Proteomic assessment of sulfur mustard-induced protein adducts and other protein modifications in human epidermal keratinocytes

作者：Mol Marijke A E^*; van den Berg Roland M; Benschop Henk P

来源：Toxicology and Applied Pharmacology, 2008, 230(1): 97-108.

DOI：10.1016/j.taap.2008.02.006

• sulfur mustard
• human epidermal keratinocytes
• cytokeratins
• protein alkylation
• caspase-6
• heat shock protein 27
• ribosomal protein P0
• stratifin
• galectin-7
• actin

Although some toxicological mechanisms of sulfur mustard (HD) have been uncovered, new knowledge will allow for advanced insight in the pathways that lead towards epidermal-dermal separation in skin. In the present investigation, we aimed to survey events that occur at the protein level in human epidermal keratinocytes (HEK) during 24 h after exposure to HID. By using radiolabeled 14 C-HID, it was found that proteins in cultured HEK are significant targets for alkylation by HID. HD-adducted proteins were visualized by two-dimensional gel electrophoresis and analyzed by mass spectrometry. Several type I and II cytokeratins, actin, stratifin (14-3-3 sigma) and galectin-7 were identified. These proteins are involved in the maintenance of the cellular cytoskeleton. Their alkylation may cause changes in the cellular architecture and, in direct line with that, be determinative for the onset of vesication. Furthermore, differential protcomic analysis was applied to search for novel features of the cellular response to HID. Partial breakdown of type I cytokeratins K14, K16 and K17 as well as the emergence of new charge variants of the proteins heat shock protein 27 and ribosomal protein PO were observed. Studies with caspase inhibitors showed that caspase-6 is probably responsible for the breakdown of type I cytokeratins in HEK. The significance of the results is discussed in terms of toxicological relevance and possible clues for therapeutic intervention.

• 出版日期2008-7-1