Is platelet-activating factor (PAF-acether) synthesis by murine peritoneal cells (PC) a two-step process?
PAF-acether (1-O-alkyl-2-acetyl-sn-glyceryl-3-phosphorylcholine) is released from several cell sources simultaneously with an inactive non-acylated compound (1-O-alkyl-glyceryl-3-phosphorylcholine) (lyso-PAF-acether). Formation of the latter probably results from the activation of a phospholipase A2 (PLA2). Indeed, the PLA2 inhibitors, bromophenacyl bromide (BPB), mepacrine, 874CB (100 microM), and EDTA (5 mM), blocked the zymosan-induced release of PAF-acether from PC. EDTA and BPB also markedly reduced the release of lyso-PAF-acether. To verify this hypothesis, acetyl coenzyme A (acetyl-CoA) was added to stimulated PC. This enhanced the release of PAF-acether in a dose-dependent fashion from 1 microM acetyl-CoA to reach a maximal increase - 200% - at 100 microM. Furthermore, using 3H acetyl-CoA, incorporation of labelled acetate into PAF-acether was suggested by (1) identical chromatographic patterns of biological activity and radioactivity; (2) disappearance of these activities after treatment with PLA2, but not after exposure to lipase from Rhizopus arrhizus. PAF-acether was also obtained when both acetyl-CoA (100 microM) and synthetic lyso-PAF-acether (0.2 microM) were added to unstimulated PC previously treated with BPB (100 microM for 10 min). These results suggest that the release of PAF-acether is the consequence of at least two different steps: (1) hydrolysis of 1-O-alkyl-2-acyl-glyceryl phosphorylcholine by PLA2; (2) enzymatic acetylation of the hydrolysis product.
D000098:Acetophenones / Q000494:pharmacology; D000105:Acetyl Coenzyme A / Q000378:metabolism; D000123:Acetyltransferases / Q000378:metabolism; D000818:Animals; D001202:Ascitic Fluid / Q000166:cytology; D066298:In Vitro Techniques; D008244:Lysophosphatidylcholines / Q000096:biosynthesis* / Q000378:metabolism; D010741:Phospholipases A / Q000378:metabolism; D054467:Phospholipases A2; D010972:Platelet Activating Factor; D051381:Rats
D000098:Acetophenones; D008244:Lysophosphatidylcholines; D010972:Platelet Activating Factor; C026564:stearoyl alpha-lysolecithin; D000105:Acetyl Coenzyme A; D000123:Acetyltransferases; D010741:Phospholipases A; D054467:Phospholipases A2; C006821:4-bromophenacyl bromide