Formation of 20-oxoleukotriene B4 by an alcohol dehydrogenase isolated from human neutrophils.
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.
Gotoh
Y
Y
Department of Biochemistry, Kyushu University School of Medicine, Fakuoka, Japan.
Sumimoto
H
H
Minakami
S
S
D019377:12-Hydroxy-5,8,10,14-eicosatetraenoic Acid / Q000031:analogs & derivatives*; D000426:Alcohol Dehydrogenase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D008401:Gas Chromatography-Mass Spectrometry; D006801:Humans; D006863:Hydrogen-Ion Concentration; D006893:Hydroxyeicosatetraenoic Acids / Q000494:pharmacology; D007700:Kinetics; D007850:Lauric Acids / Q000494:pharmacology; D007975:Leukotriene B4 / Q000378:metabolism* / Q000031:analogs & derivatives; D009243:NAD / Q000494:pharmacology; D009249:NADP / Q000494:pharmacology; D009504:Neutrophils / Q000201:enzymology*; D010169:Palmitic Acids / Q000494:pharmacology; D013056:Spectrophotometry, Ultraviolet
C040735:12,20-dihydroxy-5,8,10,14-eicosatetraenoic acid; D006893:Hydroxyeicosatetraenoic Acids; D007850:Lauric Acids; D010169:Palmitic Acids; D009243:NAD; C056168:20-aldehyde leukotriene B4; D007975:Leukotriene B4; C031263:12-hydroxydodecanoic acid; D009249:NADP; D019377:12-Hydroxy-5,8,10,14-eicosatetraenoic Acid; C063407:16-hydroxypalmitic acid; D000426:Alcohol Dehydrogenase