Structural biology of copper homeostasis

铜稳态的结构生物学

• 批准号: 7560072
• 负责人: AMY C. ROSENZWEIG
• 金额: $ 30.14万
• 依托单位: NORTHWESTERN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-02-01 至 2012-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7560072
• 关键词: ATP phosphohydrolase; ATP7A protein; Adenylyl Imidodiphosphate; Antineoplastic Agents; Archaeoglobus fulgidus; Arts; Binding; Biological Models; Biological Process; C-terminal; Cocrystallography; Complex; Copper; Crystallization; DNA; DNA Binding; Data; Databases; Disease; Elements; Enzymes; Future; Gene Expression Regulation; Genetic Transcription; Helix-Turn-Helix Motifs; Hepatolenticular Degeneration; Hereditary Disease; Homeostasis; Human; Immunoglobulin Fragments; Ions; Lead; Ligands; Link; Mediating; Membrane; Membrane Proteins; Membrane Transport Proteins; Menkes Kinky Hair Syndrome; Metabolism; Metals; Microfluidics; Molecular; Molecular Chaperones; Molecular Conformation; Mutation; N Domain; N-terminal; Pathway interactions; Phase; Physiological; Play; Property; Protein Binding; Proteins; Reactive Oxygen Species; Recombinants; Research; Resistance; Role; Route; Screening procedure; Specificity; Structure; Sulfolobus solfataricus; Techniques; Tertiary Protein Structure; Transmembrane Domain; Variant; Work; base; cofactor; human disease; hyperthermophile; insight; metalloregulatory protein; novel; programs; protein complex; protein function; protein protein interaction; protein structure; structural biology; toxic metal; trafficking; two-dimensional

DESCRIPTION (provided by applicant): Copper serves as a cofactor for many enzymes involved in important biological processes, but can also facilitate the formation of toxic organic and oxygen radicals. A host of proteins, including membrane transporters, metallochaperones, and metalloregulatory proteins, maintains intracellular copper concentrations such that copper ions are provided to essential enzymes, but do not accumulate to deleterious levels. Understanding how these proteins function on the molecular level is the theme of this ongoing research program. Despite significant progress toward determining how soluble copper trafficking proteins bind metal ions, recognize physiological partners, and facilitate metal ion transfer, there are large gaps in the current understanding of copper homeostasis, particularly regarding copper translocation across membranes by P1B- type ATPases. Importantly, mutations in human Cu+transporting P1B-type ATPases lead to Wilson disease and Menkes syndrome, serious disorders of copper metabolism. To complete the molecular picture of copper trafficking and to advance understanding of Cu+ATPases, a model system from the hyperthermophile Archaeoglobus fulgidus is being investigated. The A. fulgidus CopA Cu+ATPase contains all of the structural elements that are present in the human Wilson and Menkes disease proteins, including soluble metal binding domains (MBDs), an ATP binding domain (ATPBD), and an actuator domain (A-domain). Other components of the A. fulgidus pathway include a novel CopZ copper chaperone and a putative transcriptional regulator, CopT. The proposed research involves biophysical and structural characterization of CopA, CopZ, and CopT. The soluble CopA MBDs will be structurally characterized and protein-protein interactions between the MBDs and the CopZ chaperone will be investigated. The copper binding properties, potential interactions with CopZ, and structure of CopT will be probed. Finally, state-of-the-art crystallization techniques for membrane proteins will be applied to CopA, and structures determined in multiple conformations. These data will provide molecular insight into the molecular basis for Wilson and Menkes diseases as well as adding to the database of membrane protein structures. A number of human diseases are linked to deficiencies in cellular handling of copper, which is an essential yet potentially toxic metal ion. This project will provide a molecular picture of the type of protein that is defective in Wilson disease and Menkes syndrome, both genetic disorders of copper metabolism. These same proteins may also be associated with resistance to anticancer drugs.

描述（由申请人提供）：铜是参与重要生物过程的许多酶的辅助因子，但也可以促进有毒有机自由基和氧自由基的形成。许多蛋白质，包括膜转运蛋白、金属伴侣和金属调节蛋白，维持细胞内铜浓度，从而将铜离子提供给必需酶，但不会累积到有害水平。了解这些蛋白质如何在分子水平上发挥作用是这个正在进行的研究项目的主题。尽管在确定可溶性铜运输蛋白如何结合金属离子、识别生理伙伴和促进金属离子转移方面取得了重大进展，但目前对铜稳态的理解仍存在很大差距，特别是在 P1B 型 ATP 酶跨膜铜易位方面。重要的是，人类 Cu+ 转运 P1B 型 ATP 酶的突变会导致威尔逊病和门克斯综合征，这是严重的铜代谢紊乱。为了完成铜运输的分子图谱并促进对 Cu+ATP 酶的理解，正在研究来自超嗜热古生球菌 fulgidus 的模型系统。 A. fulgidus CopA Cu+ATPase 包含人类威尔逊病和门克斯病蛋白中存在的所有结构元件，包括可溶性金属结合域 (MBD)、ATP 结合域 (ATPBD) 和致动域 (A-领域）。 A. fulgidus 途径的其他成分包括新型 CopZ 铜伴侣和假定的转录调节因子 CopT。拟议的研究涉及 CopA、CopZ 和 CopT 的生物物理和结构表征。将对可溶性 CopA MBD 进行结构表征，并研究 MBD 和 CopZ 分子伴侣之间的蛋白质-蛋白质相互作用。将探讨铜的结合特性、与 CopZ 的潜在相互作用以及 CopT 的结构。最后，最先进的膜蛋白结晶技术将应用于 CopA，并确定多种构象的结构。这些数据将为威尔逊病和门克斯病的分子基础提供分子洞察，并添加到膜蛋白结构的数据库中。 许多人类疾病与细胞处理铜的缺陷有关，铜是一种必需但具有潜在毒性的金属离子。该项目将提供威尔逊病和门克斯综合征中缺陷蛋白质类型的分子图谱，这两种疾病都是铜代谢的遗传性疾病。这些相同的蛋白质也可能与抗癌药物的耐药性有关。