BIOENERGETIC APPARATUS IN HEART MITOCHONDRIA

这项研究的目的是处理非细胞色素 组分是为了获得呼吸链酶的基本信息 线粒体内膜系统，从而使结论 关于强制性承运人、其顺序、行动、机制和 与电子传输链有关的主题，以及确定 膜在这些过程中的作用，利用 顺序分解、系统重构、识别和 分离出的酶的特性。首先，分解的酶和 组件将被提纯，然后由所有 可想象的物理化学和生化技术，包括反应 动力学、氨基酸序列、EPR、Endor、结晶、EXAFS和 萨尼斯等人。氨基酸序列测定是最具确定性的 在描述组件的身份或非身份时。然而，它 必须强调的是，重建将被用作最终和最 一种可靠的方法，以确保在以下过程中未发生降级或更改 对成分进行广泛的提纯。同时，有目的的 根据合理的设计更改或修改隔离的 将使用酶来检测重组活性，以便 确定性质允许偏离正常的回旋余地有多大 功能。然而，最终的目标是从明确的定义中重建 具有所有功能的完整电子转移装置的部件 线粒体的表现。 应该指出的是，此应用程序(与另一应用程序不同 项目)强调并处理不含细胞色素的 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.