Open AccessPurification of phosphoinositide‐specific phospholipase C from a particulate fraction of bovine brain
Author(s) -
KATAN Matilda,
PARKER Peter J.
Publication year - 1987
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1987.tb13435.x
Subject(s) - phospholipase c , chemistry , phosphatidylinositol , biochemistry , enzyme , chromatography , gel permeation chromatography , phospholipase , phosphoinositide phospholipase c , context (archaeology) , biology , signal transduction , organic chemistry , paleontology , polymer
The coupling of various agonist receptors to the hydrolysis of phosphoinositides has generated much interest in the nature of the phospholipase C that is activated. Here we report the purification of a bovine brain phospholipase C derived from the particulate fraction. A 1000‐fold purification was achieved by a combination of heparin‐Sepharose, DEAE‐cellulose and gel‐permeation chromatography. The purified enzyme appears to be monomeric and under denaturing conditions shows a single staining major polypeptide of molecular mass 154 kDa in SDS gels. The enzyme is specific for phosphoinositides although it shows a marked preference for the polyphosphoinositides. With phosphatidylinositol 4,5‐bisphosphate as substrate the enzyme expresses a specific activity of > 100 μmol min −1 mg −1 . The phospholipase C is activated by Ca 2+ (0.1 – 10 μM). The behaviour of this particulate enzyme is discussed in the context of agonist‐induced phosphatidylinositol hydrolysis.