Mammalian small stress proteins protect against oxidative stress through their ability to increase glucose-6-phosphate dehydrogenase activity and by maintaining optimal cellular detoxifying machinery.
The protective activity of small stress proteins (sHsp) against H2O2-mediated cell death in the highly sensitive murine L929 fibroblast has been analyzed. We report here that the human Hsp27- and murine Hsp25-mediated rise in glutathione (GSH) levels as well as the maintenance of this redox modulator in its reduced form was directly responsible for the protection observed at the level of cell morphology and mitochondrial membrane potential. sHsp expression also buffered the increase in protein oxidation following H2O2 treatment and protected several key enzymes against inactivation. In this case, however, the protection necessitated both an increase in GSH and the presence of sHsp per se since the pattern of protection against protein oxidation mediated by a simple GSH increase was different from that induced by sHsp expression. Among the enzymes analyzed, we noticed that sHsp significantly increased glucose-6-phosphate dehydrogenase (G6PD) activity and to a lesser extent glutathione reductase and glutathione transferase activities. Moreover, an increased GSH level was observed in G6PD-overexpressing L929 cell clones. Taken together our results suggest that sHsp protect against oxidative stress through a G6PD-dependent ability to increase and uphold GSH in its reduced form and by using this redox modulator as an essential parameter of their in vivo chaperone activity against oxidized proteins.
Préville
X
X
Centre de Génétique Moléculaire et Cellulaire, CNRS-UMR-5534, Villeurbanne, F-69622, France.
Salvemini
F
F
Giraud
S
S
Chaufour
S
S
Paul
C
C
Stepien
G
G
Ursini
M V
MV
Arrigo
A P
AP
D000818:Animals; D000975:Antioxidants / Q000378:metabolism* / Q000494:pharmacology*; D002460:Cell Line; D004789:Enzyme Activation / Q000187:drug effects; D005434:Flow Cytometry; D005954:Glucosephosphate Dehydrogenase / Q000378:metabolism*; D006360:Heat-Shock Proteins / Q000096:biosynthesis / Q000502:physiology*; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D007425:Intracellular Membranes / Q000502:physiology; D008564:Membrane Potentials / Q000502:physiology; D051379:Mice; D008928:Mitochondria / Q000502:physiology; D010084:Oxidation-Reduction; D018384:Oxidative Stress; D014407:Tumor Cells, Cultured
D000975:Antioxidants; D006360:Heat-Shock Proteins; D006861:Hydrogen Peroxide; D005954:Glucosephosphate Dehydrogenase