Structure Function of the ATP Synthase

描述(申请人提供)：本项目的长期目标是了解ATP合成酶的结构/功能关系，因为它与ATP合成酶的机制有关。最近，在一项合作研究中，我们能够获得酵母F1 ATPase的3.2-3.3A图谱。这一重大进步现在允许我们使用酵母遗传学和X射线结晶学的全部工具来提问和回答问题。这项建议中的研究部分地调查了编码线粒体ATP合成酶亚单位的基因突变对ATP合成酶结构和功能的分子影响。这项研究的首要目标是完成晶体结构分析，并将其提高到更高的分辨率。第二个目标将使用定点突变和随机突变来研究伽马亚基相互作用的结构/功能关系。第三个目的是研究核苷酸结合在酶F1F0 ATP合成酶活性部位形成中的作用。这一最终目标代表了四个目标中最雄心勃勃的目标，但也代表了理解线粒体ATP合成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合成酶结构功能的进一步了解，也将提供关于ATP合成酶亚单位突变的功能后果的重要信息。最终，这个项目将提供关于ATP合成酶的调节以及这种调节对细胞的整体能力的重要性的关键信息，以提供组织和有机体在正常和疾病状态下所需的足够的能量。

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.