MECHANISMS OF RESPIRATION

描述：这项工作涉及酶的一般特性 与呼吸有关的。研究对象为嗜热菌 嗜热性嗜热菌。对它的生物学、生物化学有一定的实用知识 和分子生物学，已经有可能进行广泛的 其呼吸系统的生化、生物物理和物理特征 蛋白质。因为这种有机体的呼吸蛋白是 它们的功能和结构上的同源物，真核生物中的 我们的结构-功能研究结果直接适用于所有 其他类似的酶，特别是哺乳动物线粒体的酶。在.期间 过去的资助期我们：(1)确定了结构基因的DNA序列 编码细胞色素Caa3和Ba3的基因，并通过详细的比较 它们推导的氨基酸序列表明它们是 血红素/铜氧化酶超家族。(2)进行了一系列的光谱测试 细胞色素Caa3和Ba3和Rieske型蛋白的研究， 这导致了关于活动站点结构的详细新信息 这些蛋白质。(3)我们最令人兴奋的发现是 将细胞色素Ba3基因亚基II亚基进行亚克隆，在E. Coli，并形成了一个非常稳定的CUA中心。这一发现打开了 探索这部小说的性质有许多新的研究机会 以及有些出乎意料的铜原子排列，这是一个非常快的 移动的研究领域。因此，目前的提案是严格的 只关注两个科学目标，而长期目标是 项目在更广泛的技术目标下提供服务。具体目标 是：(1)为了实现对CUA结构和 函数，我们将汇编和解释光谱、结构和功能 信息来自于一种水溶性的、含有CUA的蛋白质。(2)作为第一次 为了了解CUA中心的组装，我们将研究 在CUA合成过程中形成的黄色中间体的性质。 (三)继续努力发展基因、结构相结合的 和动力学‘系统’，这将允许详细的生物物理研究 细胞色素Ba3。这项工作对人类健康的意义在于 对为所有细胞提供能量的酶有更深入的了解。那 人类呼吸蛋白的突变被惊人地发现 一个新网站的出现证明了这一点， Http://infinity.gen.emory.edu/mitomap.html.

DESCRIPTION: This work involves general characterization of enzymes involved in respiration. The subject organism is the thermophilic bacterium Thermus thermophilus. With a working knowledge of its biology, biochemistry and molecular biology, it has been possible to carry out a wide range of biochemical, biophysical, and physical characterizations of its respiratory proteins. Because the respiratory proteins from this organism are functional and structural homologs of their eukaryotic counterparts, the results of our structure-function studies are directly applicable to all other such enzymes, notably those of the mammalian mitochondrion. During the past funding period we: (1) Determined the DNA sequence of structural genes encoding cytochromes caa3 and ba3, and by making detailed comparisons of their deduced amino acid sequences demonstrated that they are members of heme/Cu oxidase super-family. (2) We carried out a number of spectroscopic investigations with cytochromes caa3 and ba3 and of Rieske-type proteins, which led to detailed new information about the active site structures of these proteins. (3) Our most exciting discovery was that a small portion of the subunit II of cytochrome ba3 gene could be subcloned, expressed in E. coli, and was found to form a very stable CuA center. This discovery opens many new research opportunites for exploring the properties of this novel and somewhat unexpected arrangement of Cu atoms, and this is a very fast moving field of research. Accordingly, the current proposal is tightly focussed on just two scientific goals while the longer-term goals of the project are served under a much broader technical goal. The Specific Aims are: (1) Toward achieving a detailed understanding of CuA structure and function, we will assemble and interpret spectral, structural and functional information from a water-soluble, CuA containing protein. (2) As a first step toward understanding assembly of the CuA center, we will examine the properties of a yellow-colored intermediate that forms during CuA synthesis. (3) We will continue our efforts to develop a combined genetic, structural and kinetic 'system' that will permit detailed biophysical study of the cytochrome ba3. The significance of this work to human health lies in a deeper understanding of the enzymes that provide energy to all cells. That mutations in human respiratory proteins are being found at a prodigious rate, is evidenced by the appearance of a new web site, http://infinity.gen.emory.edu/mitomap.html.