Differential correlations between changes to glutathione redox state, protein ubiquitination, and stress-inducible HSPA chaperone expression after different types of oxidative stress

摘要: In primary bovine fibroblasts with an hspa1b/luciferase transgene, we examined the intensity of heat-shock response (HSR) following four types oxidative stress or heat (HS), and its putative relationship changes to different cell parameters, including reactive oxygen species (ROS), redox status key molecules glutathione (GSH), NADP(H) NAD(H), post-translational protein modifications carbonylation, S-glutathionylation, ubiquitination. We determined sub-lethal condition generating maximal luciferase activity inducible HSPA level for treatments hydrogen peroxide (H2O2), UVA-induced photo-activation, superoxide-generating agent menadione (MN), diamide (DA), electrophilic sulfhydryl reagent. The HSR induced by was highest after DA MN, followed UVA H2O2 treatments, not correlated ROS production nor extent S-glutathionylation carbonylation observed immediately stress. found a correlation between GSH/GSSG stress, increase in ubiquitination during recovery period. Conversely, HS treatment, which led level, did generate modified depended on GSH state. Furthermore, maximum lower than MN thereafter. these cells, heat-induced therefore clearly from stress-induced HSR, conversely early major cellular thiol predicted polyubiquinated proteins.