Identification of lipid inhibitor of mammalian phospholipase D.
Phospholipase D (PLD) is implicated in important cellular processes, such as hormone action, inflammation, secretion, mitogenesis, and neural activity. Recent studies using cell-free systems have shown that the enzyme activity is modulated by both positive and negative regulators. During an attempt to purify PLD from pig colon mucosa, we noted the presence of a PLD inhibitor in the tissue extract. The inhibitor was purified and identified as comprising lysophosphatidylserine, phosphatidylinositol, and lysophosphatidylinositol, of which lysophosphatidylserine was the most potent. These lipids affected all of the PLD isoforms examined, oleate-dependent PLD, ARF-dependent PLD (PLD1a, PLD1b), and phosphatidylinositol 4,5-bisphosphate-dependent PLD (PLD2), in the concentration range of the 1 or 10 microM order. In contrast to lysophosphatidylserine, the diacyl counterpart phosphatidylserine was without effect in the same concentration range. PLD inhibition by lysophosphatidylserine could not be reversed by an increase in the concentration of the substrate phosphatidylcholine or activator phosphatidylinositol 4,5-bisphosphate.
Kawabe
K
K
Department of Biochemistry, Gunma University School of Medicine, Maebashi 371-8511, Japan.
Kodaki
T
T
Katayama
K
K
Okamura
S
S
Mori
M
M
Yamashita
S
S
D000818:Animals; D005591:Chemical Fractionation; D002850:Chromatography, Gel; D002855:Chromatography, Thin Layer; D003106:Colon / Q000737:chemistry / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*; D066298:In Vitro Techniques; D007413:Intestinal Mucosa / Q000737:chemistry / Q000378:metabolism; D007527:Isoenzymes / Q000037:antagonists & inhibitors*; D008246:Lysophospholipids / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology; D010716:Phosphatidylinositols / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology; D010718:Phosphatidylserines / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology; D010739:Phospholipase D / Q000037:antagonists & inhibitors*; D013552:Swine
D004791:Enzyme Inhibitors; D007527:Isoenzymes; D008246:Lysophospholipids; D010716:Phosphatidylinositols; D010718:Phosphatidylserines; C025449:lysophosphatidylinositol; C025059:lysophosphatidylserine; D010739:Phospholipase D