Preparation of oxygen-18-labeled lipoxygenase metabolites of arachidonic acid.
Plasma pseudocholinesterase and porcine liver esterase were used to catalyse the incorporation of the stable isotope oxygen-18 into the carboxyl moiety of lipoxygenase metabolites of arachidonic acid. This simple method produces eicosanoid products containing two oxygen-18 atoms; but the enzymes studied were found to display large substrate specificity in the efficiencies at which oxygen-18 could be incorporated into the lipoxygenase metabolites. Furthermore, [18O2]LTB4 was found not to back exchange during in vitro incubation with human neutrophils. The methods involved for stable isotope incorporation are simple, efficient and produce highly enriched species in a short time. By varying the type of esterase, the amount of esterase or the length of incubation highly enriched species of all eicosanoids tested could be prepared.
D000818:Animals; D016718:Arachidonic Acid; D001095:Arachidonic Acids / Q000378:metabolism*; D004950:Esterases / Q000378:metabolism; D005227:Fatty Acids / Q000378:metabolism*; D008401:Gas Chromatography-Mass Spectrometry; D006893:Hydroxyeicosatetraenoic Acids / Q000378:metabolism; D066298:In Vitro Techniques; D007975:Leukotriene B4 / Q000378:metabolism; D008084:Lipoxygenase / Q000378:metabolism*; D009504:Neutrophils / Q000378:metabolism; D010104:Oxygen Radioisotopes; D011453:Prostaglandins / Q000378:metabolism; D013552:Swine
D001095:Arachidonic Acids; D005227:Fatty Acids; D006893:Hydroxyeicosatetraenoic Acids; D010104:Oxygen Radioisotopes; D011453:Prostaglandins; D007975:Leukotriene B4; D016718:Arachidonic Acid; D008084:Lipoxygenase; D004950:Esterases