Discovery of phosgene and chlorine gas modes of action and therapeutic targets using chemoproteomic profiling strategies

项目总结： 光气和氯是大规模工业生产中产生的两种有毒气体，会造成职业危害。 以及对公众和军方的化学威胁。这两种毒物都以不同的方式修饰蛋白质并诱导 肺部的氧化应激，它扰乱了关键的功能和途径。然而，准确的性质和 这些修饰在肺蛋白质组中的位置尚未确定，潜在的化学多样性 这些修饰对解开分子机制提出了分析挑战，通过这些机制 氯气和光气可致急性肺损伤。缺乏光气的生物分子特征--以及 因此，氯诱导的蛋白质靶标的修饰阻碍了我们开发有效的 对策：目前还没有获得批准的治疗光气或氯气的疗法 曝光。我们建议使用化学蛋白质组学--基于质谱学的蛋白质组学的子集 可以检测蛋白质的不同化学修饰的位置和类型-以全面识别 肺中被光气和氯化学改变的特定蛋白质和执行途径 分析以确定未来有针对性的对策开发的候选蛋白质。 这项建议的具体目标是： 目的1：动态氧化还原图谱以确定半胱氨酸氧化事件和全球蛋白质组 气态毒物暴露后的变化 目的2：用亲电化学方法定位光气和氯诱导的肺内蛋白质修饰 探头 目标3：数据集成和网络分析，以确定共享的和唯一的光气和 用于治疗开发的氯

Project Summary: Phosgene and chlorine are two toxic gases produced on large industrial scales that pose occupational hazards and chemical threats to the public and military. Both toxicants modify proteins in various ways and induce oxidative stress in the lung which perturb critical functions and pathways. However, the precise nature and sites of these modifications in the lung proteome have not been identified, and the potential chemical diversity of these modifications poses an analytical challenge to unraveling the molecular mechanisms by which chlorine and phosgene induce acute lung injury. This lack of biomolecular characterization of phosgene- and chlorine-induced modification of protein targets has consequently hindered our ability to develop effective countermeasures; there are currently no approved therapeutics for treating phosgene or chlorine gas exposures. We propose to use chemoproteomic profiling—a subset of mass spectrometry-based proteomics that can detect the sites and types of distinct chemical modifications to proteins—to comprehensively identify specific proteins in the lung that have been chemically altered by phosgene and chlorine and perform pathway analysis to identify protein candidates for future targeted countermeasure development. The specific aims of this proposal are: Aim 1: Dynamic redox profiling to identify the temporal protein cysteine oxidation events and global proteome changes following gaseous toxicant exposure Aim 2: Mapping phosgene- and chlorine-induced protein modifications in the lung using electrophilic chemical probes Aim 3: Data integration and network analysis to identify shared and unique molecular targets of phosgene and chlorine for therapeutic development