REACTIVE OXYGEN AND NITROGEN SPECIES IN RAW 2647 MACROPHAGE CELLS

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 氧化物种不仅介导了对各种病原体的静态和杀灭作用，而且在这种免疫反应中还参与了硝酸盐和氧化宿主蛋白的作用，并成为对处于慢性炎症状态的细胞内蛋白质进行翻译后修饰(PTM)的来源，这种慢性炎症状态称为衰老。我们用RPLC和FTICR MS研究了氧化应激后巨噬细胞蛋白质(特别是CaM)氧化和硝化修饰动力学的性质。我们发现，诱导巨噬细胞中自由基的产生刺激氧化和硝化CaM的清除，并选择性地从氧化修饰的CaM中切割C端赖氨酸。全面描述氧化应激(如炎症或衰老)对蛋白质修饰的变化，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体，将极大地加深我们对PTM是如何形成、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.