Madden T D, Cullis P R
J Biol Chem. 1984 Jun 25;259(12):7655-8.
A preparation of reconstituted cytochrome oxidase vesicles in which the enzyme is oriented facing inwards (such that it cannot interact with external cytochrome c) is described. No oxidase activity is expressed by these vesicles unless they are disrupted, allowing influx of cytochrome c or exposure of the oxidase-binding site to the external medium. We have exploited this property to follow detergent-induced solubilization of the membrane, a technique which allows membrane disruption and enzyme activity to be monitored simultaneously. This protocol can be employed to investigate the properties and mechanism of action of detergents as is illustrated for several ionic and nonionic detergents.
本文描述了一种重组细胞色素氧化酶囊泡制剂，其中酶的方向是向内（使其不能与外部细胞色素c相互作用）。除非这些囊泡被破坏，使细胞色素c流入或氧化酶结合位点暴露于外部介质，否则它们不会表现出氧化酶活性。我们利用这一特性来跟踪去污剂诱导的膜溶解，该技术允许同时监测膜破坏和酶活性。正如对几种离子型和非离子型去污剂所说明的那样，该方案可用于研究去污剂的性质和作用机制。
