Regulation of eicosanoid synthesis in microvessel endothelium: glucocorticoids do not affect arachidonyl CoA synthetase activity.
The properties of acyl coenzyme A (CoA) synthetase activity were characterized in cultured rabbit coronary microvessel endothelial cells. We report here that microvessel endothelial cells contain two long-chain acyl CoA synthetases. One shows activity with a variety of fatty acids, similar to long-chain non-selective fatty acyl CoA synthetases described previously. The other activity was selective for arachidonic acid and other structurally related substrates. Both activities required ATP, Mg2+ and CoA for optimal activity. The arachidonyl CoA and the non-selective acyl CoA synthetases showed different thermolabilities. Arachidonyl CoA formation was inhibited by greater than 50% after 1 min at 45 degrees C, whereas a 15 min heating treatment was necessary to produce the same relative inhibition of oleoyl CoA synthesis. Glucocorticoid pretreatment (10(-7) M dexamethasone) of the RCME cells did not affect the apparent Km or Vmax, nor the fatty acid selectivity for either acyl CoA synthetase. Therefore, although fatty acyl CoA synthetases may be involved in limiting eicosanoid formation, these activities do not appear to be glucocorticoid-responsive.
Gerritsen
M E
ME
Department of Physiology, New York Medical College, Valhalla 10595.
Perry
C A
CA
D000255:Adenosine Triphosphate / Q000494:pharmacology; D000818:Animals; D001667:Binding, Competitive; D002478:Cells, Cultured; D003065:Coenzyme A / Q000494:pharmacology; D003066:Coenzyme A Ligases / Q000378:metabolism*; D003331:Coronary Vessels; D003907:Dexamethasone / Q000494:pharmacology*; D015777:Eicosanoids / Q000096:biosynthesis*; D004730:Endothelium, Vascular / Q000187:drug effects / Q000378:metabolism*; D005227:Fatty Acids / Q000378:metabolism / Q000494:pharmacology; D006358:Hot Temperature; D008274:Magnesium / Q000494:pharmacology; D019301:Oleic Acid; D009829:Oleic Acids / Q000378:metabolism; D011817:Rabbits; D013379:Substrate Specificity
D015777:Eicosanoids; D005227:Fatty Acids; D009829:Oleic Acids; D019301:Oleic Acid; D003907:Dexamethasone; D000255:Adenosine Triphosphate; D003066:Coenzyme A Ligases; C033896:arachidonate - CoA ligase; D008274:Magnesium; D003065:Coenzyme A