title: Novel function of phosphatidylinositol 4,5-bisphosphate as a cofactor for brain membrane phospholipase D.
abstract: The activation of phospholipase D (PLD) is a receptor-mediated event that has been implicated in signal transduction and membrane traffic in eukaryotic cells. Little is known about the biochemical and molecular properties of signal-activated PLDs, and none has been isolated. Here we report that phosphatidylinositol 4,5-bisphosphate (PIP2) potently stimulates brain membrane PLD activity in vitro in a highly specific manner. PIP2 increases 10-fold the maximal activity of a partially purified PLD with an EC50 of < 0.5 mol %. Other acidic phospholipids, including phosphatidylinositol 4-phosphate, phosphatidylinositol, phosphatidylserine, and phosphatidic acid, are completely or nearly ineffective. Neomycin, a high affinity ligand of PIP2, inhibits membrane-bound PLD but has no effect on the activity of a detergent-solubilized or partially purified enzyme. The addition of PIP2 restores the sensitivity of partially purified PLD to neomycin inhibition, indicating that neomycin blocks membrane PLD activity by binding to endogenous PIP2. These results define a novel function of PIP2 as a cofactor for brain membrane PLD and suggest that PIP2 synthesis and hydrolysis could be important determinants in regulating PLD action in signal transduction and membrane transport.
authors.affiliation: Department of Hormone Research, Weizmann Institute of Science, Rehovot, Israel.
authors.forename: M
authors.initials: M
authors.lastname: Liscovitch
authors.forename: V
authors.initials: V
authors.lastname: Chalifa
authors.forename: P
authors.initials: P
authors.lastname: Pertile
authors.forename: C S
authors.initials: CS
authors.lastname: Chen
authors.forename: L C
authors.initials: LC
authors.lastname: Cantley
mesh_terms: D000818:Animals; D001921:Brain / Q000201:enzymology*; D004789:Enzyme Activation; D008566:Membranes / Q000201:enzymology; D008954:Models, Biological; D009355:Neomycin / Q000494:pharmacology; D019269:Phosphatidylinositol 4,5-Diphosphate; D018129:Phosphatidylinositol Phosphates / Q000378:metabolism / Q000494:pharmacology*; D010739:Phospholipase D / Q000187:drug effects / Q000378:metabolism*; D051381:Rats
chemical_list: D019269:Phosphatidylinositol 4,5-Diphosphate; D018129:Phosphatidylinositol Phosphates; C060974:phosphatidylinositol 3,4,5-triphosphate; D010739:Phospholipase D; D009355:Neomycin