RECONSTITUTION OF MITOCHONDRIAL RESPIRATORY CHAIN

这项研究的目的是获得有关呼吸道疾病的基本信息 线粒体内膜的链酶系统使能结论 电子-氢的载流子及其序列、作用和机理 运输(最终导致能量耦合)。计划中的方法 将主要通过顺序拆分、系统重构、酶 身份识别和相关技术。其中，分解的酶 首先对成分进行提纯(如果可能的话，最好是结晶) 然后通过各种理化和酶的严格表征 技术，如快速和慢速动力学方法，氨基酸 序列分析、磁共振技术、EXAFS等。 强调重建将被用作最可靠的技术 (目前)以确定未发生变性或变化 为了获得明确的结果，对成分进行了广泛的提纯。 我们第一阶段的最终目标，不一定是最终目标，是 具有明确定义的呼吸酶或其成分的重组 构成生物能量学一部分的完整的电子传输链 具有线粒体所有功能表现的器官。 必须指出，此应用程序(与另一应用程序不同 项目)强调并处理含有细胞色素的部件的各方面 除了与呼吸链最密切相关的成分外 细胞色素。

The aim of this research is to obtain basic information on the respiratory chain enzyme system of mitochondrial inner membrane enabling conclusions on carriers, their sequence, actions and mechanisms of electron-hydrogen transport (which eventually leads to energy coupling). The methods planned will be mainly by sequential resolution, systematic reconstitution, enzyme identification, and related techniques. Among others, the resolved enzymes and components are first purified (preferably crystallized, if possible) and then rigorously characterized by various physicochemical and enzymic techniques, such as rapid and "slow" methods for kinetics, amino acid sequence analysis, magnetic resonance techniques, EXAFS, etc. It should be emphasized that reconstitution will be used as a most reliable technique (at present) to ascertain that no denaturation or changes occurred in the extensive purification of components in order to obtain clear-cut results. An ultimate goal in our first phase, not necessarily the eventual one, is reconstitution with well defined respiratory enzymes or components of the complete electron transport chain which constitutes a part of bioenergetic apparatus with all functional manifestations of the mitochondrion. It must be pointed out that this application (which differs from another project) emphasizes and deals with aspects on cytochrome-containing parts of the respiratory chain, except components most intimately associated with cytochromes.