Mechanisms of metal ion regulation in human cells

描述(由申请人提供)：该计划的总体目标是在正常和疾病的细胞和组织中产生一幅关于人类铜稳态的全面机制图。铜是胚胎发育、结缔组织形成、神经元髓鞘形成、自由基解毒和许多其他生理过程所必需的营养物质。缺铜对人类的生长发育有害，而过多的铜则有毒。铜稳态的失调与严重的代谢和神经紊乱有关，这可能会影响生命中任何阶段的患者。在这项计划项目中，控制人类细胞和组织中铜分布的机制将由一组经验丰富的研究人员组成，他们在结构生物学、膜运输、金属离子配位、细胞极性和蛋白质运输等领域具有互补的专业知识。这些高度合作的研究将解决对理解人类铜稳态至关重要的问题。具体地说，将结合蛋白质结晶、电子显微镜、X射线吸收光谱以及参与细胞内铜运输的分子(CTR1、CTR2、铜伴侣和铜运输ATPase)的功能分析来研究铜吸收和输出的分子机制。调控上皮细胞铜转运的因素将通过极化细胞系统、直接转运测量、共聚焦显微镜、定点突变和鉴定的蛋白质复合体的质谱学来表征。铜在正常组织和疾病组织中的生理调节方面将通过动物模型进行研究。该计划项目包括5个相互关联的研究计划以及金属离子核心。拟议的研究计划将产生新的信息，将数据整合到对人类铜稳态的多方面观点中，并将有助于更好地了解人类铜代谢障碍。将产生有用的共享试剂，并将提供给生物医学界。公共卫生相关性：PPG将解决与铜在人类生物学中的作用有关的基本问题，并将采用跨学科的方法来详细了解人类细胞中铜的动态平衡的分子机制。铜稳态失调是几种严重代谢紊乱的原因之一。拟议的研究将阐明人类细胞中铜调节的关键方面；这将有助于开发更好的铜相关疾病治疗方法。

DESCRIPTION (provided by applicant): The overall goal of this program is to generate a comprehensive mechanistic picture of human copper homeostasis in normal and diseased cells and tissues. Copper is an essential nutrient required for embryonic development, connective tissue formation, myelination of neurons, detoxification of radicals, and many other physiological processes. Copper deficiency is deleterious to human growth and development, yet excess copper is toxic. Dysregulation of copper homeostasis is associated with severe metabolic and neurological disorders, which may affect patients at any stage of life. In this Program Project, the mechanisms that control copper distribution in human cells and tissues will be characterized by a team of established investigators with complementary expertise in areas of structural biology, membrane transport, metal-ion coordination, cell polarity, and protein trafficking. The highly collaborative studies will address issues central to the understanding of human copper homeostasis. Specifically, the molecular mechanisms of copper uptake and export will be investigated using a combination of protein crystallization, electron microscopy, X-ray absorption spectroscopy as well as functional analysis of molecules involved in the intracellular copper trafficking (CTR1, CTR2, copper chaperones, and Cu-transporting ATPases). Factors that regulate copper transporters in epithelia will be characterized using polarized cell systems, direct transport measurements, confocal microscopy, site-directed mutagenesis, and mass-spectrometry of identified protein complexes. The physiological aspects of copper regulation in normal and diseased tissues will be investigated using animal models. The Program Project includes 5 interconnected research programs as well as the Metal Ion Core. The proposed research program will yield new information, integrate the data into a multifaceted view of human copper homeostasis, and will contribute to a better understanding of human disorders of copper metabolism. Useful shared reagents will be generated and will be made available to the biomedical community. Public Health Relevance: The PPG will address fundamental questions related to the role of copper in human biology and will employ an interdisciplinary approach to provide a detailed understanding of the molecular mechanisms of copper homeostasis in human cells. Dysregulation of copper homeostasis is a cause of several severe metabolic disorders. The proposed studies would elucidate the key aspects of copper regulation in human cells; that would facilitate the development of better therapies for copper-related disorders.