Section of Biochemistry and Molecular Biology, Cornell University, Ithaca, New York 14850.
J Gen Physiol. 1969 Jul 1;54(1):38-49. doi: 10.1085/jgp.54.1.38.
The problem of the resolution and reconstitution of the inner mitochondrial membrane has been approached at three levels. (1) Starting with phosphorylating submitochondrial particles, a "resolution from without" can be achieved by stripping of surface components. The most extensive resolution was recently obtained with the aid of silicotungstate. Such particles require for oxidative phosphorylation the addition of several coupling factors as well as succinate dehydrogenase. (2) Starting with submitochondrial particles that have been degraded by trypsin and urea a resolution of the inner membrane proper containing an ATPase has been achieved. These experiments show that at least five components are required for the reconstitution of an oligomycin-sensitive ATPase: a particulate component, F(1), Mg(++), phospholipids, and F(c). Morphologically, the reconstituted ATPase preparations resemble submitochondrial particles. (3) Starting with intact mitochondria individual components of the oxidation chain have been separated from each other. The following components were required for the reconstitution of succinoxidase: succinate dehydrogenase, cytochrome b, cytochrome c(1), cytochrome c, cytochrome oxidase, phospholipids and Q(10). The reconstituted complex had properties similar to those of phosphorylating submitochondrial particles; i.e., the oxidation of succinate by molecular oxygen was highly sensitive to antimycin.
这个问题的决议和重建的内部线粒体膜已经在三个层面上。(1)从磷酸化的亚线粒体颗粒开始，可以通过去除表面成分来实现“从外部解析”。最近，在硅钨酸盐的帮助下，获得了最广泛的解析。这种颗粒需要添加几个偶联因子以及琥珀酸脱氢酶才能进行氧化磷酸化。(2)从已被胰蛋白酶和尿素降解的亚线粒体颗粒开始，已经实现了含有 ATP 酶的内膜的解析。这些实验表明，至少需要五个成分才能重建寡霉素敏感的 ATP 酶：颗粒成分 F(1)、Mg(++)、磷脂和 F(c)。形态学上，重建的 ATP 酶制剂类似于亚线粒体颗粒。(3)从完整的线粒体开始，氧化链的各个成分已经彼此分离。以下成分是重新构建琥珀酸氧化酶所必需的：琥珀酸脱氢酶、细胞色素 b、细胞色素 c(1)、细胞色素 c、细胞色素氧化酶、磷脂和 Q(10)。重建的复合物具有类似于磷酸化亚线粒体颗粒的性质；即，分子氧对琥珀酸的氧化高度敏感于抗霉素。
