title: A novel CoA-independent transacetylase produces the ethanolamine plasmalogen and acyl analogs of platelet-activating factor (PAF) with PAF as the acetate donor in HL-60 cells.
abstract: In this study, we demonstrate the presence of a unique membrane-associated transacetylase that transfers the acetate group from platelet-activating factor (PAF) to lysoplasmalogen (in the presence of EDTA and sodium acetate) with the formation of 1-alk-1-enyl-2-acetyl-sn-glycero-3-phosphoethanolamine (alk-1-enylacetyl-GPE). The identity of alk-1-enylacetyl-GPE was confirmed by acid hydrolysis, phospholipases A2 or C treatment and derivatization by fluorodinitrobenzene. The transacetylase has no requirement for Ca2+, Mg2+, or CoA and a broad pH optimum (7.0-8.0) with Km values of 12.0 microM for PAF and 106.4 microM for lysoplasmalogens. The enzyme activity from the isolated membrane fraction is not changed when whole cells are supplemented with 20:4, induced to differentiate into granulocytes, or treated with ionophore A23187. Radyllyso-sn-glycero-3-phosphocholine (GPC), radyllyso-GPE, acyllyso-sn-glycero-3-phosphoserine (GPS), acyllyso-sn-glycero-3-phosphoinositol (GPI), alkyllyso-sn-glycero-3-phosphate (GP), acyllyso-GP, or cis-9-octadecen-1-ol can also serve as acetate acceptors, whereas alkylglycerol, acylglycerol, or cholesterol are inactive. Differences in substrate acceptor specificity, sensitivity toward phenylmethylsulfonyl fluoride, and response to temperature suggest that the CoA-independent transacetylase and the CoA-independent transacylase that transfers long-chain acyl moieties are two separate enzymes. With intact differentiated HL-60 cells, [3H]acetate from [3H]PAF can be incorporated into alk-1-enylacetyl-GPE in the presence of ionophore A23187, but not in its absence. Moreover, phospholipase A2 inhibitors (p-bromophenacyl bromide and mepacrine) block the transacetylation process in whole cell system. These results indicate the production of alk-1-enyllyso-GPE is a rate-limiting factor for the subsequent transacetylation step during cell activation. We conclude that the transacetylase may participate in the biosynthesis of ethanolamine plasmalogen and acyl analogs of PAF, in vivo, fine-tuning of PAF biological responses, and cross-talk between de novo and remodeling pathways of PAF biosynthesis.
authors.affiliation: Medical Sciences Division, Oak Ridge Associated Universities, Tennessee 37831-0117.
authors.forename: T C
authors.initials: TC
authors.lastname: Lee
authors.forename: Y
authors.initials: Y
authors.lastname: Uemura
authors.forename: F
authors.initials: F
authors.lastname: Snyder
mesh_terms: D000085:Acetates / Q000378:metabolism*; D000123:Acetyltransferases / Q000378:metabolism*; D016718:Arachidonic Acid / Q000494:pharmacology; D000001:Calcimycin / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism; D002413:Cations, Divalent; D002462:Cell Membrane / Q000201:enzymology; D002478:Cells, Cultured; D002855:Chromatography, Thin Layer; D006863:Hydrogen-Ion Concentration; D008274:Magnesium / Q000378:metabolism; D010664:Phenylmethylsulfonyl Fluoride / Q000494:pharmacology; D010743:Phospholipids / Q000378:metabolism; D010955:Plasmalogens / Q000378:metabolism*; D010972:Platelet Activating Factor / Q000031:analogs & derivatives / Q000378:metabolism*; D013379:Substrate Specificity; D013696:Temperature
chemical_list: D000085:Acetates; D002413:Cations, Divalent; D010743:Phospholipids; D010955:Plasmalogens; D010972:Platelet Activating Factor; C020791:phosphatidal ethanolamines; D016718:Arachidonic Acid; D000001:Calcimycin; D010664:Phenylmethylsulfonyl Fluoride; D000123:Acetyltransferases; D008274:Magnesium; D002118:Calcium