Studies on NADPH-dependent chloral hydrate reducing enzymes in rat liver cytosol

Mikiko Ikeda, Miho Ezaki, Susumu Kokeguchi, Shinji Ohmori

Research output: Contribution to journalArticle

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Abstract

Chloral hydrate, a sedative hypnotic and also a major metabolite of trichloroethylene in higher animals, is reduced to trichloroethanol by liver extracts. The reducing activity in rat liver cytosol could be separated into four fractions [one NADH- (F1) and three NADPH-dependent (F2, F3 and F4)] by DEAE-cellulose column chromatography. By several procedures, F2 was purified over 1000-fold and F4 was purified over 600-fold from liver cytosol. As judged from polyacrylamide gel electrophoresis performed with and without the addition of sodium dodecylsulfate, the final preparations were essentially homogeneous. They differed in molecular weight, mobility on polyacrylamide gel electrophoresis, pH optimum, substrate specificity, and sensitivity to inhibitors. The molecular weights were estimated to be 36,000 and 32,500 for F2 and F4, respectively, by polyacrylamide gel electrophoresis in the presence of sodium dodecylsulfate. The estimation of molecular weights by thin-layer gel chromatography indicated that the enzymes were monomers. An examination of over thirty substrates revealed that both enzymes catalyzed the reduction of long-chain aliphatic, alicyclic and aromatic aldehydes as well as halogenated acetaldehyde. The F2 enzyme acted on d-glucuronate, indicating that it was identical to the aldehyde reductase recently reported by other workers (l-gulonate: NADP+ 1-oxidoreductase EC 1.1.1.19). The F4 enzyme, on the other hand, preferentially acted on C24 3-ketosteroids.

Original languageEnglish
Pages (from-to)1931-1939
Number of pages9
JournalBiochemical Pharmacology
Volume30
Issue number14
DOIs
Publication statusPublished - Jul 15 1981

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Chloral Hydrate
NADP
Liver
Cytosol
Rats
Electrophoresis
Polyacrylamide Gel Electrophoresis
Molecular Weight
Molecular weight
L-glucuronate reductase
Enzymes
Sodium
Ketosteroids
Trichloroethylene
Liver Extracts
DEAE-Cellulose Chromatography
Aldehyde Reductase
DEAE-Cellulose
Glucuronic Acid
Column chromatography

ASJC Scopus subject areas

  • Pharmacology

Cite this

Studies on NADPH-dependent chloral hydrate reducing enzymes in rat liver cytosol. / Ikeda, Mikiko; Ezaki, Miho; Kokeguchi, Susumu; Ohmori, Shinji.

In: Biochemical Pharmacology, Vol. 30, No. 14, 15.07.1981, p. 1931-1939.

Research output: Contribution to journalArticle

Ikeda, Mikiko ; Ezaki, Miho ; Kokeguchi, Susumu ; Ohmori, Shinji. / Studies on NADPH-dependent chloral hydrate reducing enzymes in rat liver cytosol. In: Biochemical Pharmacology. 1981 ; Vol. 30, No. 14. pp. 1931-1939.
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