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.

总结 在化学水平上，没有金属，生命是不可想象的。这反映在以下事实上： 体内金属平衡总是导致疾病。然而，金属是研究不足，他们的行动在很大程度上是 被认为是理所当然的，部分原因是金属代谢的复杂性和系统整合， 严格的细胞对金属的依赖对这些重要的 对细胞功能的贡献。针对这一缺陷，我们的研究旨在了解 控制细胞获取、分配和排泄一种特定的 金属：铜。尽管铜的存在量很小，但铜的配位和氧化还原化学 铜离子已经成为细胞不可或缺的，因为铜离子使呼吸和解毒， 活性氧-两个基本的过程，没有有氧生长的有机体可以生活 没有. 尽管近年来研究取得了很大进展，但许多与铜代谢相关的基础性问题仍然存在， 仍然没有答案。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 我们将开发一种方法来研究电池如何克服将铜输送到最终目标的不可能的困难 体外系统，重建细胞铜摄取，我们将取得重大进展， 确定铜泵威尔逊ATP酶，ATP 7 B的结构，尽管强烈 国际努力仍然是结构研究的一个难以捉摸的目标。目标1，将重点放在 人类铜输入者hCTR 1的结构和功能。为了扩展我们以前的工作，我们将 确定与CCS复合的hCTR 1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，使我们能够研究铜的机制方面 在受控条件下运输。目标2，将集中在可视化的结构功能相关的 铜泵ATP 7 B突变是威尔逊病的病原体。 我们的研究将填补理解铜跨细胞膜转运的关键空白， 提出了一种新的范式，提出膜支架对于有效的细胞内铜是必不可少的 分布此外，预期的结果将为理解 与异常铜代谢相关的潜在疾病的分子机制， 如威尔逊氏病，神经退行性疾病，如帕金森氏病，阿尔茨海默氏病和克罗伊茨费尔特- 雅各布病和肿瘤对铂类化疗药物的耐药性。