Acyl-chain selectivity of the 85 kDa phospholipase A2 and of the release process in intact macrophages.
The selectivity of the intracellular 85 kDa phospholipase A2 (PLA2-85) towards fatty acids closely related to arachidonic acid has been investigated, using purified PLA2-85 from J774 cells and mixed phospholipids, dually acyl-chain-labelled in the sn-2 position. In parallel experiments, we assessed the acyl-chain selectivity of the release process in intact, dually labelled, peritoneal mouse macrophages responding to either calcium ionophore or zymosan beads in the presence of indomethacin and BSA. The results obtained in the two systems were very similar, which supports previous evidence that PLA2-85 is responsible for stimulus-induced release of eicosanoid precursor in mouse macrophages. In the in vitro system, PLA2-85 was found to exhibit a moderate selectivity towards C20 acyl chains differing in double-bond structure, while the sensitivity to acyl-chain length was more pronounced. Together with previous data, these results demonstrate a striking preference for C20 over either C18 or C22 unsaturated acyl chains.
Sundler
R
R
Department of Medical and Physiological Chemistry, Lund University, Sweden.
Winstedt
D
D
Wijkander
J
J
D000818:Animals; D016718:Arachidonic Acid / Q000378:metabolism; D000001:Calcimycin / Q000494:pharmacology; D002460:Cell Line; D002478:Cells, Cultured; D005227:Fatty Acids / Q000378:metabolism*; D007213:Indomethacin / Q000494:pharmacology; D008081:Liposomes / Q000378:metabolism; D017737:Macrophages, Peritoneal / Q000187:drug effects / Q000201:enzymology*; D051379:Mice; D008970:Molecular Weight; D010714:Phosphatidylethanolamines / Q000378:metabolism; D010716:Phosphatidylinositols / Q000378:metabolism; D010741:Phospholipases A / Q000378:metabolism*; D054467:Phospholipases A2; D010743:Phospholipids / Q000378:metabolism; D012710:Serum Albumin, Bovine / Q000494:pharmacology; D013379:Substrate Specificity; D015054:Zymosan / Q000494:pharmacology
D005227:Fatty Acids; D008081:Liposomes; D010714:Phosphatidylethanolamines; D010716:Phosphatidylinositols; D010743:Phospholipids; D012710:Serum Albumin, Bovine; D016718:Arachidonic Acid; D000001:Calcimycin; D015054:Zymosan; D010741:Phospholipases A; D054467:Phospholipases A2; D007213:Indomethacin