Molecular Mechanisms of Copper Transport

铜传输的分子机制

• 批准号: 10213151
• 负责人: Peng Yuan
• 金额: $ 34.45万
• 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/10213151
• 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; 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.

摘要 铜离子传输的分子机制 铜（Cu）是生长和发育所必需的微量元素，因为Cu充当不可或缺的 参与多种生物过程的多种酶的辅因子，以及基因突变 参与铜转运导致严重的、甚至致命的神经退行性疾病，如威尔逊病 和门克斯综合征异常的铜水平也与一系列病理状况有关， 包括老年痴呆症、帕金森症、心血管疾病和癌症。尽管有压倒性的重要性 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， 由于缺乏高分辨率的三维结构和相关的生物化学和生物物理 这些特征对于开发适当的基于机制的治疗剂至关重要。 因此，我们的长期目标是全面了解铜的分子机制 使用生物化学、生物物理学和结构方法的组合来维持体内平衡。在细胞中，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， 分子伴侣和受体，以控制生物可利用的Cu的获取、分布和递送。无处不在 在真核生物中，铜转运蛋白（Ctr）家族的完整膜蛋白Ctr 1和Ctr 2参与了 铜跨细胞膜转运，包括质膜和细胞内细胞器膜。我们 最近开发了用于大规模生产Ctr 1和Ctr 2转运蛋白的创新方法， 产生衍射质量的晶体，并用于成功的体外重构测定。有了这些令人兴奋的 初步的发展，我们现在能够结合联合收割机X射线晶体学，体外生化 重组，体内功能互补测定和定点诱变来解决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， Ctr 1的Cu吸收，Ctr 1中锌调节的潜在机制，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，并揭示铜运输的结构和分子机制。详细 对Ctr蛋白的机制、功能和调节的理解，将为癌症的治疗开辟新的途径。 治疗与铜代谢紊乱相关的广谱疾病。