METAL ION REGULATION CORE ADMINISTRATIVE PAGES

金属离子监管核心管理页面

• 批准号: 8137800
• 负责人: Martina Ralle
• 金额: $ 25.23万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8137800
• 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)

金属离子芯的目的是提供仪器，技术支持并生成试剂 实现此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； 对蛋白质进行二维电泳分析的设备和专业知识，质谱法 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对 研究铜运输和转移所需的铜和硒离子（项目1,2和4）。质量 光谱仪将用于鉴定蛋白质与铜转运蛋白相互作用并调节其 细胞内定位和贩运行为（项目3,4和5），并确认 将硒代半胱氨酸掺入为小角度X射线衍射实验制备的蛋白质。这 共聚焦显微镜将允许直接可视化离子转运蛋白的亚细胞定位，以便 可以理解控制其在正常组织和患病组织中贩运的机制（项目4）。这 核心的重要电荷将是表达和纯化重组蛋白以生产抗体 反对参与铜稳态的蛋白质，并生成蛋白质进行协作研究 在所有五个项目之间。这些后一种蛋白质包括ATP7B的各种突变体（项目5），NBDDELTA TMS1,2（项目2和4）和Deltal-63蛋白（项目4和5）