TY - JOUR
T1 - Formation of 20-oxoleukotriene B4 by an alcohol dehydrogenase isolated from human neutrophils
AU - Gotoh, Yoichi
AU - Sumimoto, Hideki
AU - Minakami, Shigeki
N1 - Funding Information:
We wish to thank Dr. Ryuichi Isobe (Center of Advanced Instrumental Analysis, Faculty of Pharmaceutical Science, Kyushu University) for GC-MS analysis. This work was supported in part by grants from the Ministry of Education, Science and Culture, Japan.
PY - 1990/3/12
Y1 - 1990/3/12
N2 - 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 μM and the Vmax is 2.04 μmol/min per mg of protein. The activity is inhibited by ω-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.
AB - 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 μM and the Vmax is 2.04 μmol/min per mg of protein. The activity is inhibited by ω-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.
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U2 - 10.1016/0005-2760(90)90109-B
DO - 10.1016/0005-2760(90)90109-B
M3 - Article
C2 - 2155662
AN - SCOPUS:0025058882
SN - 0005-2760
VL - 1043
SP - 52
EP - 56
JO - Biochimica et Biophysica Acta (BBA)/Lipids and Lipid Metabolism
JF - Biochimica et Biophysica Acta (BBA)/Lipids and Lipid Metabolism
IS - 1
ER -