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编码大鼠11β-羟化酶[细胞色素P450(11β)]和醛固酮合酶[细胞色素P450(11β,aldo)]的cDNA的功能表达。

Functional expression of the cDNAs encoding rat 11 beta-hydroxylase [cytochrome P450(11 beta)] and aldosterone synthase [cytochrome P450(11 beta, aldo)].

作者信息

Nonaka Y, Okamoto M

机构信息

Department of Biochemistry, Osaka University Medical School, Japan.

出版信息

Eur J Biochem. 1991 Dec 18;202(3):897-902. doi: 10.1111/j.1432-1033.1991.tb16449.x.

DOI:10.1111/j.1432-1033.1991.tb16449.x
PMID:1765101
Abstract

Expression plasmids containing two cDNAs of a rat cytochrome P450(11 beta) family, pcP450(11 beta)-62 [Nonaka, Y., Matsukawa, N., Morohashi, K., Omura, T., Ogihara, T., Teraoka, H. & Okamoto, M. (1989) FEBS Lett. 255, 21-26] and pcP450(11 beta, aldo)-46 [Matsukawa, N., Nonaka, Y., Ying, Z., Higaki, J., Ogihara, T. & Okamoto, M. (1990) Biochem. Biophys. Res. Commun. 169, 245-252], were constructed and introduced into COS-7 cells by electroporation. Enzymatic activities of the expressed cytochromes P450(11 beta) and P450(11 beta, aldo) were determined by using 11-deoxycorticosterone, corticosterone, 18-hydroxy-11-deoxycorticosterone, 18-hydroxycorticosterone, or 19-hydroxy-11-deoxycorticosterone as a substrate. Cytochrome P450(11 beta) catalyzed 11 beta-, 18- and 19-hydroxylations of 11-deoxycorticosterone and 19-oxidation or 19-hydroxy-11-deoxycorticosterone at substantial rates, 18-hydroxylation of corticosterone at a very low rate, but no aldosterone production. Cytochrome P450(11 beta, aldo) catalyzed 11 beta- and 18-hydroxylations of 11-deoxycorticosterone, 18-hydroxylation of corticosterone and aldosterone production from 11-deoxycorticosterone or corticosterone. But neither 19-hydroxylation of 11-deoxycorticosterone nor 19-oxidation of 19-hydroxy-11-deoxycorticosterone was catalyzed by cytochrome P450(11 beta, aldo).

摘要

构建了含有大鼠细胞色素P450(11β)家族两个cDNA的表达质粒，即pcP450(11β)-62[野中洋、松川直、诸桥克彦、大村智、荻原彻、寺冈博、冈本正明(1989年)《欧洲生物化学会联合会快报》255卷，21 - 26页]和pcP450(11β,醛固酮)-46[松川直、野中洋、应志、日高纯司、荻原彻、冈本正明(1990年)《生物化学与生物物理研究通讯》169卷，245 - 252页]，并通过电穿孔法将其导入COS - 7细胞。以11 - 脱氧皮质酮、皮质酮、18 - 羟基 - 11 - 脱氧皮质酮、18 - 羟基皮质酮或19 - 羟基 - 11 - 脱氧皮质酮作为底物，测定所表达的细胞色素P450(11β)和细胞色素P450(11β,醛固酮)的酶活性。细胞色素P450(11β)能以较高速率催化11 - 脱氧皮质酮的11β - 、18 - 和19 - 羟化反应以及19 - 羟基 - 11 - 脱氧皮质酮的19 - 氧化反应，能以极低速率催化皮质酮的18 - 羟化反应，但不产生醛固酮。细胞色素P450(11β,醛固酮)能催化11 - 脱氧皮质酮的11β - 和18 - 羟化反应、皮质酮的18 - 羟化反应以及由11 - 脱氧皮质酮或皮质酮生成醛固酮。但细胞色素P450(11β,醛固酮)不催化11 - 脱氧皮质酮的19 - 羟化反应，也不催化19 - 羟基 - 11 - 脱氧皮质酮的19 - 氧化反应。

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Functional expression of the cDNAs encoding rat 11 beta-hydroxylase [cytochrome P450(11 beta)] and aldosterone synthase [cytochrome P450(11 beta, aldo)].编码大鼠11β-羟化酶[细胞色素P450(11β)]和醛固酮合酶[细胞色素P450(11β,aldo)]的cDNA的功能表达。
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