title: Substrate specificities of neuronal nuclear acetyltransferases involved in the synthesis of platelet-activating factor: differences in the use of 1-alkyl and 1-acyl lysophospholipid acceptors.
abstract: The selectivity of alkylglycerophosphate (AGP) acetyltransferase and lyso-platelet-activating factor (lyso-PAF) acetyltransferase was studied in neuronal nuclei isolated from cerebral cortices of 15-day-old rabbits. Specifically, 1-alkyl and 1-acyl analogues were compared as acceptors in these acetylation reactions. A number of observations supported one nuclear activity in the acetylation of AGP and lyso-PA. Lyso-PA was a competitive substrate for AGP, Km values for AGP and lyso-PA were similar, as were acetylation rates measured at individual AGP or lyso-PA concentrations, and the acetylation of both substrates was unaffected by preincubations with protein phosphatase 1 (PP-1). In contrast, there were a number of differences seen in the acetylation of lyso-PAF and lyso-PC. The kinetics for lyso-PC acetylation (as a function of lyso-PC concentration) were not hyperbolic, and lyso-PC was not a competitive substrate for the acetylation of lyso-PAF. Unlike acetylation rates with lyso-PAF, lyso-PC acetylation was not reduced by preincubations with PP-1, and was less susceptible to inhibition particularly at high levels of free fatty acid. In addition, rates of acetylation of lyso-PC were selectively increased by the presence of lyso-PA. When neuronal nuclear envelope fractions (NE) were prepared from N1, the specific acetylation activity with lyso-PAF was significantly lower in NE, while the activities for lyso-PC were comparable in NE and the parent N1 fraction. The results with the acetylation of lyso-PC and lyso-PAF suggest that the lyso-PC acetyltransferase may be in a uniquely sequestered state within the neuronal nucleus. This could explain the smaller inhibition of lyso-PC acetylation by free fatty acid, the maintenance of lyso-PC acetylation during PP-1 preincubations, the non-hyperbolic response to lyso-PC concentrations and the selective preservation of lyso-PC acetylation during NE isolation. This protected status could result from a more internal location for this acetyltransferase within the membranes of the nuclear envelope, or possibly an association of the enzyme with the nuclear matrix that is disrupted with the exposure of N1 to lyso-PA.
authors.affiliation: Division of Neurology, Department of Medicine, Clinical Science Division, Room 6368, Medical Sciences Bldg., University of Toronto, Toronto, Ont., Canada.
authors.forename: R
authors.initials: R
authors.lastname: Roy Baker
authors.forename: H
authors.initials: H
authors.lastname: Chang
mesh_terms: D000107:Acetylation; D000123:Acetyltransferases / Q000378:metabolism*; D000214:Acyl Coenzyme A / Q000494:pharmacology; D000818:Animals; D001921:Brain / Q000201:enzymology*; D002467:Cell Nucleus / Q000201:enzymology; D005994:Glycerophosphates / Q000378:metabolism; D007700:Kinetics; D008244:Lysophosphatidylcholines / Q000378:metabolism; D008246:Lysophospholipids / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D009474:Neurons / Q000201:enzymology; D009685:Nuclear Envelope / Q000378:metabolism; D009687:Nuclear Proteins / Q000378:metabolism; D019301:Oleic Acid / Q000494:pharmacology; D010972:Platelet Activating Factor / Q000031:analogs & derivatives / Q000096:biosynthesis* / Q000378:metabolism; D011817:Rabbits; D013379:Substrate Specificity
chemical_list: D000214:Acyl Coenzyme A; D005994:Glycerophosphates; D008244:Lysophosphatidylcholines; D008246:Lysophospholipids; D009687:Nuclear Proteins; C029271:O-deacetyl platelet activating factor; D010972:Platelet Activating Factor; C017585:oleoyl-coenzyme A; D019301:Oleic Acid; D000123:Acetyltransferases; C052591:alkylglycerophosphate 2-O-acetyltransferase