Molecular Mechanisms of Copper Transport

铜传输的分子机制

• 批准号: 9789970
• 负责人: Peng Yuan
• 金额: $ 34.34万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-30 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9789970
• 关键词: Address; Alzheimer' s Disease; Binding; Binding Sites; Bioavailable; Biochemical; Biochemical Process; Biochemistry; Biogenesis; Biological Assay; Biological Process; Biology; Biophysics; Cardiac; Cardiovascular Diseases; Carrier Proteins; Cells; Cellular Membrane; Coenzymes; Complement; Copper; Crystallization; Development; Diet; Disease; Drug Metabolic Detoxication; Enzymes; Essential Amino Acids; Eukaryota; Family; Foundations; Gene Mutation; Generations; Goals; Growth and Development function; Health; Heart Diseases; Hepatic; Hepatolenticular Degeneration; Homeostasis; Hormones; Human; Immune System Diseases; In Vitro; Integral Membrane Protein; Ions; Iron; Iron deficiency anemia; Knowledge; Link; Malignant Neoplasms; Membrane; Menkes Kinky Hair Syndrome; Metabolism; Methods; Mitochondria; Molecular; Molecular Chaperones; Mutagenesis; Nature; Nerve Degeneration; Neurodegenerative Disorders; Neurologic; Neurons; Neuropeptides; Organelles; Parkinson Disease; Pathologic; Pathology; Patients; Peripheral; Pharmacotherapy; Physiological; Physiology; Plasma; Positioning Attribute; Proteins; Regulation; Resolution; Respiration; Roentgen Rays; Role; Site-Directed Mutagenesis; Structure; Superoxides; Testing; Therapeutic; Toxic effect; Trace Elements; Work; X-Ray Crystallography; Yeasts; Zinc; base; biophysical properties; cofactor; human disease; in vitro Assay; in vivo; innovation; insight; large scale production; molecular modeling; mutant; novel therapeutics; reconstitution; three dimensional structure; uptake

!! ABSTRACT Molecular Mechanisms of Copper Transport Copper (Cu) is an essential trace element for growth and development because Cu acts as an indispensable cofactor for a variety of enzymes that are involved in multiple biological processes, and mutations of genes involved in Cu transport result in severe, even lethal, neurodegenerative diseases such as Wilson's disease and Menkes! syndrome. Abnormal Cu levels have also been linked to a range of pathological conditions, including Alzheimer's, Parkinson's, cardiovascular disease and cancer. Despite the overwhelming importance of Cu in health and disease, we have only rudimentary understanding of the molecular basis of Cu transport, with a lack of high-resolution three-dimensional structures and relevant biochemical and biophysical characterization that are essential for the development of appropriate mechanism-based therapeutics. Therefore, our long-term goal is to attain a comprehensive understanding of molecular mechanisms of Cu homeostasis using a combination of biochemical, biophysical, and structural approaches. In cells, appropriate Cu levels are tightly regulated by a sophisticated network of Cu-handling proteins, including Cu transporters, chaperones, and acceptors, to control the acquisition, distribution, and delivery of bioavailable Cu. Ubiquitous in eukaryotes, the copper transporter (Ctr) family of integral membrane proteins, Ctr1 and Ctr2, is involved in Cu transport across cellular membranes including both the plasma and intracellular organelle membranes. We have recently developed innovative methods for large-scale production of Ctr1 and Ctr2 transporter proteins, generation of diffraction-quality crystals, and for successful in vitro reconstitution assays. With these exciting preliminary developments, we are now able to combine X-ray crystallography, in vitro biochemical reconstitution, in vivo functional complementation assays, and site-directed mutagenesis to address Cu transport mechanisms. Specifically, we aim to determine the molecular basis of selectivity and permeation in Cu uptake by Ctr1, the underlying mechanism of zinc regulation in Ctr1, and molecular determinants for Cu transport in Ctr proteins. Our proposed work will provide atomic structures of Ctr1 and Ctr2 transporters in multiple functional states and uncover structural and molecular mechanisms of Cu transport. Detailed understanding of the mechanism, function, and regulation of Ctr proteins will open new therapeutic avenues for the treatment of a broad spectrum of diseases associated with disturbed Cu metabolism.

！！ 摘要 铜转运的分子机制 铜是生长发育所必需的微量元素，因为铜是人体不可缺少的 参与多种生物过程的各种酶的辅因子，以及基因突变 参与铜转运会导致严重的、甚至是致命的神经退行性疾病，如威尔逊病 还有门克斯！综合症。铜水平异常也与一系列病理状况有关， 包括阿尔茨海默氏症、帕金森氏症、心血管疾病和癌症。尽管具有压倒性的重要性 关于铜在健康和疾病中的作用，我们对铜运输的分子基础只有初步的了解， 缺乏高分辨率的三维结构和相关的生化和生物物理 对开发适当的以机制为基础的疗法至关重要的特征。 因此，我们的长期目标是全面了解铜的分子机制 使用生化、生物物理和结构方法相结合的动态平衡。在单元格中，适当 铜水平受到一个复杂的铜处理蛋白网络的严格调控，包括铜转运蛋白， 伴侣和受体，以控制生物有效铜的获取、分配和传递。无处不在 在真核生物中，铜转运蛋白(Ctr)家族的整合膜蛋白ctr1和ctr2参与了 铜的跨细胞膜运输包括胞质膜和胞内细胞器膜。我们 最近开发了大规模生产CTR1和Ctr2转运蛋白的创新方法， 产生衍射级晶体，并用于成功的体外重建试验。有了这些令人兴奋的东西 初步的发展，我们现在能够结合X射线结晶学，体外生化 铜的重组、体内功能互补分析和定点突变 传输机制。具体地说，我们的目标是确定选择性和渗透性的分子基础 CTR1对铜的吸收、CTR1中锌调控的潜在机制以及铜的分子决定因素 在CTR蛋白中的运输。我们拟议的工作将提供CTR1和Ctr2转运蛋白的原子结构 多种功能状态，揭示铜转运的结构和分子机制。详细 了解CTR蛋白的机制、功能和调节将开辟新的治疗途径 治疗与铜代谢紊乱相关的一系列疾病。