RECONSTITUTION OF MITOCHONDRIAL RESPIRATORY CHAIN

本研究的目的是获得呼吸系统的基本信息， 链酶系统的线粒体内膜使结论 电子氢的载流子、顺序、作用和机理 运输（最终导致能量耦合）。 计划的方法 将主要通过顺序拆分、系统重组、酶 识别和相关技术。 其中，分解的酶 并且组分首先被纯化（如果可能，优选结晶）， 然后通过各种物理化学和酶的严格表征， 技术，如快速和“慢”的动力学方法，氨基酸 序列分析，磁共振技术，EXAFS等，它应该是 强调重组将被用作最可靠的技术 (at存在），以确定未发生变性或变化， 对组分进行广泛纯化，以获得明确的结果。 我们第一阶段的最终目标，不一定是最终目标，是 用定义明确的呼吸酶或呼吸酶的组分进行重构， 完整的电子传递链，构成生物能量的一部分 这是一个具有所有功能表现的装置。 必须指出的是，本申请（其不同于另一个申请）是在本申请的范围内进行的。 项目）强调和处理的方面，细胞色素包含的部分 呼吸链，除了最密切相关的成分， 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.