Copper entry into human cells

铜是一种必需的营养素，是许多不同的细胞过程所必需的。它作为抗氧化防御、神经递质生物合成和许多其他重要生化转化中的辅助因子是重要的。虽然低细胞水平的存在对人类健康至关重要，但水平升高是有毒的。因此，铜从细胞和组织进入和消除是非常重要的途径。最近，负责介导铜进入人体细胞的蛋白质已被鉴定，并被称为hCtr 1。这种完整的膜蛋白由190个氨基酸残基组成，并出现（从亲水性分析），像它的酵母同系物跨越膜三次。它已被证明是正常胚胎发育所必需的。本申请的PI最近 在用含有hCtr 1的eDNA的杆状病毒颗粒感染的昆虫细胞中成功地实现了hCtr 1的异源表达。本申请旨在提供hCtr 1的结构-功能关系的第一个详细表征，并将研究由hCtr 1介导的铜转运机制。这些研究将把同位素铜摄取的联合收割机功能测定与新型hCtr 1分子的诱变分析结合起来。这些研究将通过使用共聚焦显微镜和PI实验室生产的特异性Ab检查哺乳动物细胞中内源性hCtr 1水平的运输来增强。这些研究的结果将为理解人类铜调节的一个重要方面提供一个重要的起点。铜代谢和体内平衡的破坏与几种神经退行性疾病有关，新的治疗策略的开发需要更多地了解正常和疾病状态下的调节途径。

Copper is an essential nutrient and is required by a number of distinct cellular processes. It is important as a co-factor in anti-oxidant defense, in neurotransmitter biosynthesis and a number of other important biochemical transformations. Although the presence of low cellular levels is essential for human health, elevated levels are toxic. Thus, copper entry and elimination from cells and tissues are highly important pathways. Recently the protein responsible for mediating the entry of copper into human cells has been identified and is called hCtr1. This integral membrane protein consists of 190 arninoacid residues and appears (from hydropathy analysis), like its yeast homologues to span the membrane three times. It has been demonstrated to be essential for normal embryonic develpment. The PI of this application has recently achieved the successful heterologous expression of hCtr1 in insect cells infected with baculovirus particles containing the eDNA for hCtr1. The present application aims to provide the first detailed characterization of structure-function relations in hCtr1 and will investigate the mechanism of copper transport mediated by hCtr1. The studies will combine functional assays of isotopic copper uptake with mutagenic analysis of novel hCtr1 molecules. These studies will be augmented by an examination of trafficking of endogenous levels of hCtr1 in mammalian cells using confocal microscopy and specific Ab's produced by the PI's laboratory. The results of these studies will provide an essential starting point for an understanding of an important aspect of copper regulation in humans. Disruptions in copper metabolism and homeostasis have been associated with several neurodegenerative conditions and the development of new therapeutic strategies requires greater knowledge of the regulatory pathways in normal and diseased states.