On the relation between a stearoyl-specific transacylase from bovine testis membranes and a copurifying acyltransferase.
Bovine testis membranes contain a coenzyme A-dependent transacylase that can catalyze the preferential transfer of stearoyl groups from phosphoglycerides to sn-2-acyl molecular species of lysophosphatidic acid and lysophosphatidylinositol [Itabe et al., (1992) J. Biol. Chem. 267, 15319-15325]. We have now purified this enzyme 1000-fold and shown that it copurifies with an acyltransferase. The purified transacylase can use phosphatidic acid, phosphatidylinositol, or phosphatidylinositol-4-phosphate as an acyl donor and catalyzes the transfer of stearoyl groups in preference to palmitoyl groups or oleoyl groups. In contrast, the purified acyltransferase uses acyl-coenzyme A as an acyl donor and shows no such preference for stearoyl group transfer. Furthermore, phosphatidylinositol-4, 5-bisphosphate inhibits the two enzymes to different extents and by different mechanisms. Nevertheless, the enzymes are similar in several respects: they use the same acyl acceptors and, when assayed together, compete for the acyl acceptor, sn-2-oleoyl lysophosphatidic acid; they lose activity in parallel unless stabilized by the addition of an anionic phosphoglyceride or stearoyl-coenzyme A; and they show similar sensitivities to heat and pH. One way to explain these results is to postulate that the transacylase reaction occurs in two successive steps: a stearoyl-specific first step in which a stearoyl group is transferred from an sn-1-stearoyl-2-acyl phosphoglyceride to coenzyme A, and a relatively non-acyl-chain-specific second step in which a stearoyl group is transferred from stearoyl-coenzyme A to an sn-2-acyl lysophosphoglyceride. According to this line of reasoning, the transacylase assay that we have used measures the net effect of both steps, whereas the acyltransferase assay measures only the effect of the second step.
Hollenback
D
D
Howard Hughes Medical Institute, Department of Medicine, University of Washington, Seattle 98195-7370, USA.
Glomset
J A
JA
D000217:Acyltransferases / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000378:metabolism*; D000818:Animals; D001667:Binding, Competitive; D002417:Cattle; D002462:Cell Membrane / Q000201:enzymology; D004789:Enzyme Activation; D004795:Enzyme Stability / Q000187:drug effects; D008297:Male; D010712:Phosphatidic Acids / Q000378:metabolism; D019269:Phosphatidylinositol 4,5-Diphosphate / Q000494:pharmacology; D018129:Phosphatidylinositol Phosphates / Q000378:metabolism; D010716:Phosphatidylinositols / Q000378:metabolism; D013379:Substrate Specificity; D013737:Testis / Q000201:enzymology*
D010712:Phosphatidic Acids; D019269:Phosphatidylinositol 4,5-Diphosphate; D018129:Phosphatidylinositol Phosphates; D010716:Phosphatidylinositols; C037178:phosphatidylinositol 4-phosphate; D000217:Acyltransferases; C075831:stearoyl transacylase