Cell-free synthesis of precursor forms of mitochondrial steroid hydroxylase enzymes of the bovine adrenal cortex.
Cell-free synthesis of precursor forms of mitochondrial steroid hydroxylase enzymes of the bovine adrenal cortex.
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牛肾上腺皮质线粒体类固醇羟化酶前体形式的无细胞合成。
DOI：
10.1016/0003-9861(82)90106-0
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发表时间：
1982
影响因子：
3.9
通讯作者：
Waterman,MR
中科院分区：
文献类型：
--
作者：
Kramer,RE;DuBois,RN;Simpson,ER;Anderson,CM;Kashiwagi,K;Lambeth,JD;Jefcoate,CR;Waterman,MR
从放射性标记的牛肾上腺皮质细胞和用牛肾上腺皮质RNA编程的兔网织红细胞裂解物翻译系统中免疫分离新合成的[35 S]甲硫氨酸标记的胆固醇侧链裂解细胞色素P-450、11β-羟化酶细胞色素P-450、肾上腺素还蛋白和肾上腺素还蛋白还原酶。从网织红细胞裂解物翻译系统免疫分离的胆固醇侧链裂解细胞色素P-450的表观分子量为54，500，而从放射性标记的牛肾上腺皮质细胞免疫分离的细胞色素P-450的表观分子量为49，000，与纯化蛋白的表观分子量相同。同样，从网织红细胞裂解物翻译系统免疫分离的新合成的[35 S]甲硫氨酸标记的11β-羟化酶细胞色素P-450的表观分子量比从放射性标记的肾上腺皮质细胞（48，000）和真实细胞色素（48，000）免疫分离的表观分子量大5500道尔顿。肾上腺素还蛋白和肾上腺素还蛋白还原酶的无细胞翻译产物也比相应的线粒体蛋白大几千道尔顿。从网织红细胞裂解物翻译系统中免疫分离的肾上腺素还蛋白的表观分子量为19，000，而真实蛋白的表观分子量为12，000。免疫分离的溶胞产物翻译系统的肾上腺素还蛋白还原酶的表观分子量为53，400;比免疫分离的放射性标记的肾上腺皮质细胞或通过常规技术纯化的肾上腺素还蛋白还原酶的表观分子量大2300道尔顿。这些结果表明，牛肾上腺皮质的线粒体类固醇羟化酶系统的所有组分都是作为较高分子量的前体分子合成的。据推测，前体蛋白质在插入到外显子中时通过包括前体片段的蛋白水解切割的过程后水解转化为成熟酶。
Newly synthesized, [35S]methionine-labeled cholesterol side-chain cleavage cytochromeP-450, 11β-hydroxylase cytochromeP-450, adrenodoxin, and adrenodoxin reductase were immunoisolated from radiolabeled bovine adrenocortical cells and from rabbit reticulocyte lysate translation systems programmed with bovine adrenocortical RNA. Cholesterol side-chain cleavage cytochromeP-450 immunoisolated from a reticulocyte lysate translation system had an apparent molecular weight of 54,500 whereas this cytochromeP-450 immunoisolated from radiolabeled bovine adrenocortical cells had an apparent molecular weight of 49,000, an apparent molecular weight identical to that of the purified protein. Similarly, newly synthesized, [35S]methionine-labeled 11β-hydroxylase cytochromeP-450 immunoisolated from a reticulocyte lysate translation system had an apparent molecular weight 5500 daltons larger than that immunoisolated from radiolabeled adrenocortical cells (48,000) and the authentic cytochrome (48,000). The cell-free translation products of adrenodoxin and adrenodoxin reductase were also several thousand daltons larger than the corresponding mitochondrial proteins. The apparent molecular weight of adrenodoxin immunoisolated from a reticulocyte lysate translation system was 19,000, while that of the authentic protein was 12,000. Adrenodoxin reductase immunoisolated from a lysate translation system had an apparent molecular weight of 53,400; an apparent molecular weight 2300 daltons larger than that of adrenodoxin reductase immunoisolated from radiolabeled adrenocortical cells or purified by conventional techniques. These results demonstrate that all of the components of the mitochondrial steroid hydroxylase systems of the bovine adrenal cortex are synthesized as precursor molecules of higher molecular weight. Presumably, the precursor proteins are post-translationally converted to the mature enzymes upon insertion into the mitochondrion by a process which includes the proteolytic cleavage of the precursor segments.
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影响因子：
4.8
作者：
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DOI：
--
发表时间：
1981
期刊：
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Waterman,MR
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发表时间：
1967-10
期刊：
European journal of biochemistry
影响因子：
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作者：
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影响因子：
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作者：
Nagasumi Yago;Shigeru Kobayashi;Shigetaka Sekiyama;Hiromi Kurokawa;Yuko Iwai;Ichiro Suzuki;Shogo Ichii
通讯作者：
Shogo Ichii
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DOI：
10.1016/s0021-9258(19)43850-7
发表时间：
1980
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
E. Schmelzer;P. Heinrich
通讯作者：
P. Heinrich