METAL ION REGULATION CORE ADMINISTRATIVE PAGES

金属离子核心的目的是提供仪器，技术支持，并产生试剂 这是实现本申请目标所必需的。堆芯将提供金属原子吸收分析; 设备和专业知识，进行蛋白质的二维电泳分析，质谱 用于分析蛋白质和肽的设施;以及用于免疫定位的共聚焦显微镜设施， 细胞和组织中的蛋白质。原子吸收仪器将允许灵敏的定量分析 铜和硒离子研究铜的运输和转移（项目1，2和4）。质量 光谱仪将用于鉴定与铜转运蛋白相互作用的蛋白质，并调节它们的功能。 细胞内定位和运输行为（项目3，4和5），以及确认的效率 将硒代半胱氨酸掺入为小角X射线衍射实验制备的蛋白质中。的 共聚焦显微镜将允许离子转运蛋白的亚细胞定位的直接可视化， 可以理解控制它们在正常和病变组织中贩运的机制（项目4）。的 核心的重要任务是表达和纯化用于生产抗体的重组蛋白 针对参与铜稳态的蛋白质，并产生用于合作研究的蛋白质 五个项目之间。后一类蛋白质包括ATP 7 B的各种突变体（项目5）、NBD δ突变体（项目5）、NBD δ突变体（项目5）和NBD δ突变体（项目5）。 TMS 1、2（项目2和4）和deltal-63蛋白（项目4和5）

The purpose of the Metal Ion Core is to provide the instrumentation, technical support, and to generate reagents necessary to fulfill the goals of this application. The core will provide atomic absorption analysis of metals; equipment and expertise to perform two-dimensional electrophoresis analysis of proteins, a mass spectrometry facility for the analysis of proteins and peptides; and a confocal microscope facility for the immunolocalization of proteins in cells and tissues. The atomic absorption instrument will allow sensitive quantitative analysis of copper and selenium ions necessary to study copper transport and transfer (Projects 1,2, and 4). The mass spectrometer will be used to identify protein interacting with the copper transporters and regulating their intracellular localization and trafficking behavior (Projects 3,4, and 5), as well as to confirm the efficiency of incorporation of selenocysteine into proteins prepared for small angle x-ray diffraction experiments. The confocal microscope will allow direct visualization of the subcellular localization of ion transport proteins so that the mechanisms controlling their trafficking in normal and diseased tissue can be understood (Project 4). The important charge of the core will be to express and purify recombinant proteins for production of antibodies against proteins involved in copper homeostasis and well as to generate proteins for collaborative research between all five projects. These latter category of proteins includes various mutants of ATP7B (Project 5), NBDdelta TMS1,2 (Project 2 and 4), and deltal-63 protein (Projects 4 and 5)