In the present study the lysophospholipid sources for arachidonic (AA) and eicosapentaenoic acid (EPA) incorporation into and redistribution within the phospholipids of phorbol-ester-differentiated U937 cells was investigated. Initially, AA incorporated primarily into choline glycerophospholipids (PC), whereas EPA incorporated mainly into ethanolamine glycerophospholipids (PE). Bromoenol lactone (BEL), an inhibitor of the Group VI Ca2+-independent phospholipase A2 (iPLA2), diminished both lysophosphatidylcholine levels and the incorporation of AA into phospholipids. However BEL had little effect on EPA incorporation. In concanavalin A-activated cells, EPA, but not AA, incorporation was also affected by methyl arachidonyl fluorophosphonate (MAFP), suggesting an additional role for the group IV cytosolic phospholipase A2. In the activated cells AA and EPA did not compete with each other for incorporation, indicating that the pathways for AA and EPA incorporation are partially different. The AA and EPA initially incorporated into PC slowly moved to PE in a process that took several hours. The transfer of AA and EPA from PC to PE was not inhibited by BEL, MAFP or LY311727 [3-(3-acetamide 1-benzyl-2-ethylindolyl-5-oxy)propanesulphonic acid], raising the possibility that an as-yet-undetermined phospholipase A2 may be involved in fatty acid phospholipid remodelling. A strong candidate to be involved in these reactions is a novel Ca2+-independent phospholipase A2 that, unlike all known iPLA2s, is resistant to inhibition by BEL and also to MAFP and LY311727. The enzyme activity cleaves both PC and PE and is thus able to provide the lysoPC and lysoPE acceptors required for the fatty acid acylation reactions.
Abbreviations used: AA, arachidonic acid; Con A, concanavalin A; EPA, eicosa-5,8,11,14,17-pentaenoic acid; PLA2, phospholipase A2; cPLA2, cytosolic phospholipase A2; iPLA2, Ca2+-independent phospholipase A2; sPLA2, secretory phospholipase A2; CoA-IT, CoA-independent transacylase; BEL, bromoenol lactone [(E)-6-(bromomethylene)tetrahydro-3-(1-naphthalenyl)-2H-pyran-2-one]; MAFP, methyl arachidonyl fluorophosphonate; IHP, diethyl 7-(3,4,5-triphenyl-2-oxo-2,3-dihydroimidazol-1-yl)hepatine phosphonate (SK&F 98625); LY311727, 3-(3-acetamide-1-benzyl-2-ethylindolyl-5-oxy)propanesulphonic acid; PC, choline glycerophospholipids; PE, ethanolamine glycerophospholipids; PI, phosphatidylinositol.
This work is dedicated to the memory of Belén Fernández-Boya.