Structure and Function of the proton-turbine of the ATP-Synthase

ATP 合酶质子涡轮的结构和功能

• 批准号: 7299157
• 负责人: PETRA FROMME
• 金额: $ 24.02万
• 依托单位: ARIZONA STATE UNIVERSITY-TEMPE CAMPUS
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-01 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7299157
• 关键词: ATP Synthesis Pathway; Alzheimer' s Disease; Binding; Binding Sites; Chloroplasts; Complex; Coupling; Crystallization; Crystallography; Cyanobacterium; Defect; Degradation Pathway; Disease; Enzymes; Goals; Human; Individual; Ions; Lead; Life; Membrane; Membrane Proteins; Mitochondria; Mitochondrial Proton-Translocating ATPases; Molecular; Molecular Motors; Motor; Multienzyme Complexes; Nucleotides; Organism; Pathway interactions; Plants; Play; Preparation; Probability; Protein Overexpression; Protein Subunits; Proteins; Protons; Red Algae; Retinitis Pigmentosa; Roentgen Rays; Role; Source; Spielmeyer-Vogt Disease; Spinach - dietary; Structure; Syndrome; System; Trees; Work; X-Ray Crystallography; base; enzyme mechanism; nano; protein structure; response; sodium ion; stoichiometry

DESCRIPTION (provided by applicant): One of the most important proteins in energy transduction is the ATP-Synthase (FOF1). This membrane- bound large protein complex is present in nearly all organisms and is essential for all higher life on earth, including humans. Slight defects in function, assembly and the degradation pathways of the enzyme play important roles in severe diseases such as the Batten's disease, Alzheimer's disease and the Retinitis Pigmentosa Syndrome. Furthermore, human mitochondrial ATP Synthase is highly regulated in response to cellular energy needs. The aim of this project is to unravel the structure and catalytic mechanism of this enzyme. The ATP Synthase functions as a molecular (nano) motor, thereby catalyzing the synthesis of ATP from ADP and Pi driven by a transmembrane electrochemical potential of protons or sodium ions. The enzyme complex consists of two distinct structural and functional domains: A membrane intrinsic proton translocation system (the FO part), which is structurally connected by at least two "stalks" to the membrane extrinsic domain (the F1 part), which in turn harbors the nucleotide binding sites. While several structures of the membrane extrinsic F1 part and individual protein subunits of the ion conduction FO part have been determined by X-ray structure crystallography, the molecular elucidation of the coupling mechanism still suffers from the lack of information on the structure of the complete intact ATP Synthase and the membrane intrinsic proton translocation machinery. The goal of this project is to determine the structure of proton turbine of the ATP-Synthase and discover the mechanism of the coupling between proton transfer and ATP-synthesis. The project aims to crystallize the intact ATP Synthase, the proton conducting FO part as well as the c-ring rotor of the enzyme, which differs in its oligomeric state between different organisms. The crystals will be used to determine the structure of the protein complexes by X-ray crystallography. The structural information will form the basis for the discovery of the mechanism of the dynamic energy coupling in the catalytic cycle of the enzyme.

描述（由申请人提供）：能量转导中最重要的蛋白质之一是ATP合酶（FOF 1）。这种膜结合的大型蛋白质复合物存在于几乎所有的生物体中，并且是地球上所有高等生命（包括人类）所必需的。该酶的功能、组装和降解途径的轻微缺陷在严重疾病如Batten病、阿尔茨海默病和视网膜色素变性综合征中起重要作用。此外，人类线粒体ATP合酶响应细胞能量需求而高度调节。本项目的目的是揭示这种酶的结构和催化机制。ATP合酶作为分子（纳米）马达发挥作用，从而催化由质子或钠离子的跨膜电化学势驱动的ADP和Pi合成ATP。该酶复合物由两个不同的结构和功能结构域组成：膜固有质子转运系统（FO部分），其在结构上通过至少两个“茎”连接到膜外在结构域（F1部分），而膜外在结构域又包含核苷酸结合位点。虽然膜的外部F1部分和离子传导FO部分的单个蛋白亚基的几种结构已经通过X-射线结构晶体学确定，但对偶联机制的分子阐明仍然缺乏关于完整完整的ATP合酶和膜的内部质子转运机制的结构的信息。本项目的目标是确定ATP合成酶质子涡轮机的结构，揭示质子转移与ATP合成耦合的机理。该项目旨在结晶完整的ATP合酶，质子传导FO部分以及酶的C环转子，其在不同生物体之间的低聚状态不同。晶体将用于通过X射线晶体学确定蛋白质复合物的结构。这些结构信息将为发现酶催化循环中的动态能量耦合机制奠定基础。