Lipoxygenase-like enzyme in rat testis microsomes.
Microsomes, separated from rat testes, were found capable of oxidizing linoleate and arachidonate. The enzyme activity was solubilized with 1% Triton X-100 in acetate buffer (pH 5.0) and purified by affinity chromatography. The overall purification from the starting preparation was approx. 40-fold. The affinity-purified enzyme was almost homogeneous as determined by electrophoresis in polyacrylamide gel. The enzyme was characterized as lipoxygenase-like from its spectrum, specificity, effect of linoleate on its fluorescence and linoleate oxidation products. Three types of compounds separated by thin-layer chromatography were generally present in the lipoxygenase-like enzyme reaction on linoleic acid: substrate fatty acid, polar by-products and hydroperoxides. The hydroperoxides were analyzed by infrared spectra and mass spectrometry and showed the presence of both 9- and 13-hydroxy isomers.
D000818:Animals; D002846:Chromatography, Affinity; D006863:Hydrogen-Ion Concentration; D008084:Lipoxygenase / Q000302:isolation & purification / Q000378:metabolism*; D016859:Lipoxygenase Inhibitors; D008297:Male; D008861:Microsomes / Q000201:enzymology; D051381:Rats; D013050:Spectrometry, Fluorescence; D013379:Substrate Specificity; D013737:Testis / Q000201:enzymology*
D016859:Lipoxygenase Inhibitors; D008084:Lipoxygenase