Structure and regulation 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。这在心脏组织中尤其重要，因为ATP合酶占有氧条件下产生的大部分ATP。此外，心脏ATP合酶的调节已被证明发生在缺血条件下。最近，在一项合作研究中，我们能够获得酵母F1 atp酶的3.0-3.3 a电子密度图。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。本研究的第一个目的是验证一个假设，即ATP合酶的组装可以在缺乏关键亚基的情况下发生，并且由此产生的复合物负责线粒体的解偶联。第二个目的是测试一类突变（mgi）的机制假说，这些突变是在ATP合酶的亚基中分离出来的。第三个目标将利用x射线晶体学获得含有mgi突变的f1 - atp酶的高分辨率结构。第四个也是最后一个目标将检验ATP合酶的天然抑制剂阻止γ -亚基单向旋转的假设。这些实验将提供对ATP合酶的结构和功能的进一步了解，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. This is particularly important in cardiac tissue since the ATP synthase accounts for majority of ATP made under aerobic conditions. Furthermore, regulation of the ATP synthase in heart has been shown to occur under ischemic conditions. Recently, in a collaborative study, we were able to obtain a 3.0-3.3 A electron density 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. The first aim of this study tests a hypothesis that assembly of ATP synthase can occur in the absence of key subunits and the resulting complex is responsible of the uncoupling of the mitochondria. The second aim tests the hypothesis for the mechanism of a class of mutations, mgi, isolated in subunits of the ATP synthase. The third aim will utilize x-ray crystallography to obtain a high-resolution structure of the F1-ATPase containing the mgi mutations. The fourth and final aim will test the hypothesis that the natural inhibitor of the ATP synthase prevents the unidirectional rotation of the gamma-subunit. These experiments will provide a further understanding on the structure and function of the ATP synthase, but will also provide important information on the functional consequences of mutations in subunits of the 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 states.