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）：铜是许多参与重要生物过程的酶的辅因子，但也可以促进有毒有机和氧自由基的形成。许多蛋白质，包括膜转运蛋白，金属伴侣和金属调节蛋白，都保持细胞内铜浓度，从而提供铜离子以提供必需酶，但不会积聚到有害水平。了解这些蛋白质在分子水平上的作用是该正在进行的研究计划的主题。尽管在确定可溶性铜运输蛋白如何结合金属离子，识别生理伙伴并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中存在很大的差距，尤其是关于P1B-类型ATPases跨膜的铜易位。重要的是，人类Cu+运输P1B型ATPases的突变导致Wilson病和Menkes综合征，这是铜代谢的严重疾病。为了完成铜运输的分子图并提高对Cu+Atpases的了解，正在研究来自高疗法Archaeoglobus fulgidus的模型系统。 Fulgidus copa cu+ATPase包含人类Wilson和Menkes疾病蛋白中存在的所有结构元素，包括可溶性金属结合结构域（MBD），ATP结合结构域（ATPBD）和执行器结构域（A-Domain）。 A. fulgidus途径的其他组件包括新型Copz铜伴侣和推定的转录调节器Copt。提出的研究涉及Copa，Copz和Copt的生物物理和结构表征。可溶性COPA MBD将在结构上表征，并研究MBD和COPZ伴侣之间的蛋白质 - 蛋白质相互作用。将探测铜结合特性，与COPZ的潜在相互作用以及COPT的结构。最后，膜蛋白的最新结晶技术将应用于Copa，并以多种构象确定的结构。这些数据将为Wilson和Menkes疾病的分子基础提供分子见解，并将其添加到膜蛋白结构的数据库中。 许多人类疾病与铜的细胞处理中的缺陷有关，这是一种必不可少但可能有毒的金属离子。该项目将提供一张分子图片，说明在威尔逊疾病和Menkes综合征中有缺陷的蛋白质类型，这是铜代谢的遗传疾病。这些相同的蛋白质也可能与对抗癌药物的抗性有关。