Mechanisms of Metal Ion Regulation in Human Cells

修订摘要：该项目资助的总体目标是阐明人类细胞中金属稳态的机制。铜和铁是各种生物过程中必不可少的辅助因子，虽然这些金属的低水平是必不可少的，但高水平是有毒的。铜的生理处理可以分为几个步骤。这些是，摄取到细胞中，传递到特定的铜依赖性蛋白质和从细胞中去除。铜分布的分子机制将在分子、细胞和组织水平上以跨学科的方法进行研究。 在人类中，摄取是由hCtrl介导的，PPG的一个项目将对这种最近发现的蛋白质进行详细的表征。第二个PPG项目将通过铜分子伴侣对铜依赖蛋白进行特异性递送的结构和机制分析。 铜的去除是通过铜激活的ATP酶完成的，这些ATP酶在几个水平上受到铜的调节。在项目3中，最近发现的这些ATP酶之一的铜依赖性磷酸化将被用作理解铜信号传导和调节的基础的模型。PPG核心包括共聚焦显微镜、质谱仪和其他仪器，促进了这种多级策略。 金属稳态的破坏越来越多地被援引为许多神经退行性疾病和其他危及生命的疾病的基础。为了发现新的和更有效的治疗方法，有必要更清楚地了解正常状态下金属离子的调节。该计划的目标是启动对这些过程的系统调查，并向生物医学界提供这些信息。

REVISED ABSTRACT: The overall goal of this Program Project Grant is to elucidate the mechanisms involved in the metal homeostasis by human cells. Copper and iron are essential co-factors in a wide variety of biological processes and while low regulated levels of these metals are essential, elevated levels are toxic. The physiological handling of copper can be viewed in several steps. These are, uptake into the cell, delivery to specific copper-dependent proteins and removal from the cell. The molecular mechanisms of copper distribution will be studied at the molecular, cellular and tissue level in an interdisciplinary approach. Uptake, in humans, is mediated by hCtrl, and one of the PPG projects will carry out a detailed characterization of this recently discovered protein. The second PPG project will perform a structural and mechanistic analysis of specific delivery to copper dependent proteins by copper chaperones. The removal of copper is accomplished by copper-activated ATPases that are regulated by copper at several levels. In Project 3, a recently discovered copper-dependent phosphorylation of one of these ATPases will be used as a model to understand the basis of copper signaling and regulation. The PPG core including confocal microscopy, mass spectrometer, and other instrumentation, facilitates this multi-level strategy. Disruptions in metal homeostasis are being increasingly invoked as the basis for many neurodegenerative and other life-threatening diseases. In order that new and more effective therapies can be discovered it is necessary to have a clearer understanding of the regulation of metal ions in normal states. The goal of this program is to initiate a systematic investigation of these processes and provide such information to the biomedical community.