Synthesis of [5,6-3H]arachidonic acid and its use in development of a sensitive assay for prostacyclin synthetase.
[5,6-3H]arachidonic acid has been prepared by catalytic reduction of eicosa-cis-8,11,14-trien-5-ynoic acid (IV) over Lindlar catalyst. When either [5,6-3H]arachidonic acid or [5,6-3H]PGH2 is converted into PGI2 by swine aortic microsomes, the tritium at C-6 is lost to the medium. Thus, the progress of this enzymic rearrangement may be monitored by following the rate of tritium release. As swine aortic microsomes contain only low levels of cyclooxygenase, it is necessary to fortify the system with ram seminal vesicular microsomes (rich in cyclooxygenase) when [5,6-3H]arachidonic acid is used as the indirect substrate.
D015126:8,11,14-Eicosatrienoic Acid / Q000031:analogs & derivatives; D000818:Animals; D001011:Aorta / Q000201:enzymology; D001095:Arachidonic Acids / Q000138:chemical synthesis*; D003577:Cytochrome P-450 Enzyme System; D011464:Epoprostenol / Q000032:analysis / Q000096:biosynthesis*; D019746:Intramolecular Oxidoreductases; D007553:Isotope Labeling / Q000379:methods; D007700:Kinetics; D009682:Magnetic Resonance Spectroscopy; D008861:Microsomes / Q000201:enzymology; D011453:Prostaglandins / Q000096:biosynthesis*; D014316:Tritium
D001095:Arachidonic Acids; D011453:Prostaglandins; D014316:Tritium; C025421:eicosa-8,11,14-trien-5-ynoic acid; D003577:Cytochrome P-450 Enzyme System; D011464:Epoprostenol; D019746:Intramolecular Oxidoreductases; C022461:prostacyclin synthetase; D015126:8,11,14-Eicosatrienoic Acid