细胞色素P450侧链裂解酶-肾上腺皮质铁氧化还原蛋白还原酶-肾上腺皮质铁氧化还原蛋白融合蛋白导入酵母线粒体中催化结构域的相互作用

Interaction of catalytic domains in cytochrome P450scc--adrenodoxin reductase--adrenodoxin fusion protein imported into yeast mitochondria.

作者信息

Novikova L A, Nazarov P A, Saveliev A S, Drutsa V L, Sergeev V N, Miller W L, Luzikov V N

机构信息

Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University, Moscow, 119899 Russia.

出版信息

Biochemistry (Mosc). 2000 Dec;65(12):1362-6. doi: 10.1023/a:1002844604728.

DOI:10.1023/a:1002844604728

PMID:11173506

Abstract

We have constructed plasmids for yeast expression of the fusion protein pre-cytochrome P450scc--adrenodoxin reductase-adrenodoxin (F2) and a variant of F2 with the yeast CoxIV targeting presequence. Mitochondria isolated from transformed yeast cells contained the F2 fusion protein at about 0.5% of total protein and showed cholesterol hydroxylase activity with 22(R)-hydroxycholesterol. The activity increased 17- or 25-fold when sonicated mitochondria were supplemented with an excess of purified P450scc or a mixture of adrenodoxin (Adx) and adrenodoxin reductase (AdxRed), respectively. These data suggest that, at least in yeast mitochondria, the interactions of the catalytic domains of P450scc, Adx, and AdxRed in the common polypeptide chain are restricted.

摘要

我们构建了用于酵母表达融合蛋白前细胞色素P450scc-肾上腺odoxin还原酶-肾上腺odoxin（F2）以及带有酵母CoxIV靶向前序列的F2变体的质粒。从转化的酵母细胞中分离出的线粒体含有占总蛋白约0.5%的F2融合蛋白，并对22(R)-羟基胆固醇表现出胆固醇羟化酶活性。当超声处理的线粒体分别补充过量的纯化P450scc或肾上腺odoxin（Adx）与肾上腺odoxin还原酶（AdxRed）的混合物时，活性分别增加了17倍或25倍。这些数据表明，至少在酵母线粒体中，P450scc、Adx和AdxRed的催化结构域在共同多肽链中的相互作用受到限制。