MECHANISMS OF RESPIRATION

描述：这项工作涉及酶的一般特性 参与呼吸。 目标生物是嗜热菌 栖热菌 有了它的生物学，生物化学， 和分子生物学，已经有可能进行广泛的 其呼吸系统的生物化学、生物物理和物理特征 proteins. 因为这种生物的呼吸蛋白 它们与真核生物的功能和结构同源物， 我们的结构-功能研究结果直接适用于所有 其他这类酶，特别是哺乳动物的胰蛋白酶。 期间 在过去的资助期间，我们：（1）确定了结构的DNA序列 编码细胞色素caa 3和ba 3的基因，并进行详细的比较 其推导的氨基酸序列表明，它们是 血红素/铜氧化酶超家族。 (2)我们进行了大量的光谱分析 用细胞色素caa 3和ba 3以及Rieske型蛋白质的研究， 这导致了关于活性位点结构的详细新信息， 这些蛋白质。 (3)我们最激动人心的发现是 细胞色素ba 3亚基II基因可在大肠杆菌中亚克隆表达。 大肠杆菌中，并发现形成一个非常稳定的CuA中心。 这一发现开启了 许多新的研究机会来探索这部小说的性质 和一些意想不到的铜原子排列，这是一个非常快的 移动研究领域 因此，目前的建议是严格的， 只专注于两个科学目标，而长期目标的 项目是在更广泛的技术目标下服务的。 具体目标 （1）对CuA结构的详细了解， 功能，我们将组装和解释光谱，结构和功能 来自水溶性的含有CuA的蛋白质的信息。 (2)作为第一 为了进一步了解CuA中心的组装，我们将研究 CuA合成过程中形成的黄色中间体的性质。 (3)我们将继续努力开发一种结合基因、结构 和动力学'系统'，这将允许详细的生物物理研究， 细胞色素ba 3。 这项工作对人类健康的意义在于 更深入地了解为所有细胞提供能量的酶。 的 人类呼吸道蛋白质的突变被发现， 率，是证明了一个新的网站的外观， 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.