Structure and Function of Mammalian Copper Transporters

哺乳动物铜转运蛋白的结构和功能

• 批准号: 9353441
• 负责人: VINZENZ UNGER
• 金额: $ 33.09万
• 依托单位: NORTHWESTERN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-15 至 2019-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9353441
• 关键词: ATP phosphohydrolase; Address; Aerobic; Alzheimer' s Disease; Architecture; Behavior; Binding; Biological Assay; Biology; Carrier Proteins; Cell Compartmentation; Cell physiology; Cells; Cellular Membrane; Cellular biology; Cessation of life; Chemicals; Chemistry; Circadian Rhythms; Clinical; Complex; Copper; Creutzfeldt-Jakob Syndrome; Cryoelectron Microscopy; Crystallization; Cuprozinc Superoxide Dismutase; Dimensions; Dimerization; Disease; Drug Metabolic Detoxication; Electron Microscopy; Excretory function; Formulation; Goals; Hepatolenticular Degeneration; Homeostasis; Human; In Vitro; International; Ions; Length; Life; Lipid Bilayers; Liver; Location; Mammary gland; Maps; Mediating; Membrane; Menkes Kinky Hair Syndrome; Metabolism; Metals; Modeling; Molecular; Molecular Chaperones; Movement; Mutation; N-terminal; Negative Staining; Nerve Degeneration; Neuraxis; Organism; Oxidation-Reduction; Parkinson Disease; Pathway interactions; Pharmacotherapy; Phosphorylation; Phosphotransferases; Platinum; Process; Property; Proteins; Pump; Reactive Oxygen Species; Research; Resistance; Respiration; Role; Structure; Superoxide Dismutase; System; Ursidae Family; Variant; Wilson disease protein; Work; assay development; base; cancer therapy; conformational conversion; copper transporter 1; copper-transporting ATPase; dimer; electron crystallography; extracellular; human disease; in vitro Assay; in vivo; insight; metal metabolism; mutant; nanobodies; nervous system disorder; novel; orexin A receptor; particle; protein transport; reconstitution; reconstruction; scaffold; solute; structural biology; trafficking; tumor; uptake

SUMMARY At a chemical level, life is inconceivable without metals. This is reflected in the fact that imbalances in metal homeostasis invariably cause disease. Yet, metals are understudied, and their actions are largely taken for granted, in part because the complexity, and systemic integration of metal metabolism and strict cellular reliance on metals pose significant challenges to the exploration of these important contributors to cellular function. Addressing this shortcoming, our research aims to understand the molecular mechanisms that govern the cellular acquisition, distribution and excretion of one particular metal: copper. Although present in only small amounts, the coordination and redox chemistry of copper ions have become indispensible for cells because copper ions enable respiration and detoxification of reactive oxygen species – two fundamental processes that no aerobically growing organism can live without. Despite much progress in recent years, many fundamental question related to copper metabolism remain unanswered. Through work proposed in this application, we will answer the perplexing question how cells can beat the impossible odds of delivering copper to its final targets, we will develop an in vitro system that reconstitutes cellular copper uptake, and we will make major advances towards determining a structure of the copper pumping Wilson ATPase, ATP7B, that despite intense international efforts has remained an elusive target for structural studies. Aim 1, will be focused on the structure and function of the human copper importer hCTR1. Extending our previous work, we will determine the structure of hCTR1 in complex with CCS, the copper chaperone for Cu,Zn- superoxide dismutase 1, and develop an in vitro assay that will allow us to study mechanistic aspects of copper transport under controlled conditions. Aim 2, will focus on visualizing structure function correlates of the copper pump ATP7B mutations of which are the causative agent in Wilson Disease. Our studies will fill critical gaps in understanding of copper transport across cellular membranes and will advance a new paradigm posing that membrane scaffolding is essential for efficient intracellular copper distribution. Moreover, the anticipated results will make important contributions to understanding the molecular mechanisms underlying disorders that are associated with aberrant copper metabolism such as Wilson's disease, neurodegenerative conditions such as Parkinsons, Alzheimer's and Creutzfeldt- Jakob Disease, and tumor-resistance to platinum-based chemotherapeutics.

概括 在化学水平上，没有金属的寿命是不可想象的。这反映在以下事实中 金属稳态总是引起疾病。然而，金属被理解，它们的行为在很大程度上是 认为是理所当然的，部分是因为金属代谢的复杂性和全身整合 严格的蜂窝浮雕金属对这些重要的探索构成了重大挑战 细胞功能的贡献者。解决这一缺点，我们的研究旨在了解 控制一种特定的细胞获取，分布和排泄的分子机制 金属：铜。尽管仅出现少量，但铜的配位化学和氧化还原化学 由于铜离子能够呼吸和解毒，因此离子变得不可或缺 活性氧种 - 两个没有有氧生长有机体可以生存的两个基本过程 没有。 尽管近年来取得了很多进展，但许多基本问题与铜代谢有关 保持没有答复。通过本申请中提出的工作，我们将回答一个令人困惑的问题 细胞如何打败将铜输送到其最终目标的不可能的几率，我们将开发一个 重构蜂窝铜吸收的体外系统，我们将在 确定铜泵的结构Wilson ATPase，ATP7B，渴望强烈 国际努力仍然是结构研究的难以捉摸的目标。 AIM 1将重点放在 人类铜进口商HCTR1的结构和功能。扩展我们以前的工作，我们将 确定与CC的复合物中HCTR1的结构，CC，Cu，Zn-超氧化物的铜伴侣伴侣 验证酶1，并进行体外评估，这将使我们能够研究铜的机械方面 在受控条件下运输。 AIM 2，将专注于可视化结构功能的相关性 铜泵ATP7B突变是威尔逊病的病因。 我们的研究将填补了解跨细胞膜铜运输的关键空白，并将 推进新的范式，该范式是膜脚手架对于有效的细胞内铜至关重要 分配。此外，预期的结果将为理解 与异常铜代谢相关的疾病的分子机制 作为威尔逊氏病，神经退行性疾病，例如帕金森氏症，阿尔茨海默氏症和克鲁兹菲尔特 - 雅各布疾病，对基于铂的化学治疗药的肿瘤抗性。