PROTON & ELECTRON TRANSFER & ENERGY COUPLING

该项目的最终目标是阐明结构- I位和III位能量耦合的函数关系。 我们 有三个主要的长期目标，即（A）测试 假设位点I中的电子转移机制是 一个循环的类型。 这一假设使我们能够解释（a） 氧化还原组分和蛋白质亚基的极端复杂性 结构，（B）高（四至五）化学计量的H+/2 e-（c） 极低的中间电位的集群N-la。 为了实现这一目标，我们将做以下工作：（一）研究物理- H+载体的化学性质，（ii）研究新的 与传统鱼藤酮型不同的抑制剂 化合物，（iii）测量氧化还原的空间组织 使用连续波和脉冲波测量膜中的组分 EPR技术在低温下，和（iv）研究更简单 细菌系统，如嗜热副球菌和栖热菌 嗜热菌HB-8。 (B)将我们在研究中心I的研究经验应用于临床 问题 某些疾病是由基因缺陷引起的， 铁硫蛋白在站点I. 我们已经证明， 缺陷可以通过EPR测量清楚地识别， 病理标本 我们将继续这方面的研究。 (C)阐明位点III的结构功能关系 呼吸 自从我们率先测量自旋-自旋 cyt.a3-NO和其他顺磁性物质之间的相互作用 牛心脏细胞色素氧化酶，我们将完成这项研究， 进行以下实验：（i）确定性测量 牛心脏细胞色素氧化酶的簇间距离，（ii） 通过脉冲EPR直接测量T1和T2， cyt.a和CuA的氧化还原平衡的解析，（iii）酵母菌株的使用 谁的Cyt c氧化酶蛋白和发色团结构是 受到双重突变的干扰 充分利用学习 微生物，我们将研究光谱和热力学 单个氧化还原组分的特性和对 氧化还原和氧化还原中心之间的自旋-自旋相互作用。 这些研究有望为我们的研究开辟新的视野。 理解Site I中的电子和质子转移过程 和III在分子水平上。

The ultimate goal of the project is to elucidate the structure- function relationship of the Site I and Site III energy coupling. We have three major long term objectives, namely, (A) To test the hypothesis that the mechanism of electron transfer in the Site I is of a cyclic type. This hypothesis enables us to explain (a) the extreme complexity of redox components and protein subunit structure, (b) a high (four to five) stoichiometry of H+/2e- (c) the extremely low midpotential of the cluster N-la. For this goal, we will do the following: (i) study the physico- chemical properties of the H+-carriers, (ii) investigate new inhibitors which are different from conventional rotenone-type compounds, (iii) measure the spatial organization of the redox components in the membrane using continuous wave and pulsed EPR techniques at cryogenic temperature, and (iv) study simpler bacterial systems, such as Paracoccus denitrificans and Thermus thermophilus HB-8. (B) To apply our experience in the study of Site I to clinical problems. Certain diseases are caused by a genetic defect in the iron sulfur proteins in the Site I. We have demonstrated that this defect can be clearly identified by EPR measurement on pathological specimens. We will continue this line of study. (C) To elucidate the structure function relationship of Site III respiration. Since we pioneered the measurement of spin-spin interaction between cyt.a3-NO and other paramagnetic species in bovine heart cytochrome oxidase, we will complete the study by conducting the following experiments: (i) definitive measurement of inter-cluster distance in bovine heart cytochrome oxidase, (ii) direct measurement of T1 and T2 by pulsed EPR with careful resolution of redox poise of cyt.a and CuA, (iii) use of yeast strain whose cyt. c oxidase protein and chromophore structure are perturbed by double mutation. Taking full advantage of studying microorganism, we will study spectral and thermodynamic characteristics of individual redox components and analysis of the redox and spin-spin interactions among redox centers. These studies are expected to open a new horizon in our understanding of the electron and proton transfer process in Site I and III on the molecular level.