Structure and orientation of cytochrome c oxidase in crystalline membranes. Studies by electron microscopy and by labeling with subunit-specific antibodies.
Structure and orientation of cytochrome c oxidase in crystalline membranes. Studies by electron microscopy and by labeling with subunit-specific antibodies.
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晶体膜中细胞色素 C 氧化酶的结构和方向。
DOI：
10.1016/s0021-9258(17)34732-4
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发表时间：
1978
期刊：
影响因子：
--
通讯作者：
G. Schatz
中科院分区：
文献类型：
--
作者：
T. Frey;S. Chan;G. Schatz
结晶细胞色素c氧化酶膜中细胞色素c氧化酶分子的结构和取向（Vanderkooi，G.，老艾Capaldi，R.A.，和Hayashi，H.（1972）Biochim. Biophys. Acta 274，38-48）通过电子显微照片的图像分析和通过使结晶制剂与针对单个细胞色素c氧化酶亚基的免疫γ-球蛋白反应来研究。通过以下两种方法检测γ-球蛋白与膜的结合：（a）电泳鉴定洗涤膜中的γ-球蛋白多肽;（B）电子显微镜检查负染色膜。膜结合免疫γ-球蛋白对抗亚基IV（其在完整线粒体中面向基质侧），但未能结合免疫γ-球蛋白对抗亚基II + III（其在完整线粒体中面向内膜外侧）。与此相反，溶解的细胞色素c氧化酶绑定的两个免疫γ-球蛋白。因此，所有细胞色素c氧化酶分子在晶体膜中不对称地排列，使得亚基IV面向外，亚基II + III面向内。这种取向与完整线粒体的取向相反。这些数据还表明，结晶膜形成封闭的囊泡，外部添加的γ-球蛋白是不可渗透的。
The structure and the orientation of cytochrome c oxidase molecules in crystalline cytochrome c oxidase membranes (Vanderkooi, G., Senior, A.E., Capaldi, R.A., and Hayashi, H. (1972) Biochim. Biophys. Acta 274, 38-48) were studied by image analysis of electron micrographs and by reacting the crystalline preparations with immune gamma-globulins against individual cytochrome c oxidase subunits. Binding of gamma-globulins to the membranes was detected by the following two methods: (a) electrophoretic identification of gamma-globulin polypeptides in the washed membranes; (b) electron microscopic examination of the negatively stained membranes. The membranes bound immune gamma-globulins against subunit IV (which faces the matrix side in intact mitochondria) but failed to bind immune gamma-globulins against subunits II + III (which face the outer side of the inner membrane in intact mitochondria). In contrast, solubilized cytochrome c oxidase bound either of the two immune gamma-globulins. All cytochrome c oxidase molecules in the crystalline membranes are thus asymmetrically arranged so that subunit IV faces outward and subunits II + III face toward the interior. This orientation is opposite to that found with intact mitochondria. The data also suggest that the crystalline membranes form closed vesicles which are impermeable to externally added gamma-globulins.