Biomedical Sciences, Defence Science and Technology Laboratory, Salisbury, Wiltshire SP4 OJQ, United Kingdom.
J Chromatogr B Analyt Technol Biomed Life Sci. 2010 May 15;878(17-18):1426-32. doi: 10.1016/j.jchromb.2009.11.030. Epub 2009 Nov 18.
Sulfur mustard (SM) is a potent vesicating agent that produces debilitating blisters and ulcerating lesions on the skin which are characteristically slow to heal. There are currently no specific medical countermeasures to prevent SM-induced vesication and therefore SM remains a major military threat. To investigate the mechanism by which SM causes these injuries we aimed to identify the cellular proteins that are important in the vesicant response and pathology of SM. Membrane and membrane-associated proteins that are targets for direct binding by SM were compared to targets directly bound by CEES (chloroethylethylsulphide). As CEES is a less potent blistering agent compared to SM, it was hypothesised that differences in the binding of these two mustards could reveal key proteins directly involved in the mustard vesicant response. Human cellular membranes fractionated from HaCaT cells were exposed to (14)C-SM or (14)C-CEES and the membrane proteins to which SM or CEES bound were separated by 2D gel electrophoresis, located by fluorography and subsequently identified using mass spectrometry. A number of proteins were identified that were differentially labelled by SM and CEES. Actin, annexin A2 and keratin 9 were labelled with SM at a higher intensity than was seen with the same concentration of CEES. Therefore results from these studies suggest that SM binding to these proteins could contribute to the complex pathology seen following SM exposure.
芥子气(SM)是一种有效的起疱剂,会在皮肤上产生使人衰弱的水疱和溃疡损伤,其愈合过程特别缓慢。目前尚无预防 SM 引起水疱的特殊医疗对策,因此 SM 仍然是一个主要的军事威胁。为了研究 SM 导致这些损伤的机制,我们旨在确定在 SM 致疱作用和病理学中起重要作用的细胞蛋白。将 SM 直接结合的膜和膜相关蛋白与 CEES(氯乙基乙基硫醚)直接结合的靶标进行了比较。由于 CEES 与 SM 相比是一种效力较弱的起疱剂,因此假设这两种芥子气结合的差异可以揭示直接参与芥子气致疱反应的关键蛋白。从 HaCaT 细胞中分离出的人细胞膜用 (14)C-SM 或 (14)C-CEES 暴露,用 2D 凝胶电泳分离与 SM 或 CEES 结合的膜蛋白,通过荧光显影定位,然后使用质谱法进行鉴定。鉴定出一些被 SM 和 CEES 差异标记的蛋白质。肌动蛋白、膜联蛋白 A2 和角蛋白 9 与 SM 的结合强度高于与相同浓度的 CEES 的结合强度。因此,这些研究的结果表明,SM 与这些蛋白的结合可能导致暴露于 SM 后出现的复杂病理。