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 在化学层面上，没有金属，生命是不可想象的，这反映在金属的不平衡这一事实上。 金属稳态总是会导致疾病，然而，人们对金属的研究还不够充分，而且它们的作用在很大程度上还不清楚。 被认为是理所当然的，部分原因是金属代谢和金属代谢的复杂性和系统性整合 细胞对金属的严格依赖对探索这些重要的金属提出了重大挑战 针对这一缺点，我们的研究旨在了解细胞功能的贡献者。 控制细胞获取、分布和排泄一种特定物质的分子机制 金属：铜，虽然含量很少，但铜的配位和氧化还原化学性质。 离子已成为细胞不可或缺的元素，因为铜离子能够促进细胞的呼吸和解毒。 活性氧——有氧生长的生物体无法生存的两个基本过程 没有。 尽管近年来取得了很大进展，但与铜代谢相关的许多基本问题 通过本申请中提出的工作，我们将回答这个令人困惑的问题。 细胞如何克服不可能的困难将铜输送到最终目标，我们将开发一种新的方法 重建细胞铜摄取的体外系统，我们将在以下方面取得重大进展 确定铜泵威尔逊 ATP 酶 (ATP7B) 的结构，尽管强度很大 国际仍然是结构研究的一个难以捉摸的目标，目标 1 将重点关注 人类铜输入蛋白 hCTR1 的结构和功能 扩展我们之前的工作，我们将 确定 hCTR1 与 CCS 复合物的结构，CCS 是 Cu,Zn-超氧化物的铜伴侣 歧化酶 1，并开发一种体外测定方法，使我们能够研究铜的机制方面 目标 2 将重点关注可视化结构功能相关性。 铜泵 ATP7B 突变是威尔逊病的致病因素。 我们的研究将填补理解铜跨细胞膜运输的关键空白，并将 提出了一种新的范式，认为膜支架对于有效的细胞内铜至关重要 此外，预期结果将为理解分布做出重要贡献。 与异常铜代谢相关的疾病的分子机制，例如 如威尔逊氏病、神经退行性疾病，如帕金森病、阿尔茨海默病和克伊茨费尔特病 - 雅各布病和肿瘤对铂类化疗药物的耐药性。