An improved method for assay of cholesterol 7α-hydroxylase activity

Tadashi Ogishima, Kyuichiro Okuda

Research output: Contribution to journalArticle

89 Citations (Scopus)

Abstract

An improved assay method for cholesterol 7α-hydroxylase which is accurate, sensitive and yet still simple is described. The method consists of two parts: the first part is cholesterol 7α-hydroxylation in liver microsomes utilizing cholesterol in situ as the substrate, and the second part is conversion of the product, 7α-hydroxycholesterol, into 7α-hydroxy-4-cholesten-3-one having an intense absorption at 240 nm by the action of cholesterol oxidase. The converted sterol is then analyzed by high-performance liquid chromatography. During the second enzyme reaction, the first enzyme reaction is halted and further metabolism of the product is prevented. In consequence, the method had more than 10-fold increase in the sensitivity than the previous one.

Original languageEnglish
Pages (from-to)228-232
Number of pages5
JournalAnalytical Biochemistry
Volume158
Issue number1
DOIs
Publication statusPublished - Jan 1 1986
Externally publishedYes

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Cholesterol 7-alpha-Hydroxylase
Assays
Cholesterol Oxidase
Cholesterol
Hydroxycholesterols
Hydroxylation
High performance liquid chromatography
Sterols
Enzymes
Metabolism
Liver
Liver Microsomes
Substrates
High Pressure Liquid Chromatography
7 alpha-hydroxy-4-cholesten-3-one

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

An improved method for assay of cholesterol 7α-hydroxylase activity. / Ogishima, Tadashi; Okuda, Kyuichiro.

In: Analytical Biochemistry, Vol. 158, No. 1, 01.01.1986, p. 228-232.

Research output: Contribution to journalArticle

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