REACTIVE OXYGEN AND NITROGEN SPECIES IN RAW 2647 MACROPHAGE CELLS

原始 2647 巨噬细胞中的活性氧和氮物种

• 批准号: 7602874
• 负责人: THOMAS Comey SQUIER
• 金额: $ 4.2万
• 依托单位: BATTELLE PACIFIC NORTHWEST LABORATORIES
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-07-01 至 2008-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7602874
• 关键词: Affect; Aging; C-terminal; Calmodulin; Cell physiology; Cells; Chronic; Cleaved cell; Complex; Computer Retrieval of Information on Scientific Projects Database; Coupled; Funding; Generations; Grant; Immune response; Inflammation; Institution; Lysine; Mediating; Modification; Nature; Nitrates; Nitrogen; Oxidative Stress; Oxygen; Post-Translational Protein Processing; Proteins; Research; Research Personnel; Resources; Source; United States National Institutes of Health; macrophage; nitrate; pathogen; repaired

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Oxidative species not only mediate static and cidal effects on a variety of pathogens, but also nitrate and oxidize host proteins during this immune response, and become a source of posttranslational modifications (PTMs) to intracellular proteins in the chronic inflammation state called aging. We have investigated the nature of oxidative and nitrative modification dynamics on macrophage proteins (specifically calmodulin, CaM) after oxidative stress using RPLC coupled with FTICR MS. We showed that induced radical generation in macrophages stimulates a clearance of oxidized and nitrated CaM and selectively cleaves the C-terminal lysine from oxidatively modified CaM. Comprehensive characterization of the changes in modifications to proteins with oxidative stress (e.g. inflammation or aging) and the discovery of how the modifications affect the protein-protein interface, and by extension alter protein complexes, will greatly further our understanding of how PTMs are formed, distributed, repaired and otherwise interfere with normal cellular processes.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目及 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 氧化物质不仅介导对多种病原体的静态和杀伤作用，而且在免疫反应过程中硝酸盐和氧化宿主蛋白，并成为称为衰老的慢性炎症状态下细胞内蛋白翻译后修饰（PTM）的来源。我们使用 RPLC 与 FTICR MS 联用研究了氧化应激后巨噬细胞蛋白（特别是钙调蛋白，CaM）氧化和硝基修饰动力学的性质。我们发现巨噬细胞中诱导的自由基生成会刺激氧化和硝化 CaM 的清除，并选择性地从氧化修饰的 CaM 中裂解 C 末端赖氨酸。全面表征氧化应激（例如炎症或衰老）引起的蛋白质修饰变化，以及发现这些修饰如何影响蛋白质-蛋白质界面，进而改变蛋白质复合物，将极大地加深我们对 PTM 如何形成的理解，分布、修复或以其他方式干扰正常细胞过程。