and 70,000 (IIb). These enzymes hydrolyzed both phosphatidylinositol and phosphatidylinositol 4,5-bisphosphate in Ca2+-dependent manner; their maximal activities for phosphatidylinositol hydrolysis were obtained at 10(-4) to 10(-3) M Ca2+, whereas phosphatidylinositol 4,5-bisphosphate was preferentially hydrolyzed at lower concentration of Ca2+.

Publication types

  • Comparative Study

MeSH terms

  • Blood Platelets / enzymology*
  • Calcium / pharmacology
  • Chromatography
  • Cytosol / enzymology
  • Humans
  • Hydrogen-Ion Concentration
  • Hydrolysis
  • Kinetics
  • Molecular Weight
  • Phosphatidylinositol 4,5-Diphosphate
  • Phosphatidylinositol Diacylglycerol-Lyase
  • Phosphatidylinositols / metabolism
  • Phosphoric Diester Hydrolases / blood*

Substances

  • Phosphatidylinositol 4,5-Diphosphate
  • Phosphatidylinositols
  • Phosphoric Diester Hydrolases
  • Phosphatidylinositol Diacylglycerol-Lyase
  • Calcium