title: Discovery of an arachidonoyl coenzyme A synthetase in human platelets.
abstract: Platelets contain small amounts of a variety of free fatty acids but essentially no free arachidonate. When free fatty acids are incubated with platelets, there is preferential incorporation of arachidonic acid and 8,-11,14-eicosatrienoic acid compared to other fatty acids. We now explain these findings by the discovery that platelets contain two long chain acyl-CoA synthetases. One shows activity with a range of different fatty acids, similar to long chain acyl-CoA synthetases studied previously. A crude platelet membrane preparation contains this enzyme that catalyzes the formation of 0.75 nmol of oleoyl-CoA/min/10(9) platelets. The other enzyme is specific for the prostaglandin precursors arachidonic acid and 8,11,14-eicosatrienoic acid. Based on the ability of fatty acids to inhibit arachidonate and 8,11,14-eicosatrienoate activation, we conclude that other fatty acids including linoleic, 5,8,11-eicosatrienoic, and oleic acids are not substrates for the enzyme. Platelet membranes catalyze formation of 2.9 nmol of arachidonoyl-CoA/min/10(9) platelets and 2.5 nmol of 8,11,14-eicosatrienoyl-CoA/min/10(9) platelets. Arachidonoyl-CoA synthetase has optimal activity at pH 8 and requires ATP (Km = 0.5 mM), Mg2+ (Km = 2.5 mM), CoA (Km = 0.13 mM), and arachidonic acid (Km = 0.03 mM). We propose that the arachidonate-specific acyl-CoA synthetase may control the level of free arachidonic acid in platelets, limiting prostaglandin synthesis by the unstimulated cell and capturing free arachidonate from extracellular sources.
mesh_terms: D000818:Animals; D001792:Blood Platelets / Q000201:enzymology*; D002462:Cell Membrane / Q000201:enzymology; D003066:Coenzyme A Ligases / Q000097:blood* / Q000378:metabolism; D005231:Fatty Acids, Unsaturated / Q000494:pharmacology; D006801:Humans; D007700:Kinetics; D008862:Microsomes, Liver / Q000201:enzymology; D051381:Rats; D013379:Substrate Specificity
chemical_list: D005231:Fatty Acids, Unsaturated; D003066:Coenzyme A Ligases; C033896:arachidonate - CoA ligase