SOLID STATE NMR STUDIES OF F1F0 ATP SYNTHASE

F1F0 ATP 合酶的固态核磁共振研究

• 批准号: 6315526
• 负责人: Chad M Rienstra
• 金额: $ 3.24万
• 依托单位: COLUMBIA UNIV NEW YORK MORNINGSIDE
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-09-17 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6315526
• 关键词: bioenergetics; bioimaging /biomedical imaging; conformation; enzyme structure; hydrogen transport; hydrogen transporting ATP synthase; ion transport; membrane model; molecular dynamics; nuclear magnetic resonance spectroscopy; oxidative phosphorylation; structural biology

The synthesis and hydrolysis of ATP by the F1F0 ATP synthase are of central importance to all of biochemistry: the complex is the primary producer of biology's energetic currency. However, the details of how proton translocation couples with ATP synthesis are not well understood, because a high-resolution structure of the membrane-bound portion of the complex is not available. This assembled F0 portion, like many membrane protein complexes, has thus far resisted study by diffraction methods and solution NMR due to its heterogeneity and large effective molecular weight. Studies of ATP synthase (especially subunit c)--by incorporation of 13C, 15N, 19F, and 2H isotopic labels and solid state nuclear magnetic resonance (SSNMR) distance, torsion angle and relaxation measurements-will determine F0 structure and dynamics. In particular, the conformational changes that take place in subunit c upon uptake and release of protons at the essential aspartic acid residue will be carefully explored. These experiments will enhance the understanding of the molecular details of ion conduction, and the methodology developed and refined in the process will be directly applicable to many membrane proteins, responsible for energy transduction, cellular recognition, and many regulatory processes. The structural data and methods derived from this work, therefore, will be of utility for revealing the molecular mechanisms underlying membrane-related diseases such as cancer and AIDS.

F1F0 ATP 合酶对 ATP 的合成和水解对于所有生物化学都至关重要：该复合体是生物能量货币的主要生产者。 然而，质子易位如何与 ATP 合成耦合的细节尚不清楚，因为无法获得复合物膜结合部分的高分辨率结构。 这种组装的 F0 部分，像许多膜蛋白复合物一样，由于其异质性和大的有效分子量，迄今为止一直抵制衍射方法和溶液核磁共振的研究。对 ATP 合酶（尤其是 c 亚基）的研究——通过结合 13C、15N、19F 和 2H 同位素标记以及固态核磁共振 (SSNMR) 距离、扭转角和弛豫测量——将确定 F0 结构和动力学。 特别是，将仔细探索在必需的天冬氨酸残基处摄取和释放质子时亚基c中发生的构象变化。 这些实验将增强对离子传导分子细节的理解，在此过程中开发和完善的方法将直接适用于许多负责能量转导、细胞识别和许多调节过程的膜蛋白。 因此，从这项工作中获得的结构数据和方法将有助于揭示癌症和艾滋病等膜相关疾病的分子机制。