Biosynthesis of prostaglandins in rabbit kidney medulla. Properties of prostaglandin synthase.
A simple radioactive-substrate assay for prostaglandin synthase (EC 1.14.99.1), which uses t.l.c. to measure simultaneously different prostaglandins synthesized from one precursor substrate, was developed. Rabbit kidney-medulla prostaglandin synthase catalyses the formation of prostaglandin E2, prostaglandin F2alpha and prostaglandin D2 from arachidonic acid. Fractionation of crude homogenates indicated that the microsomal fraction possessed the highest specific activity of prostaglandin synthase, whereas the soluble fraction exhibited little enzyme activity but rather contained a heat-labile inhibitory macromolecular factor(s), which might be attributed to the serum albumin present in this fraction. The microsomal fraction possessed low intrinsic enzyme activity, but the actvity could be fully stimulated by the presence of both GSH (reduced glutathione) and a phenolic cofactor. Only cysteine could partially replace GSH, whereas other thiols were inactive and some were even inhibitory. A variety of phenolic compounds, including catecholamines, dopamine (3,4-dihydroxyphenethylamine), 5-hydroxytryptamine and quinol, were active in stimulating prostaglandin synthase. In all cases, the stimulation was reflected in the synthesis of all three prostaglandins with ratios not significantly altered by different phenolic cofactors. The synthesis of each of the different prostaglandins appeared to have similar pH optima. The enzyme system was not inhibited by thiol-group inhibitors or a variety of metal chelators except for cyanide and 8-hydroxyquinoline. Characterization of the kidney-medulla prostaglandin synthase system indicated that it exhibited properties similar to those of the enzyme system present in seminal vesicles.
D000818:Animals; D001095:Arachidonic Acids / Q000378:metabolism; D002614:Chelating Agents / Q000494:pharmacology; D016861:Cyclooxygenase Inhibitors; D004837:Epinephrine / Q000494:pharmacology; D005978:Glutathione / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D007668:Kidney / Q000378:metabolism*; D007679:Kidney Medulla / Q000201:enzymology / Q000378:metabolism*; D007700:Kinetics; D008861:Microsomes / Q000201:enzymology; D006899:Mixed Function Oxygenases / Q000378:metabolism*; D010636:Phenols / Q000494:pharmacology; D011451:Prostaglandin-Endoperoxide Synthases / Q000378:metabolism*; D011453:Prostaglandins / Q000096:biosynthesis*; D011817:Rabbits; D012709:Serum Albumin / Q000494:pharmacology; D013438:Sulfhydryl Compounds / Q000494:pharmacology
D001095:Arachidonic Acids; D002614:Chelating Agents; D016861:Cyclooxygenase Inhibitors; D010636:Phenols; D011453:Prostaglandins; D012709:Serum Albumin; D013438:Sulfhydryl Compounds; D006899:Mixed Function Oxygenases; D011451:Prostaglandin-Endoperoxide Synthases; D005978:Glutathione; D004837:Epinephrine