Structural and functional characterization of ATP synthase c-subunit leak channel and its role in AD pathogenesis

ATP合酶c亚基泄漏通道的结构和功能特征及其在AD发病机制中的作用

• 批准号: 10214094
• 负责人: Nelli Mnatsakanyan
• 金额: $ 15.81万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-05-15 至 2022-01-01
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10214094
• 关键词: Affinity; Alzheimer' s Disease; Alzheimer' s disease brain; Alzheimer' s disease model; Alzheimer' s disease pathology; Amyloid beta-Protein; Architecture; Binding; Biology; Brain; Calcium; Cell Death; Cell Death Induction; Cell Survival; Cells; Cleaved cell; Complex; Cryo-electron tomography; Cryoelectron Microscopy; Detergents; Development; Energy Metabolism; Environment; Event; Functional disorder; Glutamates; Goals; Hippocampus (Brain); Human; Immobilization; In Situ; Individual; Inner mitochondrial membrane; Interferometry; Kinetics; Lipids; Liposomes; Literature; Measures; Membrane; Mitochondria; Mitochondrial Proton-Translocating ATPases; Modeling; Molecular; Molecular Conformation; Mus; N-terminal; Neurodegenerative Disorders; Neurons; Oligomycins; Outer Mitochondrial Membrane; Pathogenesis; Pathologic; Pathway interactions; Permeability; Pharmacology; Play; Prevention strategy; Property; Proteins; Reporting; Research; Resolution; Role; Rupture; Signal Transduction; Structure; Synapses; System; Testing; Toxic effect; Transgenic Mice; cyclophilin D; cytochrome c; driving force; inhibitor/antagonist; mitochondrial dysfunction; mitochondrial membrane; mitochondrial permeability transition pore; monomer; mouse model; nanodisk; novel; novel therapeutics; overexpression; particle; patch clamp; prevent; reconstitution; sensor; time use

Abstract Mitochondrial and synaptic dysfunction are early pathological features and a driving force of Alzheimer's disease (AD) pathology. Aβ is found to accumulate abnormally in the brains of AD individuals and in an AD mouse models leading to mitochondrial Ca2+ overload and activation of mitochondrial permeability transition pore (mPTP). Prolonged opening of mPTP triggers outer mitochondrial membrane rupture, release of cytochrome c and activation of downstream cell death pathways. The mPTP has been at the center of extensive scientific research for the last several decades but it still remains as one of the most mysterious phenomena in biology today due to its controversial molecular composition and the lack of structural information of its pore. We have recently demonstrated the novel role of ATP synthase c-subunit ring in forming the channel of mPTP. Nevertheless, the gating mechanism of the ATP synthase c-subunit leak channel and conformational changes initiating its opening are yet to be discovered. In this proposal, we will use single-particle cryo-electron microscopy (cryo-EM) to identify the high- resolution structure and the open conformation of ATP synthase leak channel in the presence of channel modulators. We will also perform in situ structural analysis of ATP synthase in its functional environment within the Aβ-exposed primary hippocampal neurons and in mitochondria isolated from the mouse models of AD by using cryo-electron tomography (cryo-ET). In this project we will also investigate the direct role of ATP synthase leak channel as a novel cell death pathway in AD pathogenesis; we will test whether the pharmacological inhibition of this channel will rescue neurons from Aβ-induced cell death. Successful completion of this proposal will reveal the molecular mechanism(s) of mitochondrial permeability transition, the atomic structure of ATP synthase leak channel, and will aid in the development of new treatments for AD, targeting ATP synthase.

抽象的 线粒体和突触功能障碍是早期病理特征，也是疾病发生的驱动力。 阿尔茨海默病（AD）病理学。 AD 患者大脑中 Aβ 异常积累 个体和 AD 小鼠模型中导致线粒体 Ca2+ 过载和激活 线粒体通透性转换孔（mPTP）。 mPTP 的长时间打开会触发外部 线粒体膜破裂，细胞色素c释放并激活下游细胞 死亡途径。 mPTP 过去一直是广泛科学研究的中心 几十年来，它仍然是当今生物学中最神秘的现象之一 由于其分子组成存在争议且缺乏其孔的结构信息。 我们最近证明了 ATP 合酶 c 亚基环在形成 mPTP 通道。然而，ATP 合酶 c 亚基泄漏的门控机制 通道和启动其开放的构象变化尚未被发现。 在本提案中，我们将使用单粒子冷冻电子显微镜（cryo-EM）来识别高 存在下 ATP 合成酶泄漏通道的解析结构和开放构象 通道调制器。我们还将对其 ATP 合成酶进行原位结构分析。 暴露于 Aβ 的初级海马神经元和线粒体内的功能环境 通过使用冷冻电子断层扫描 (cryo-ET) 从 AD 小鼠模型中分离出来。在这个项目中 我们还将研究 ATP 合成酶泄漏通道作为新型细胞死亡途径的直接作用 AD发病机制；我们将测试该通道的药理抑制是否能挽救 Aβ 诱导的细胞死亡的神经元。该提案的成功完成将揭示 线粒体通透性转变的分子机制，ATP 的原子结构 合成酶泄漏通道，并将有助于开发针对 ATP 的 AD 新疗法 合酶。