leukotriene-b4 and 12-hydroxydodecanoic-acid

When 20-hydroxyleukotriene B4 (20-OH-LTB4) is incubated at pH 10.5 in the presence of NAD+ with an alcohol dehydrogenase isolated from human neutrophils, a polar product is formed as detected on reverse-phase high-performance liquid chromatography (RP-HPLC). The product is identified as 20-oxo-LTB4 (20-CHO-LTB4) on the basis of its co-elution with the authentic compound on HPLC, ultraviolet spectrometry and gas chromatography-mass spectrometry. The 20-CHO-LTB4-forming activity requires NAD+, but NADP+ scarcely replaces NAD+. The apparent Km for 20-OH-LTB4 is 83 microM and the Vmax is 2.04 mumol/min per mg of protein. The activity is inhibited by omega-hydroxy fatty acids such as 12-hydroxylauric acid, 16-hydroxypalmitic acid and 12(S), 20-dihydroxyeicosatetraenoic acid, but not by 4-methylpyrazole. At pH 7.0 with NADH, the purified dehydrogenase catalyzes the reverse reaction, the reduction of 20-CHO-LTB4 to 20-OH-LTB4.

Topics: 12-Hydroxy-5,8,10,14-eicosatetraenoic Acid; Alcohol Dehydrogenase; Chromatography, High Pressure Liquid; Gas Chromatography-Mass Spectrometry; Humans; Hydrogen-Ion Concentration; Hydroxyeicosatetraenoic Acids; Kinetics; Lauric Acids; Leukotriene B4; NAD; NADP; Neutrophils; Palmitic Acids; Spectrophotometry, Ultraviolet