Proteome analysis of glial cells treated by radiation or hydroperoxide

Reactive oxygen species (ROS) give rise to various types of oxidative damage and some responses such as activation of transcription factors and/or repair factors in cells. Proteome analysis using 2-D PAGE and peptide mass fingerprinting is suitable for the study on the post-translational modifications of proteins. When glial cells were exposed to hydroperoxide, the relative abundance of 9 spots changed on 2-D gels, as compared with control gels. MALDI-TOF MS analysis after in-gel digestion revealed that these spots corresponded to at least 3 pairs of proteins. These pairs of protein spots had different isoelectric points each other and were identified as peroxiredoxin II, peroxiredoxin III and calpactin I light chain. It was demonstrated that peroxiredoxins II and III were oxidized to their acidic forms (acidic spots) by hydroperoxide, and thereafter the native forms (basic spots) were recovered as the acidic forms disappeared.