Molecular Mechanisms of Copper Transport

铜传输的分子机制

• 批准号: 10418771
• 负责人: Peng Yuan
• 金额: $ 6.54万
• 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 2023-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10418771
• 关键词: Address; Alzheimer' s Disease; Binding; Binding Sites; Biochemical; Biochemical Process; Biochemistry; Biogenesis; Biological Assay; Biological Process; Biology; Biophysics; Cardiac; Cardiovascular Diseases; Carrier Proteins; Cells; Cellular Membrane; Coenzymes; Complement; Copper; Crystallization; Development; 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; dietary; 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.

抽象的 铜传输的分子机制 铜 (Cu) 是生长发育必需的微量元素，因为 Cu 是生长发育中不可缺少的微量元素。 参与多种生物过程和基因突变的多种酶的辅因子 参与铜转运会导致严重甚至致命的神经退行性疾病，例如威尔逊氏病 和门克斯综合症。铜水平异常也与一系列病理状况有关， 包括阿尔茨海默病、帕金森病、心血管疾病和癌症。尽管具有压倒性的重要性 铜在健康和疾病中的作用，我们对铜转运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物化学和生物物理 对于开发适当的基于机制的疗法至关重要的表征。 因此，我们的长期目标是全面了解 Cu 的分子机制。 结合生物化学、生物物理和结构方法来实现体内平衡。在细胞内，适当 铜水平受到复杂的铜处理蛋白网络的严格调节，包括铜转运蛋白、 伴侣和受体，以控制生物可利用铜的获取、分配和传递。无处不在 在真核生物中，整合膜蛋白的铜转运蛋白 (Ctr) 家族 Ctr1 和 Ctr2 参与 铜跨细胞膜运输，包括质膜和细胞内细胞器膜。我们 最近开发了大规模生产 Ctr1 和 Ctr2 转运蛋白的创新方法， 生成衍射质量的晶体，并成功进行体外重构测定。有了这些激动人心的 经过初步开发，我们现在能够将 X 射线晶体学、体外生化结合起来 重组、体内功能互补测定和定点诱变以解决 Cu 运输机制。具体来说，我们的目标是确定选择性和渗透的分子基础 Ctr1 对 Cu 的吸收、Ctr1 中锌调节的基本机制以及 Cu 的分子决定因素 Ctr 蛋白的运输。我们提出的工作将提供 Ctr1 和 Ctr2 转运蛋白的原子结构 多种功能状态并揭示铜传输的结构和分子机制。详细的 了解 Ctr 蛋白的机制、功能和调节将为以下疾病开辟新的治疗途径 治疗与铜代谢紊乱相关的多种疾病。