METAL ION REGULATION CORE ADMINISTRATIVE PAGES

金属离子监管核心管理页面

• 批准号: 7690607
• 负责人: Martina Ralle
• 金额: $ 24.25万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-15 至 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7690607
• 关键词: AA Spectrophotometry; Antibody Formation; Antigens; Baculoviruses; Behavior; Businesses; Carrier Proteins; Categories; Cells; Charge; Communities; Confocal Microscopy; Copper; Core Facility; Data Analyses; Disease; Electrophoresis; Ensure; Equipment; Generations; Goals; Health; Homeostasis; Human; Human Resources; Imagery; Immunohistochemistry; Ion Transport; Ions; Mass Spectrum Analysis; Mediating; Metals; Microscope; Monoclonal Antibodies; Peptides; Protein Analysis; Proteins; Protocols documentation; Reagent; Recombinant Proteins; Regulation; Research; Research Personnel; Selenium; Selenocysteine; Services; Tissues; Training; Wilson disease protein; absorption; instrument; instrumentation; interest; mass spectrometer; mutant; programs; protein expression; research study; trafficking; two-dimensional

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)

金属离子核心的目的是提供仪器、技术支持和产生试剂 实现本应用程序的目标所必需的。核心将提供金属的原子吸收分析； 进行蛋白质双向电泳分析的设备和专业知识，一种质谱学 用于蛋白质和多肽分析的设备；以及用于免疫定位的共聚焦显微镜设备 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 研究铜的运输和转移所需的铜和硒离子(项目1、2和4)。弥撒 光谱仪将被用来识别与铜转运蛋白相互作用的蛋白质并调节其 细胞内定位和贩运行为(项目3、4和5)，以及确认 小角度x射线衍射实验制备的蛋白质中硒半胱氨酸的掺入。这个 共聚焦显微镜可以直接显示离子转运蛋白的亚细胞定位，从而 控制它们贩运正常和患病组织的机制是可以理解的(项目4)。这个 核心的重要任务将是表达和纯化用于产生抗体的重组蛋白 对抗参与铜稳态的蛋白质，并产生用于合作研究的蛋白质 在所有五个项目之间。这些后一类蛋白质包括ATP7B(项目5)、NBDDelta的各种突变体 TM 1，2(项目2和4)和Deltal-63蛋白质(项目4和5)