Solubilization and partial purification of prostaglandin endoperoxide synthetase of rabbit kidney medulla.
The microsomes of rabbit kidney medulla converted arachidonic acid into prostaglandin E2 in the presence of hemoglobin, tryptophan and glutathione as activators. When themicrosomal suspension was treated with 1% Tween 20, a solubilized enzyme was obtained which catalyzed the conversion of arachidonic acid to prostaglandins G2 and H2. The solubilized enzyme was adsorbed to and then eluted from an omega-aminooctyl Sepharose 4B column, resulting in about 10-fold purification over the microsomes. The partially purified enzyme produced predominantly prostaglandin G2 in the presence of hemoglobin, while prostaglandin H2 was produced in the presence of both hemoglobin and tryptophan. The stimulation of prostaglandin endoperoxide formation was also observed with other heme and aromatic compounds. Prostaglandin H2 synthesis was inhibited by a variety of compounds including non-steroidal anti-inflammatory drugs, thiol compounds and prostaglandin analogues with a thiol group(s).
D000818:Animals; D001095:Arachidonic Acids / Q000378:metabolism; D001241:Aspirin / Q000494:pharmacology; D006454:Hemoglobins / Q000378:metabolism; D007213:Indomethacin / Q000494:pharmacology; D007535:Isomerases / Q000302:isolation & purification / Q000378:metabolism; D007668:Kidney / Q000201:enzymology*; D007679:Kidney Medulla / Q000201:enzymology*; D008861:Microsomes / Q000201:enzymology; D011451:Prostaglandin-Endoperoxide Synthases / Q000302:isolation & purification / Q000378:metabolism*; D011458:Prostaglandins E / Q000096:biosynthesis; D011463:Prostaglandins H / Q000096:biosynthesis; D011817:Rabbits; D012701:Serotonin / Q000494:pharmacology; D012995:Solubility; D013439:Sulfhydryl Reagents / Q000494:pharmacology; D014364:Tryptophan / Q000378:metabolism
D001095:Arachidonic Acids; D006454:Hemoglobins; D011458:Prostaglandins E; D011463:Prostaglandins H; D013439:Sulfhydryl Reagents; D012701:Serotonin; D014364:Tryptophan; D011451:Prostaglandin-Endoperoxide Synthases; D007535:Isomerases; D001241:Aspirin; D007213:Indomethacin