| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
J. Biol. Chem., Vol. 265, Issue 29, 17512-17519, 10, 1990
V Chalifa, H Mohn and M Liscovitch
Department of Hormone Research, Weizmann Institute of Science, Rehovot, Israel.
Rapid activation of phospholipase D (PLD) in response to cell stimulation was recently demonstrated in many systems, raising the hypothesis that PLD participates in transduction of extracellular signals across the plasma membrane. In the present study, we describe the identification of a neutral PLD activity in purified rat brain synaptic plasma membranes, and the in vitro conditions required to assay its catalytic activity with exogenous [3H]phosphatidylcholine as substrate. Production of [3H]phosphatidic acid, the natural lipid product of PLD and of [3H]phosphatidylethanol, catalyzed by PLD in the presence of ethanol via transphosphatidylation, were measured. The synaptic membrane PLD exhibited its highest activity at pH 7.2 and was thus defined as a neutral PLD. Enzyme activity was absolutely dependent on the presence of sodium oleate and was strongly activated by Mg2+ ions (at 1 mM). Ca2+ at concentrations up to 0.25 mM was as stimulatory as Mg2+, but at 2 mM it completely inhibited enzyme activity. Mg2+ extended the linear phase of PLD activity from 2 to 15 min, suggesting that it may stabilize the enzyme under our assay conditions. The production of [3H]phosphatidylethanol was a saturable function of ethanol concentration. Production of [3H] phosphatidic acid was inversely related to the concentration of ethanol and to the accumulation of phosphatidylethanol, indicating that the two phospholipids are indeed produced by the competing hydrolase and transferase activities of the same enzyme. beta,beta-Dimethylglutaric acid, utilized previously as a buffer in studies of rat brain PLD, inhibited enzyme activity at neutral pH but not at acidic pH. The properties of the neutral synaptic membrane PLD and its relationships with other in vitro, acid, and neutral PLD activities, as well as with the signal-dependent PLD detected in intact cells, are discussed.
CiteULike 
Complore 
Connotea 
Del.icio.us 
Digg 
Reddit 
Technorati What's this?
This article has been cited by other articles:
|
|
 |
|
  D. Duijsings, E. Wessels, S. E. van Emst-de Vries, W. J. G. Melchers, P. H. G. M. Willems, and F. J. M. van Kuppeveld Reduction of phospholipase D activity during coxsackievirus infection J. Gen. Virol., November 1, 2007; 88(11): 3027 - 3030. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 O. Diaz, A. Berquand, M. Dubois, S. Di Agostino, C. Sette, S. Bourgoin, M. Lagarde, G. Nemoz, and A.-F. Prigent The Mechanism of Docosahexaenoic Acid-induced Phospholipase D Activation in Human Lymphocytes Involves Exclusion of the Enzyme from Lipid Rafts J. Biol. Chem., October 11, 2002; 277(42): 39368 - 39378. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. H. Kim, B. D. Lee, Y. Kim, S. D. Lee, P.-G. Suh, and S. H. Ryu Cytosolic Phospholipase A2-Mediated Regulation of Phospholipase D2 in Leukocyte Cell Lines J. Immunol., November 15, 1999; 163(10): 5462 - 5470. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. S. Arneson, J. Kunz, R. A. Anderson, and L. M. Traub Coupled Inositide Phosphorylation and Phospholipase D Activation Initiates Clathrin-coat Assembly on Lysosomes J. Biol. Chem., June 18, 1999; 274(25): 17794 - 17805. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S.-i. Nakamura, T. Akisue, H. Jinnai, T. Hitomi, S. Sarkar, N. Miwa, T. Okada, K. Yoshida, S.'i. Kuroda, U. Kikkawa, et al. Requirement of GM2 ganglioside activator for phospholipase D activation PNAS, October 13, 1998; 95(21): 12249 - 12253. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Bechoua, M. Dubois, G. Némoz, M. Lagarde, and A.-F. Prigent Docosahexaenoic acid lowers phosphatidate level in human activated lymphocytes despite phospholipase D activation J. Lipid Res., April 1, 1998; 39(4): 873 - 883. [Abstract] [Full Text]
|
|
|
|
|
|
Y. Banno, K. Tamiya-Koizumi, H. Oshima, A. Morikawa, S. Yoshida, and Y. Nozawa Nuclear ADP-ribosylation Factor (ARF)- and Oleate-dependent Phospholipase D (PLD) in Rat Liver Cells. INCREASES OF ARF-DEPENDENT PLD ACTIVITY IN REGENERATING LIVER CELLS J. Biol. Chem., February 21, 1997; 272(8): 5208 - 5213. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J.-S. Han, J.-K. Chung, H.-S. Kang, J. Donaldson, Y. S. Bae, and S. G. Rhee Multiple Forms of Phospholipase D Inhibitor from Rat Brain Cytosol J. Biol. Chem., May 10, 1996; 271(19): 11163 - 11169. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Y Nishizuka Intracellular signaling by hydrolysis of phospholipids and activation of protein kinase C Science, October 23, 1992; 258(5082): 607 - 614. [Abstract] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
| All ASBMB Journals | Molecular and Cellular Proteomics |
| Journal of Lipid Research | ASBMB Today |
