Structure Function of the ATP Synthase

项目描述（由申请人提供）：该项目的长期目标是了解ATP合酶的结构/功能关系，因为它涉及ATP合酶的机制。最近，在一项合作研究中，我们能够获得酵母F1 atp酶的3.2-3.3 a图谱。这一重大进展现在使我们能够使用酵母遗传学和x射线晶体学的完整工具来提问和回答问题。本研究部分探讨了线粒体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。在本研究的第一个目标中，晶体结构分析将完成并改进到更高的分辨率。第二个目标将使用位点定向和随机诱变来研究γ -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F1F0 ATP合成酶活性位点形成中的作用。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合酶机制所需的最重要的剩余目标。这些实验将提供对ATP合酶结构功能的进一步了解，但也将提供ATP合酶亚基突变的功能后果的重要信息。最终，该项目将提供ATP合酶调控的关键信息，以及该调控对细胞在正常和疾病状态下提供组织和生物体所需的足够能量的整体能力的重要性。

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.