Enzyme activity assay for cholesterol 27-hydroxylase in mitochondria
نویسندگان
چکیده
منابع مشابه
Enzyme activity assay for cholesterol 27-hydroxylase in mitochondria.
Mitochondrial cholesterol 27-hydroxylase (CYP27A1) plays an important role in the maintenance of intracellular cholesterol homeostasis. Cholesterol delivery to the mitochondrial inner membrane is believed to be a rate-limiting step for the "acidic" pathway of bile acid synthesis. This work reports that proteinase K treatment of mitochondria markedly increases CYP27A1 specific activity. With end...
متن کاملSynthesis of 27-hydroxycholesterol in rat liver mitochondria: HPLC assay and marked activation by exogenous cholesterol.
Sterol 27-hydroxylase, the mitochondrial enzyme that catalyzes the first step in oxidation of the sterol side chain in hepatic bile acid synthesis, also catalyzes the synthesis of 27-hydroxycholesterol from cholesterol. We have developed a high performance liquid chromatography (HPLC) assay for this enzyme, using either endogenous or exogenous cholesterol as substrate and cholesterol oxidase to...
متن کاملAtherosclerosis and sterol 27-hydroxylase: evidence for a role of this enzyme in elimination of cholesterol from human macrophages.
27-Hydroxycholesterol was found in surprisingly high amounts in atherosclerotic human femoral arteries. When human macrophages were cultured in a medium containing serum, there was a significant transfer of 27-hydroxy-cholesterol and 3 beta-hydroxy-5-cholestenoic acid from the cells into the medium. Sterol 27-hydroxylase (EC 1.14.13.15) is likely to be responsible for formation of the two produ...
متن کاملCholesterol 26-hydroxylase activity of hamster liver mitochondria: isotope ratio analysis using deuterated 26-hydroxycholesterol.
Deuterated 26-hydroxycholesterol prepared from diosgenin by modifications of existing methods permitted the determination of mitochondrial cholesterol 26-hydroxylase using endogenous cholesterol as the substrate. Enzyme activity in a group of Syrian hamsters was found to be 10.3 +/- 3.7 pmol.min-1.mg protein-1.
متن کاملEnzymatic assay for cholesterol ester hydrolase activity.
A rapid and accurate method is described for the assay of cholesterol ester hydrolase (CEH) activity. Aliquots of the enzyme-substrate incubation mixture are extracted into isopropanol. The free cholesterol concentration in each extract is determined enzymatically using a single aqueous reagent containing cholesterol oxidase and peroxidase. The free cholesterol remaining after the cholesterol e...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
ژورنال
عنوان ژورنال: Journal of Lipid Research
سال: 2006
ISSN: 0022-2275
DOI: 10.1194/jlr.m600117-jlr200