ELECTRON AND PROTON TRANSLOCATION IN BIOLOGICAL ENERGY

生物能中的电子和质子易位

• 批准号: 2701499
• 负责人: PETER LESLIE DUTTON
• 金额: $ 34.05万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 1979
• 资助国家: 美国
• 起止时间: 1979-05-01 至 1999-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2701499
• 关键词: Rhodopseudomonas; X ray crystallography; acidity /alkalinity; bioenergetics; biological transport; chemical binding; chromophore; cofactor; cytochrome b; cytochrome c; cytochrome oxidase; electrochemistry; electron transport; enzyme complex; enzyme electrodes; enzyme mechanism; enzyme structure; hemoprotein structure; high performance liquid chromatography; hydrogen transport; oxidation reduction reaction; photosynthesis; quinones; site directed mutagenesis; surface property; ubiquinone

We focus on mechanisms that promote energy conserving electron and proton translocation through ubiquinol-cytochrome c oxidoreductase (bc1 complex). We shall make use of experimental systems that offer the means to activate the system by light flashes via a on a bacterial photochemical reaction center. Such systems can provide valuable information on a rapid timescale, of kinetics of oxidation/reduction and protonation/deprotonation of ratios of electron and proton movements and, in combination with redox potentiometry, of the fundamental properties of the active redox components. The first part is aimed at the mechanism of action of bc1 complexes in vivo and in vitro. The principal systems are: a) the native membranes from photosynthetic bacteria (Rhodopseudomonas sphaeroides and capsulata); b) hybrid constructions in vitro of well-characterized isolated reaction centers (Rps. sphaeroides), cytochrome c and bc1 complexes, (beef heart mitochondria). The in vivo photosynthetic bacterial membranes provide a coupled photon-electron-proton-phosphorylation membrane system that possesses the usual manipulative opportunities offered by bacteria, including growth/media manipulations and mutants blocked in the bc1, complex. The in vitro hybrid constructions provide wide experimental flexibility and provide the means of achieving more ordered, densely packed mono- and multilayer arrays of the hybrid systems on glass or electrodes, for structurally related spectrometric analysis and direct measurement and control of charge movements across the monolayer profile. The second part approaches the bc1 complex from a different direction: from the study of a key component--ubiquinone. Quinonoid compounds will be studied from the standpoint of their structure, their substituents, electrochemistry, and hydrophobic properties. Quinonoid compounds will be used to probe the bc1 complex catalytic sites of ubiquinone oxidation-reduction to determine the parameters that govern the association of molecule and reaction site(s). Further efforts will be made to develop a family of bc complexes, systematically altered in the electrochemistry of the replacement quinones. This approach should strongly augment features of the first theme of the proposal. Moreover, it will provide the basis for a much needed, alternate approach to obtaining kinetic and thermodynamic information from the systematically altering free energy gaps between reactants and add another dimension to the study of electric field effects on the bc1 complexes oriented as mono- and multilayers between electrode surfaces.

我们专注于促进能源保存电子和质子的机制 通过泛素醇 - 卵色素C氧化还原酶（BC1复合物）的易位。 我们将利用提供激活手段的实验系统 光线通过细菌光化学反应闪烁的系统闪烁 中心。 这样的系统可以快速提供有价值的信息 时间尺度，氧化/还原和质子化/去质子化的动力学 电子和质子运动的比率，并与氧化还原结合 电位测定法，活性氧化还原成分的基本特性。 第一部分针对体内BC1复合物的作用机理 并在体外。 主要系统是：a）来自 光合细菌（Rhodopseudomonas sphaeroides和capsulata）； b） 特征良好的分离反应的体外杂种构造 中心（Rps。Sphaeroides），细胞色素C和BC1配合物，（牛肉心脏 线粒体）。 体内光合细菌膜提供 耦合的光子电子 - 普罗顿 - 磷酸化膜系统 拥有细菌提供的通常的操纵机会， 包括在BC1中阻塞的增长/媒体操纵和突变体 复杂的。 体外杂交结构提供广泛的实验 灵活性并提供实现更多有序，密集的填充的手段 玻璃或电极上混合系统的单层和多层阵列， 用于结构相关的光谱分析和直接测量以及 控制跨单层轮廓的电荷运动。 第二部分从不同的方向接近BC1复合物： 根据关键成分的研究 - 葡萄类酮。 喹酮化合物将是 从结构的角度研究，取代基 电化学和疏水性质。 喹酮化合物将是 用于探测泛氨基酮的BC1复合物催化位点 减少氧化 - 确定控制关联的参数 分子和反应位点（S）。 将采取进一步的努力来发展 卑诗省综合体的家族，有系统地改变了 替换奎因酮。 这种方法应强烈增加功能 提案的第一个主题。 而且，它将提供基础 对于急需的，替代方法来获得动力学和 来自系统改变自由能差距的热力学信息 在反应物之间并为电场的研究增加另一个维度 对以单层为单层和多层的BC1络合物的影响 电极表面。

项目成果

A new method of solvation analysis: applications to quinones.

溶剂化分析的新方法：在醌中的应用。

• DOI: 10.1515/znc-1990-0520
• 发表时间: 1990
• 期刊: Zeitschrift fur Naturforschung. C, Journal of biosciences
• 作者: Keske,JM;Bruce,JM;Dutton,PL
• 通讯作者: Dutton,PL

The nature and magnitude of the charge-separation reactions of ubiquinol cytochrome c2 oxidoreductase.

泛醇细胞色素 c2 氧化还原酶的电荷分离反应的性质和强度。

• DOI: 10.1016/0005-2728(88)90223-x
• 发表时间: 1988
• 期刊: Biochimica et biophysica acta
• 作者: Robertson,DE;Dutton,PL
• 通讯作者: Dutton,PL

The Rhodospirillum rubrum cytochrome bc1 complex: redox properties, inhibitor sensitivity and proton pumping.

红色红螺菌细胞色素 bc1 复合物：氧化还原特性、抑制剂敏感性和质子泵送。

• DOI: 10.1016/s0005-2728(05)80247-6
• 发表时间: 1991
• 期刊: Biochimica et biophysica acta
• 作者: Güner,S;Robertson,DE;Yu,L;Qiu,ZH;Yu,CA;Knaff,DB
• 通讯作者: Knaff,DB

Experimental resolution of the free energies of aqueous solvation contributions to ligand-protein binding: quinone-QA site interactions in the photosynthetic reaction center protein.

水溶剂化自由能对配体-蛋白质结合贡献的实验分辨率：光合反应中心蛋白质中的醌-QA 位点相互作用。

• DOI: 10.1073/pnas.90.7.2920
• 发表时间: 1993
• 期刊: Proceedings of the National Academy of Sciences of the United States of America
• 影响因子: 11.1
• 作者: Warncke,K;Dutton,PL
• 通讯作者: Dutton,PL

The reaction center profile structure derived from neutron diffraction.

由中子衍射得出的反应中心轮廓结构。

• DOI: 10.1016/0005-2728(81)90026-8
• 发表时间: 1981
• 期刊: Biochimica et biophysica acta
• 作者: Pachence,JM;Dutton,PL;Blasie,JK
• 通讯作者: Blasie,JK