Translocation of phospholipase A2 to membranes by oxidized LDL and hydroxyoctadecadienoic acid to contribute to cholesteryl ester formation.
We examined the mechanisms underlying the activation of group IVA cytosolic phospholipase A(2) (cPLA(2)alpha) contributing to the supply of fatty acids required for the formation of cholesteryl ester in oxidized low-density lipoprotein (oxLDL)-stimulated macrophages. The possible involvement of oxidized lipids was also examined. In [(3)H]arachidonic acid-labeled mouse peritoneal macrophages, oxLDL stimulated the release of arachidonic acid, which was suppressed by methyl arachidonyl fluorophosphonate (MAFP), a cPLA(2)alpha inhibitor. oxLDL induced an increase in PLA(2)alpha levels in the membrane fraction without affecting those in whole cells or the activity in the lysate. Among 13-hydroxyoctadecadienoic acid (13-HODE), 7-ketocholesterol, and 25-hydroxycholesterol, oxidized lipids present in oxLDL particles, only 13-HODE induced the release of arachidonic acid, which was also sensitive to MAFP. Under conditions where addition of Ca(2+) to the cell lysate induced an increase in cPLA(2)alpha protein in the membrane fraction, preincubation with 13-HODE facilitated the Ca(2+)-dependent translocation of cPLA(2)alpha. Furthermore, 13-HODE increased cholesteryl ester formation in the presence of [(3)H]cholesterol. These results suggest that 13-HODE mediates the oxLDL-induced activation of cPLA(2)alpha through an increase in cPLA(2)alpha protein in the membranes, thus contributing, in part, to the supply of fatty acids required for the esterification of cholesterol in macrophages.
Akiba
Satoshi
S
Department of Pathological Biochemistry, Kyoto Pharmaceutical University, 5 Nakauchi-cho, Misasagi, Yamashina-ku, Kyoto 607-8414, Japan.
Ii
Hiromi
H
Yoneda
Yukimasa
Y
Sato
Takashi
T
D000818:Animals; D016718:Arachidonic Acid / Q000037:antagonists & inhibitors / Q000096:biosynthesis; D001095:Arachidonic Acids / Q000494:pharmacology; D000001:Calcimycin / Q000494:pharmacology; D002118:Calcium / Q000737:chemistry / Q000378:metabolism; D002462:Cell Membrane / Q000201:enzymology; D002478:Cells, Cultured; D002784:Cholesterol / Q000737:chemistry / Q000378:metabolism; D002788:Cholesterol Esters / Q000096:biosynthesis*; D003600:Cytosol / Q000201:enzymology; D004789:Enzyme Activation / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005260:Female; D006801:Humans; D007476:Ionophores / Q000494:pharmacology; D008041:Linoleic Acids / Q000378:metabolism / Q000494:pharmacology*; D008077:Lipoproteins, LDL / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D017737:Macrophages, Peritoneal / Q000187:drug effects / Q000378:metabolism; D051379:Mice; D063065:Organophosphonates / Q000494:pharmacology; D010084:Oxidation-Reduction; D010741:Phospholipases A / Q000037:antagonists & inhibitors / Q000378:metabolism*; D054467:Phospholipases A2; D021381:Protein Transport
D001095:Arachidonic Acids; D002788:Cholesterol Esters; D004791:Enzyme Inhibitors; D007476:Ionophores; D008041:Linoleic Acids; D008077:Lipoproteins, LDL; D063065:Organophosphonates; C104810:methyl arachidonylfluorophosphonate; D016718:Arachidonic Acid; D000001:Calcimycin; C024348:13-hydroxy-9,11-octadecadienoic acid; D002784:Cholesterol; D010741:Phospholipases A; D054467:Phospholipases A2; D002118:Calcium