Formation of 20-oxoleukotriene B4 by an alcohol dehydrogenase isolated from human neutrophils

Yoichi Gotoh, Hideki Sumimoto, Shigeki Minakami

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)52-56
Number of pages5
JournalBiochimica et Biophysica Acta (BBA)/Lipids and Lipid Metabolism
Volume1043
Issue number1
DOIs
Publication statusPublished - Mar 12 1990

Fingerprint

Leukotriene B4
Alcohol Dehydrogenase
Neutrophils
NAD
High Pressure Liquid Chromatography
NADH Dehydrogenase
Hydroxy Acids
High performance liquid chromatography
Reverse-Phase Chromatography
20-aldehyde leukotriene B4
NADP
Gas chromatography
Gas Chromatography-Mass Spectrometry
Spectrometry
Mass spectrometry
Spectrum Analysis
Oxidoreductases
Fatty Acids
Acids
hydroxide ion

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Biochemistry
  • Endocrinology

Cite this

Formation of 20-oxoleukotriene B4 by an alcohol dehydrogenase isolated from human neutrophils. / Gotoh, Yoichi; Sumimoto, Hideki; Minakami, Shigeki.

In: Biochimica et Biophysica Acta (BBA)/Lipids and Lipid Metabolism, Vol. 1043, No. 1, 12.03.1990, p. 52-56.

Research output: Contribution to journalArticle

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