LIVER MITOCHONDRIAL CYTOCHROME-P450 CYP27 AND RECOMBINANT-EXPRESSED HUMAN CYP27 CATALYZE 1-ALPHA-HYDROXYLATION OF 25-HYDROXYVITAMIN D-3

被引：68
作者：
AXEN, E
POSTLIND, H
SJOBERG, H
WIKVALL, K
机构：
[1] Division of Biochemistry, Dept. of Pharmaceutical Biosciences, University of Uppsala
来源：
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA | 1994年 / 91卷 / 21期
关键词：
STEROL; 27-HYDROXYLASE; HEPATIC 1-ALPHA HYDROXYLATION; BACTERIAL EXPRESSION;
D O I：
10.1073/pnas.91.21.10014
中图分类号：
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号：
07 ; 0710 ; 09 ;
摘要：
A cytochrome P450 catalyzing 1 alpha-hydroxylation of 25-hydroxyvitamin D-3 was purified from pig liver mitochondria. It also catalyzed 27-hydroxylation of 25-hydroxyvitamin D-3 and 25-hydroxylation of vitamin D-3 The ratio between the 1 alpha-, 27-, and 25-hydroxylase activities remained essentially constant during the purification. Substrates for sterol 27-hydroxylase CYP27 inhibited and a monoclonal antibody raised against CYP27 immunoprecipitated the 1 alpha-, 27-, and 25-hydroxylase activities. Apparently homogeneous preparations of CYP27 from pig and rabbit liver mitochondria catalyzed 1 alpha-hydroxylation. Human liver mitochondrial CYP27 was expressed from its cDNA in Escherichia coli. The nucleotide sequence encoding the N terminus of CYP27 was modified in the first eight codons to achieve expression in E. coli. The purified recombinant-expressed CYP27 reconstituted with the electron-transferring system of adrenal mitochondria catalyzed 1 alpha-hydroxylation of 25-hydroxyvitamin DJ Expression of unmodified CYP27 cDNA in simian COS cells confirmed the 1 alpha-hydroxylase activity toward 25-hydroxyvitamin D-3.
引用
收藏
页码：10014 / 10018
页数：5
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