Copper Protein In Metal and Oxidant Stress Responses

金属中的铜蛋白和氧化应激反应

• 批准号: 7154133
• 负责人: THOMAS V O'HALLORAN
• 金额: $ 31.18万
• 依托单位: NORTHWESTERN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-02-01 至 2009-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7154133
• 关键词: Active Sites; Address; Amyotrophic Lateral Sclerosis; Antioxidants; Biochemical; Biochemical Genetics; Biological; Cells; Cellular biology; Chemicals; Chemistry; Complex; Condition; Copper; Cytoplasm; Data; Dioxygen; Disease; Disulfides; Docking; Ensure; Enzymes; Familial Amyotrophic Lateral Sclerosis; Family; Funding; Genetic; Goals; Grant; Homeostasis; Human; In Vitro; Ions; Kinetics; Menkes Kinky Hair Syndrome; Metalloproteins; Metals; Methods; Mitochondria; Modeling; Molecular; Molecular Chaperones; Mutation; Nature; Neurodegenerative Disorders; Numbers; Oxidants; Oxidative Stress; Oxygen; Pathology; Pathway interactions; Phase; Philosophy; Physiological; Physiology; Play; Post-Translational Regulation; Process; Proteins; Reaction; Reactive Oxygen Species; Reagent; Regulation; Research Support; Role; Signal Transduction; Site; Standards of Weights and Measures; Structure; Techniques; Temperature; Testing; Thermodynamics; Variant; Work; Yeasts; base; biological adaptation to stress; cofactor; copper zinc superoxide dismutase; crosslink; disulfide bond; gain of function; gain of function mutation; human disease; in vivo; insight; mutant; oxidation; protein aggregate; protein function; protein protein interaction; receptor; research study; tool; trafficking

DESCRIPTION (provided by applicant): The metallochaperone proteins Atx1 and CCS (copper chaperone for copper, zinc superoxide dismutase, SOD1) are soluble metal receptor proteins that function to guide and protect the metal ion while facilitating appropriate partnerships within the cell. The Atx1-like proteins ensure the facile delivery of a Cu(l) cofactor to intracellular targets in the secretory pathway while CCS has a more complex structure and function. New results suggest that this metallochaperone requires oxidants such as oxygen to complete formation of the mature and active state of SOD1. Elucidating the mechanisms of these processes will provide keys to understanding the cell biology of copper in pathological conditions, such as Wilson and Menkes disease and familial amyotrophic lateral sclerosis (fALS). Kinetic, thermodynamic and structure-function studies of the metallochaperones and their physiological targets will test the hypothesis that these proteins function by lowering the activation barrier for Cu-transfer to partner proteins but maintain high barriers for transfer to other sites. While the multidomain copper chaperone CCS neither detoxifies copper or reactive oxygen species (ROS), new results suggest that it plays a role in posttranslational regulation of oxidative stress responses: as oxidative stress increases, CCS facilitates the correct disulfide bond formation in its target, apoSODL These mechanistic, physiological and structural studies will provide the basis for a more complete understanding of metal trafficking and homeostasis in disease. The newly developed tools and reagents will be used to address roles of copper proteins in neurodegenerative diseases, as well as the emerging connections between copper cell biology and oxygen physiology. For instance these studies will test an emerging model for the gain of function mutations in SOD1 that cause fALS: the immature disulfide reduced forms of the disease causing proteins are completely unfolded at physiological temperature and readily become insoluble aggregates upon formation of inappropriate disulfide crosslinks.

描述(申请人提供)：金属配位体蛋白Atx1和CCS(铜，锌超氧化物歧化酶的铜伴侣，SOD1)是可溶的金属受体蛋白，其功能是引导和保护金属离子，同时促进细胞内适当的伙伴关系。类ATX1蛋白确保了铜(L)辅因子在细胞内的分泌途径，而CCS具有更复杂的结构和功能。新的结果表明，这种金属配位体需要氧气等氧化剂来完成SOD1的成熟和活性状态的形成。阐明这些过程的机制将为理解铜在诸如Wilson和Menkes病和家族性肌萎缩侧索硬化症(FALS)等病理条件下的细胞生物学提供关键。对金属配位配位蛋白及其生理靶标的动力学、热力学和结构功能的研究将验证这样的假设，即这些蛋白质通过降低铜向配对蛋白转移的激活障碍而保持向其他位置转移的高障碍来发挥作用。虽然多结构域铜伴侣CCS不能解毒铜或活性氧物种(ROS)，但新的结果表明，它在氧化应激反应的翻译后调节中发挥作用：随着氧化应激的增加，CCS促进其靶标apoSODL中正确的二硫键形成。这些机制、生理和结构研究将为更全面地了解疾病中的金属运输和动态平衡提供基础。新开发的工具和试剂将用于研究铜蛋白在神经退行性疾病中的作用，以及铜细胞生物学和氧气生理之间的新出现的联系。例如，这些研究将测试一种新的模型，用于获得导致FALS的SOD1功能突变：致病蛋白的未成熟二硫键还原形式在生理温度下完全展开，并在形成不适当的二硫键交联物时很容易成为不可溶的聚集体。