Redox Molecular Signaling Core

项目摘要 COBRE氧化还原生物学和心血管疾病中心的目标是建立一个全国性的 公认的先进的研究和培训计划，在氧化还原生物调节心血管疾病， 促进初级教师科研项目的发展，实现重大科研项目的自主资助。 氧化还原分子信号核心（核心C）为COBRE研究人员和学员提供了访问 用于检测和定量活性氧物质，活性氮物质， 和反应性硫物质。此外，该设备还可以分析蛋白质的氧化还原依赖性控制 功能通过氧化还原蛋白质组学和代谢组学，提供了一个集中的设施，细胞培养缺氧 研究，并为建立氧化还原信号的细胞培养模型提供专业知识。这一系列分析 技术来评估氧化还原信号将是无与伦比的一个单一的设施在路易斯安那州。的职能 氧化还原分子信号核心是通过两个不同的子核心，分析氧化还原生物学 子核心和分子信号子核心。分析氧化还原生物学子核心利用设备， 高效液相色谱法（HPLC）、专用荧光检测器和质谱仪 质谱仪（MS），以提供高品质，定量氧化还原测量在一个集中的设施， 敬业的技术人员。在第一阶段，该机构购买了一台Orbitrap Exploris 480质谱仪， 这显著地扩展了我们在模型中对细胞和组织功能进行蛋白质组学分析的能力， 氧化还原信号在第二阶段，我们将通过购买新的 用于活性硫物质定量的设备和额外的质谱设备， 灵敏度分子信号传递子核心提供了一个集中的设施，用于提供分子信号传递的帮助。 生物学，用于建立氧化还原信号传导的细胞培养模型，并为获得重要的设备， 使用高通量自动化毛细管模拟培养中的缺氧细胞损伤并用于蛋白质分析 电泳在第二阶段，我们将通过增加额外的分子信号传导子核心， 基于当前COBRE教师需求的核心服务，例如改进的瞬态方法 原代细胞的转染并增强对下一代测序技术的利用。教职 氧化还原分子信号核心的工作人员与COBRE研究人员密切合作，以优化实验 设计、执行分析测量、提供核心设备使用方面的培训，并协助处理数据 收集和解释。这些服务为COBRE调查人员提供了宝贵的好处， 在加强当地氧化还原生物学研究基础设施的同时，推进他们的个别项目。

PROJECT ABSTRACT The goals of the COBRE Center for Redox Biology and Cardiovascular Disease are to establish a nationally recognized advanced research and training program in redox biology regulation of cardiovascular disease and to facilitate the development of junior faculty research programs to achieve major independent research funding. The Redox Molecular Signaling Core (Core C) provides COBRE investigators and trainees with access to modern equipment for the detection and quantification of reactive oxygen species, reactive nitrogen species, and reactive sulfur species. Additionally, this facility enables the analysis of redox-dependent control of protein function through redox proteomics and metabolomics, provides a centralized facility for cell culture hypoxia studies, and provides expertise for establishing cell culture models of redox signaling. This range of analytical techniques to assess redox signaling will be unmatched for a single facility in Louisiana. The functions of the Redox Molecular Signaling Core are accomplished through two distinct sub-cores, the Analytical Redox Biology Sub-Core and the Molecular Signaling Sub-Core. The Analytical Redox Biology Sub-Core utilizes equipment for high performance liquid chromatography (HPLC), specialized chemiluminescent detectors, and mass spectrometry (MS) to provide high quality, quantitative redox measurements at a centralized facility with dedicated technical staff. During Phase I, the institution purchased an Orbitrap Exploris 480 mass spectrometer that significantly expanded our capability to perform proteomic analysis of cell and tissue function in models of redox signaling. During Phase II, we will expand the functionality of this core through the purchase of new equipment for reactive sulfur species quantification and additional mass spectrometry equipment to enhance sensitivity. The Molecular Signaling Sub-Core offers a centralized facility for providing assistance with molecular biology, for establishing cell culture models of redox signaling, and for affording access to vital equipment for modeling hypoxic cell injury in culture and for protein analysis using high throughput, automated capillary electrophoresis. In Phase II, we will mature and expand the Molecular Signaling Sub-Core by adding additional core services based on the needs of the current COBRE faculty, such as improved methods for transient transfection of primary cells and enhanced access to next generation sequencing technologies. The faculty and staff of the Redox Molecular Signaling Core work closely with the COBRE investigators to optimize experimental design, perform analytical measurements, provide training on the use of core equipment, and assist with data collection and interpretation. These services provide an invaluable benefit to the COBRE investigators to advance their individual projects while enhancing the infrastructure of local redox-biology research.