HEART MUSCLE AND BACTERIAL RESPIRATORY CHAIN SYSTEMS

我们的目标是更深入地了解心脏的电子传输 通过研究和比较心脏线粒体和一种细菌， 与哺乳动物线粒体的关系。 我们将调查 哺乳动物心脏线粒体内膜呼吸链 肌肉和细胞质膜的好氧细菌副球菌 使用我们开发的技术。 重点抓好 细胞色素c的反应的功能和结构方面 将电子从细胞色素c还原酶传递到细胞色素c氧化酶。 我们将继续研究形成我们的 一种新的、高周转率的、特别是活性的细胞色素c-细胞色素复合物 氧化酶，其具有增加的组分之间的相互作用，或 更快速还原或氧化适当组分的能力。 我们将检查所需的实验条件， 细胞色素c-细胞色素c类似反应性复合物的出现 还原酶。 由于细胞色素氧化酶的测定方法多种多样， 不同的实验室所采用的方法， 如果我们的决定不具有可比性或可解释性，我们将努力建立一个 分光光度法与极谱法统一验收关联法 测定。 我们的实验室是第一个开发出单克隆抗体， 细胞色素c。 使用我们已经产生的杂交瘤细胞系， 产生了15种不同的特异性单克隆抗体， 牛肉细胞色素c和P. 其中一些 抗体仅结合同源抗原，并且一些是交叉反应性的。 我们将利用这个单克隆抗体库， 继续开发，以探测细胞色素c上的位点， 参与其与氧化酶，还原酶和过氧化物酶的相互作用， 哺乳动物和细菌的电子传递系统。 我们将执行 确定不同表位数目的竞争结合测定 （抗原位点）被这些同质抗体识别。 研究将 针对的是P. Escherificans的电子传递链， 特别感兴趣的是，因为它是如此相似的组成链 线粒体内膜，并可能是其进化的前体。 这种生物体的其他优点是亚细胞 颗粒可以被分离，并且可以改变颗粒的组分， 改变生长条件。

Our objective is to gain deeper insight into electron transport in heart muscle by studying and comparing heart mitochondria and a bacterium chosen for its relationship to mammalian mitochondria. We will investigate the respiratory chain of the inner mitochondrial membrane of mammalian heart muscle and the cytoplasmic membrane of the aerobic bacterium Paracoccus denitrificans with techniques we have developed. We will focus on the functional and structural aspects of the reactions by which cytochrome c transports electrons from cytochrome c reductase to cytochrone c oxidase. We will continue to study the conditions needed for the formation of our new, high-turnover, especially reactive, complex of cytochrome c-cytochrome oxidase that has either increased interaction between the components or the capacity for more rapid reduction or oxidation of the proper components. We will examine the experimental conditions which are required for the appearance of a similar reactive complex of cytochrome c-cytochrome c reductase. Since the variety of assay procedures for cytochrome oxidase that have been employed by different laboratories have resulted in determinations not comparable or interpretable, we will work to establish a uniform acceptable method of relating spectrophotometric and polarographic assays. Our laboratory is the first to develop monoclonal antibodies to cytochrome c. Using hybridoma cell lines that we have generated, we have produced 15 different exquisitely specific monoclonal antibodies to both beef cytochrome c and P. denitrificans cytochrome c. Some of these antibodies bind only the homologous antigen and some are cross reactive. We will utilize this library of monoclonal antibodies which we are continuing to develop, to probe sites on the cytochrome c which are involved in its interactions with oxidases, reductases and peroxidases in mammalian and bacterial electron transport systems. We will perform competition binding assays to determine the number of distinct epitopes (antigenic sites) recognized by these homogeneous antibodies. Studies will be aimed at the electron transport chain of P. denitrificans which is of particular interest because it is so similar in composition to the chain of the mitochondrial inner membrane, and may be its evolutionary precursor. Other advantages of this organism are the ease with which subcellular particles can be isolated, and the possibility of varying components of the chain by changing the growth conditions.