Chlorinated lipid modification of proteins: Biomarkers of chlorine gas exposure

暴露于氯气（Cl 2）对公众健康构成重大威胁。氯是一种主要的化学物质 在美国大量生产。暴露于Cl 2是由于火车出轨、意外事故 游泳池维修工人的滥用、水处理设施的事故和化学战。 故意暴露是一个主要问题，因为Cl 2既是化学战剂，也是潜在的化学品 恐怖分子需要诊断生物标志物来确定人类暴露于Cl 2的程度， 预测结果，这最终可能导致改善治疗支持和对策。福特 实验室发现，Cl 2和HOCl都靶向宿主血浆脂质，导致2-氯脂肪醛（2-CLFAD）和2-氯脂肪酸（2-CLFA）的产生。这些氯化脂质积累到强大的水平， 肺和血浆的小鼠暴露于亚致死量的氯气。在最近未发表的研究中， 显示在暴露于氯的5个受试者队列中，血浆2-CLFA水平升高40倍， 亚拉巴马伯明翰附近的水处理设施。总之，我们已经证明氯化脂质来源于 是目前人类暴露于Cl 2的最佳生物标志物， 有可能用于预测未来的结果，从而改善暴露后的护理。血浆 使用LC/MS检测，采用三重四极杆仪器对氯化脂质进行定量。的 对LC/MS检测的依赖是使用这些方法进行Cl 2暴露现场分析的局限性。 生物标志物。拟议的研究将通过鉴定氯脂质修饰的蛋白质来解决这个问题 产生于Cl 2暴露之后。未来的研究生产抗体氯脂质修饰的蛋白质， 然后用于开发诊断试剂盒。有人提出，氯化脂质共价修饰蛋白质。 氯化脂质修饰的特定蛋白质和氨基酸残基的鉴定将用于 未来的研究将开发可用于快速检测分析的抗体。此外，据设想， 识别氯脂质修饰的蛋白质作为Cl 2暴露的结果可能为未来的研究提供新的目标。 调查研究，以制定应对Cl 2暴露的对策。有三个具体目标。 Specific Aim 1将鉴定由2-CLINID修饰的氨基酸基序和蛋白质。 Specific Aim 2将鉴定由2-CLFA修饰的氨基酸基序和蛋白质。 具体目标3将识别暴露于Cl 2的小鼠肺和血浆中的氯化脂质修饰蛋白。 这些研究将鉴定氯化脂质修饰的蛋白质，并表明它们存在于Cl 2的血浆中。 暴露的老鼠此外，这些拟议的R21研究有可能导致在 未来将开发改进的对策，并改善结果，Cl 2暴露。

Exposure to chlorine gas (Cl2) presents a significant threat to public health. Chlorine is a leading chemical produced by volume in the US. Exposure to Cl2 has occurred as a result of train derailments, accidental misuse by swimming pool maintenance workers, accidents at water treatment facilities, and chemical warfare. Intentional exposure is a major concern since Cl2 is both a chemical warfare agent and potential chemical terror agent. Diagnostic biomarkers are needed to determine the extent of Cl2 exposure to humans and to predict outcomes, which ultimately could lead to improved therapeutic support and countermeasures. The Ford lab discovered that both Cl2 and HOCl target host plasmalogen lipids, resulting in 2-chlorofatty aldehyde (2-CLFALD) and 2-chlorofatty acid (2-CLFA) production. These chlorinated lipids accumulate to robust levels in lung and plasma of mice exposed to sub-lethal amounts of chlorine gas. In recent unpublished studies we have shown plasma 2-CLFA levels are elevated 40-fold in a five-subject cohort of humans exposed to chlorine at a water treatment facility near Birmingham, Alabama. Taken together, we have shown chlorinated lipids derived from plasmalogen oxidation are currently the best biomarkers of Cl2 exposure to humans and have the potential to be used to predict future outcomes that could lead to improved care following exposure.!Plasma chlorinated lipids are quantified using LC/MS detection employing a triple quadrupole instrument. The dependence on LC/MS detection is a limitation in implementing field analyses of Cl2 exposure using these biomarkers. The proposed studies will address this problem by identifying chlorolipid-modified proteins produced following Cl2 exposure. Future studies producing antibodies to chlorolipid-modified proteins could then be used to develop diagnostic kits. It is proposed that chlorinated lipids covalently modify proteins. The identification of specific proteins and amino acid residues modified by chlorinated lipids will be used in future studies to develop antibodies that can be used for rapid detection assays. Additionally, it is envisioned that identifying chlorinated lipid-modified proteins as a result of Cl2 exposure may provide new targets for future investigations to develop countermeasures to Cl2 exposure. There are three specific aims. Specific Aim 1 will identify amino acid motifs and proteins modified by 2-CLFALD. Specific Aim 2 will identify amino acid motifs and proteins modified by 2-CLFA. Specific Aim 3 will identify chlorinated lipid-modified proteins in lung and plasma from mice exposed to Cl2. These studies will identify chlorinated lipid-modified proteins, and show that they are present in plasma of Cl2 exposed mice. Furthermore, these proposed R21 studies have the potential to lead to new investigations in the future to develop improved countermeasures for, and improve outcomes of, Cl2 exposure.