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 铜运输的分子机制 铜（CU）是生长和发展的必不可少的痕量元素，因为CU充当必不可少的 用于多种生物学过程的多种酶的辅因子和基因突变 参与CU运输会导致严重，甚至致命的神经退行性疾病，例如威尔逊氏病 和Menkes综合征。异常CU水平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。尽管非常重要 CU在健康和疾病方面，我们只对Cu转运的分子基础有基本的了解， 缺乏高分辨率的三维结构以及相关的生化和生物物理 对于开发适当的基于机制的治疗剂至关重要的表征。 因此，我们的长期目标是获得对Cu分子机制的全面理解 使用生化，生物物理和结构方法结合使用稳态。在细胞中，适当 CU水平由复杂的Cu处理蛋白网络严格调节，包括Cu转运蛋白， 伴侣和受体，以控制可生物利用Cu的获取，分发和交付。无处不在 在真核生物中，整体膜蛋白CTR1和CTR2的铜转运蛋白（CTR）家族参与 CU跨细胞膜的转运，包括血浆和细胞内细胞器膜。我们 最近开发了用于大规模生产CTR1和CTR2转运蛋白的创新方法， 生成衍射质量的晶体，并为成功的体外重建测定法。这些令人兴奋的 初步发展，我们现在能够结合X射线晶体学，体外生化 重构，体内功能互补测定和定向诱变以解决Cu 运输机制。具体而言，我们旨在确定选择性和渗透的分子基础 CTR1的Cu摄取，CTR1的基本机理Ctr1中的基本机制和Cu的分子决定因素 CTR蛋白中的运输。我们提出的工作将在CTR1和CTR2转运蛋白的原子结构中提供 多个功能状态以及CU转运的结构和分子机制。详细的 了解CTR蛋白的机制，功能和调节将为新的治疗途径开放 与CU代谢障碍有关的广泛疾病的治疗。