Structure Function of the ATP Synthase

项目描述（由申请人提供）：本项目的长期目标是了解ATP合酶的结构/功能关系，因为它与ATP合酶的机制有关。最近，在一项合作研究中，我们能够获得3.2-3.3 A的酵母F1 ATP酶图谱。这一重大进展现在允许我们使用酵母遗传学和X射线晶体学的全部工具来提问和回答问题。在这项研究中，部分研究了线粒体ATP合酶亚基编码基因突变对ATP合酶结构和功能的分子影响。在这项研究的第一个目标，晶体结构分析将完成和改进，甚至进一步的分辨率。第二个目标将使用定点突变和随机突变来研究γ-亚基相互作用的结构/功能关系。第三个目标是研究核苷酸结合在形成酶F1 F0 ATP合成酶活性位点上的作用。这个最终目标代表了4个目标中最雄心勃勃的目标，但也代表了理解线粒体ATP合酶机制所需的唯一最重要的剩余目标。这些实验将提供对ATP合酶的结构功能的进一步理解，但也将提供重要的信息，在ATP合酶的亚基突变的功能后果。最终，该项目将提供有关ATP合酶调节的关键信息，以及这种调节对细胞整体能力的重要性，以提供正常和疾病状态下组织和生物体所需的足够能量。

DESCRIPTION (provided by applicant): The long-term goal of this project is to gain an understanding on the structure/function relationship of the ATP synthase as it relates to mechanism of the ATP synthase. Recently, in a collaborative study, we were able to obtain a 3.2-3.3 A map of the yeast F1 ATPase. This major advance now allows us to ask and answer questions using the full tools of yeast genetics and x-ray crystallography. The studies in this proposal, in part, investigate the molecular impact of mutations in genes encoding subunits of the mitochondrial ATP synthase on the structure and function of the ATP synthase. In first aim of this study, the crystal structure analysis will be completed and improved to even further resolution. The second aim will use both site directed and random mutagenesis to investigate the structure/function relationship of the interactions of the gamma-subunit. The third aim will investigate the role of nucleotide binding on forming the active sites of the enzyme, F1F0 ATP synthase. This final aim represents the most ambitious aim of the 4, but also represents the single most important remaining goal needed in the understanding of the mechanism of the mitochondrial ATP synthase. These experiments will provide a further understanding on the structure function of the ATP synthase, but will also provide important information on the functional consequences of mutations in subunits of he ATP synthase. Ultimately, this project will provide critical information on the regulation of the ATP synthase and the importance of this regulation on the overall capabilities of the cell to provide enough energy as demanded by tissue and organism in both the normal and disease state.