REACTIVE OXYGEN AND NITROGEN SPECIES IN RAW 2647 MACROPHAGE CELLS

该子项目是利用 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH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 中心，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灭作用，而且在这种免疫应答期间硝酸化和氧化宿主蛋白，并成为称为老化的慢性炎症状态中细胞内蛋白的翻译后修饰（PTM）的来源。我们已经调查了氧化和硝化修饰动力学的性质，对巨噬细胞蛋白（特别是钙调素，钙调素）氧化应激后，使用RPLC加上FTICR MS。我们表明，诱导的自由基产生在巨噬细胞刺激清除氧化和硝化钙调素，并选择性地切割C-末端赖氨酸从氧化修饰钙调素。对具有氧化应激（例如炎症或老化）的蛋白质的修饰的变化的全面表征以及对修饰如何影响蛋白质-蛋白质界面以及通过延伸改变蛋白质复合物的发现，将大大促进我们对PTM如何形成、分布、修复以及以其他方式干扰正常细胞过程的理解。

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.