Molecular analysis of the dimeric F1/F0-ATP synthase

二聚体 F1/F0-ATP 合酶的分子分析

• 批准号: 6693371
• 负责人: ROSEMARY A STUART
• 金额: $ 19.01万
• 依托单位: MARQUETTE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-01-01 至 2006-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6693371
• 关键词: binding sites; chromatography; crosslink; cytochrome oxidase; dimer; enzyme inhibitors; fungal genetics; fungal proteins; hydrogen transporting ATP synthase; intermolecular interaction; model design /development; molecular assembly /self assembly; physical model; protein purification; protein structure function; stoichiometry; western blottings

DESCRIPTION: (provided by applicant) We have recently shown that the yeast F1F0-ATP synthase complex exists as a dimer in the mitochondrial inner membrane. Isolation of the dimeric AlP synthase complex led to the identification of three new subunits of the complex. These subunits, Su e, Su g and Su k were found to be specifically associated with the dimeric form of the ATP synthase complex. The present proposal focuses on understanding the formation and function of the ATP synthase dimer. Su e appears to play a central role in the formation of the dimeric ATP synthase. Understanding both the function and the arrangement of Su e, and the other dimer specific-subunits, within the F0-sector represents a major goal of this research proposal. Second, we wish to analyze if the dimeric form of the F1F0-ATPase complex plays a central role in its ability to be inhibited by the natural inhibitor protein, IF1. It has been recently demonstrated that the natural inhibitor protein of the ATPase, IF1 (termed INH 1 in yeast) forms dimers and binds two neighboring F1-domains, concomitantly. We shall analyze whether formation of the dimeric F1F0-ATP synthase, mediated by Su e and Su g, plays a role in maintaining neighboring F1-sectors in close proximity to each other, so as to enable binding of the inhibitor protein dimer. Finally, our preliminary data suggests that the formation of the ATP synthase dimer is important for the maximal activity of the cytochrome oxidase complex. The analysis of the role of the dimeric ATP synthase and possible involvement with the cytochrome oxidase complex is also proposed. Since the assembly and oligomeric state of both of these complexes is anticipated to be similar in all eukaryotes, information gained from these studies should be helpful in the future analysis of human disorders stemming from enzymatic deficiencies in either the F1F0-ATP synthase or cytochrome oxidase complexes.

描述：（由申请人提供）我们最近表明酵母 F1F0-ATP合酶复合物以二聚体形式存在于线粒体内部 膜。二聚体 AlP 合酶复合物的分离导致 鉴定出该复合物的三个新亚基。这些亚基，Su e，Su g 和 Su k 被发现与二聚体形式特别相关 ATP合酶复合物。本提案的重点是了解 ATP合酶二聚体的形成和功能。 Su e 似乎演奏了 在二聚体 ATP 合酶的形成中发挥核心作用。了解两者 Su e 和另一个二聚体的功能和排列 F0 部门内的特定子单位代表了这一目标的主要目标 研究提案。其次，我们希望分析二聚体形式是否 F1F0-ATP酶复合物在其被抑制的能力中起着核心作用 天然抑制蛋白，IF1。最近已经证明， ATP 酶的天然抑制蛋白 IF1（在酵母中称为 INH 1）形成 二聚体并同时结合两个相邻的 F1 结构域。我们来分析一下 是否形成由 Su e 和 Su g 介导的二聚体 F1F0-ATP 合酶， 在保持相邻 F1 扇区彼此靠近方面发挥着作用 其他，以便能够结合抑制剂蛋白二聚体。最后，我们的 初步数据表明 ATP 合成酶二聚体的形成是 对于细胞色素氧化酶复合物的最大活性很重要。这 分析二聚体 ATP 合酶的作用以及可能的参与 还提出了细胞色素氧化酶复合物。自大会召开以来 预计这两种复合物的寡聚状态在所有方面都是相似的 真核生物，从这些研究中获得的信息应该有助于 对因酶缺乏引起的人类疾病的未来分析 F1F0-ATP 合酶或细胞色素氧化酶复合物。