Natural engineering of multi-electron biological oxidation & reduction

多电子生物氧化自然工程

• 批准号: 7531812
• 负责人: PETER LESLIE DUTTON
• 金额: $ 35.93万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 1989
• 资助国家: 美国
• 起止时间: 1989-02-01 至 2010-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7531812
• 关键词: Age; Bioenergetics; Biological; Biological Models; Biomimetics; Catalysis; Catalytic Domain; Charge; Chemicals; Chemistry; Complex; Consensus; Coupled; Coupling; Cytochromes; Databases; Development; Devices; Diffusion; Disease; Electrodes; Electron Transport; Electron Transport Complex III; Electrons; Elements; Energy Supply; Engineering; Environment; Flavins; Food; Foundations; Freedom; Generations; Globin; Grant; Guidelines; Health; Heme; Human; Hydrocarbons; Hydrogen Peroxide; Hydroquinones; Integral Membrane Protein; Investigation; Kinetics; Left; Life; Ligand Binding; Light; Membrane; Membrane Potentials; Metabolic; Metabolism; Methods; Mitochondria; Modeling; Modification; Mono-S; NADH dehydrogenase (ubiquinone); Nature; Neurofibrillary Tangles; Niacinamide; Onset of illness; Operative Surgical Procedures; Oxidation-Reduction; Oxidoreductase; Oxygen; Oxygenases; Peroxides; Photosynthetic Reaction Centers; Physiological; Plague; Plant Roots; Plants; Positioning Attribute; Process; Property; Protein Engineering; Proteins; Protons; Quinones; Reaction; Reactive Oxygen Species; Regulation; Research Personnel; Research Project Grants; Resolution; Resources; Respiration; Respiratory Chain; Series; Site; Solutions; Stress; Structure; Superoxides; Testing; Thermodynamics; Time; Water; Work; base; cell growth regulation; cofactor; cytochrome c; cytochrome c oxidase; design; engineering design; hydroquinone; insight; microorganism; model design; nanoscale; novel; photosystem; respiratory; scaffold; simulation; theories

The energetic foundation of cellular activity and its regulation relies on a string of oxidoreductase proteins in biological oxidative and reductive metabolism and respiratory membrane energy conversion. The mechanisms of many catalytic sites of substrate oxidation-reduction and energy conversion, particularly in mitochondria! respiration, have proven to be difficult to access experimentally due to natural complexity and fragility. They remain poorly understood. Our proposal aims to reveal the natural engineering of the redox- coupled proton exchange and transfer operating at these sites of multi-electron cofactor and substrate oxidation-reduction. Our approach builds on our engineering guidelines for protein electron tunneling, strengthened in the last grant period, to inform the de novo design and assembly of simple and robust alpha- helical proteins intended to serve as protein-based models, maquettes, for multi-electron catalysis. Maquettes will be designed to provide the simplest water-soluble or trans-membrane structures that can capture the functional properties of natural redox centers. Their simplicity and adaptability allow us to investigate catalytic functional problems that remain unsolved in the respiratory chain. Maquettes will be activated with light and electrometric methods in solution and on electrodes to dissect step-by-step the thermodynamics and kinetics of electron transfer and proton exchange. Maquettes will incorporate all key two-electron, multi-proton cofactors/substrates quinone, nicotinamide and flavin, and the two-and four- electron substrate O2. We are positioned to focus on the problem of reversible energy conversion catalysis of hydroquinone-quinone in the Qo site of the cytochrome bd aiming to determine the mechanistic root of medically harmful short-circuits and radical generation. We will extend our maquette creation of a stable O2 ferrous heme state, to examine two- and four-electron O2 reduction and the physiologically important two- electron chemistries of NO and H2O2. With stable and adaptable maquettes, we can exploit physiological chemistry in the development of nanoscale devices. The breakdown of food by oxygen respiration in humans produces all the energy needed for a healthy life and is a central part of cellular regulation. It is normal that there is a steady slow release of oxygen radicals that over time cause cellular damage, aging and disease. Under stress, and in during many surgical procedures bursts of radicals can accelerate these deleterious processes. The research of this grant describes a new way to understand the processes underlying healthy oxidative energy supply and control as well as deleterious radical generation. With progress we will be better predict and track the onset of disease and act to slow it or reverse it.

细胞活性及其调节的能量基础依赖于一串氧化还原酶蛋白 生物氧化和还原的代谢和呼吸膜能量转化。这 底物氧化还原和能量转化的许多催化位点的机制，特别是在 线粒体！呼吸，由于自然复杂性和 脆弱性。他们仍然很了解。我们的建议旨在揭示氧化还原的自然工程 耦合质子交换和转移在多电子辅因子和底物的这些位置运行 还原氧化。我们的方法基于我们的蛋白质电子隧道工程指南， 在最后一个赠款期间得到加强，以告知从头设计和组装简单，强大的alpha- 螺旋蛋白旨在用作基于蛋白质的模型，用于多电子催化。 小块将设计为提供最简单的水溶性或跨膜结构 捕获天然氧化还原中心的功能特性。它们的简单性和适应性使我们能够 研究呼吸链中仍未解决的催化功能问题。马奎特会 用溶液和电极上的光和电量表激活，以分步剖析 电子传递和质子交换的热力学和动力学。 maquettes将包含所有钥匙 两电子，多质子辅助因子/底物奎因酮，烟酰胺和黄素，以及两个和四个 - 电子底物O2。我们可以专注于可逆的能量转化催化的问题 在细胞色素BD的QO位点中的氢醌 - 醌旨在确定机械根 医学上有害的短路和激进的一代。我们将扩展我们的稳定O2的制作 亚美蛋白血红素状态，以检查二元和四电子O2的降低以及生理上重要的两种 NO和H2O2的电子化学。凭借稳定且适应能力的毛孔，我们可以利用生理 纳米级设备开发的化学。 人类氧气呼吸对食物的细分产生健康生活所需的所有能量 是细胞调节的中心部分。正常的是，氧自由基的稳定缓慢释放 随着时间的流逝，会导致细胞损伤，衰老和疾病。在压力下，在许多手术期间 过程爆发可以加速这些有害过程。这笔赠款的研究 描述了一种理解健康氧化能量供应和控制过程的新方法 以及有害激进的一代。随着进步，我们将更好地预测并跟踪疾病的发作 并采取行动以减慢或逆转它。