BIOENERGETIC APPARATUS IN HEART MITOCHONDRIA

这项研究的目的是处理不含细胞色素的 组件是为了获得呼吸链酶的基本信息 线粒体内膜系统，以便能够得出结论 关于强制承运人的规定，其顺序、行动、机制和 与电子传递链相关的主题，以及确定 膜在这些过程中的作用，利用的方法， 顺序解析、系统重构、识别和 分离的酶的表征。 首先，分解的酶和 成分将被纯化，然后通过所有 可以想象的物理化学和生物化学技术，包括反应 动力学，氨基酸序列，EPR，ENDOR，结晶，EXAFS和 除此之外，SANS。 氨基酸序列测定是最确定的 在描述组件的同一性或非同一性时。 但 必须强调的是，重组将被用作最终和最重要的 可靠的方法，以确定没有退化或变化发生在 组分的广泛纯化。 与此同时， 基于合理的设计，对隔离的 酶将用于检查重建活性， 确定自然界允许偏离正常值的余地 功能 然而，最终目标是从定义明确的 具有所有功能的完整电子转移装置的组件 的表现形式。 应该指出的是，本申请（其与另一个申请不同） 项目）强调和处理非细胞色素的各个方面， 呼吸链的组成部分。

The aims of this research which deals with non-cytochrome-containing components are to obtain basic information on the respiratory chain enzyme system of mitochondrial inner membrane so as to enable conclusions concerning obligatory carriers, their sequence, actions, mechanisms and subjects related to the electron transport chain, as well as to determine the role of the membrane in these processes, utilizing the methods of sequential resolution, systematic reconstitution, identification and characterization of the isolated enzymes. First, the resolved enzymes and components will be purified and then rigorously characterized by all conceivable physico-chemical and biochemical techniques, including reaction kinetics, amino acid sequence, EPR, ENDOR, crystallization, EXAFS and SANES, among others. Amino acid sequence determination is most definitive in delineating the identity or non-identity of the components. However, it must be emphasized that reconstitution will be used as the final and most reliable method to ascertain that no degradation or changes occurred during extensive purification of the components. At the same time, purposeful change or modification, based on well-reasoned design, of the isolated enzymes will be used to examine the reconstitutive activity in order to ascertain how much leeway nature allows for deviation from normal function. However, an ultimate goal is reconstitution from well defined components of the complete electron transfer apparatus with all functional manifestations of the mitochondrion. It should be pointed out that this application (which differs from another project) emphasizes and deals with aspects of the non-cytochrome-containing components of the respiratory chain.