Journal
FREE RADICAL BIOLOGY AND MEDICINE
Volume 84, Issue -, Pages 215-226Publisher
ELSEVIER SCIENCE INC
DOI: 10.1016/j.freeradbiomed.2015.02.032
Keywords
Peroxiredoxin-5; Peroxisomes; Mitochondria; Oxidative stress; Killer Red; Lipid peroxidation; roGFP2; Free radicals
Funding
- Fonds pour la formation A la Recherche dans l'Industrie et dans l'Agriculture
- DIANE research program of the Walloon region [10/15-026]
- FWO [G.0754.09]
- Bijzonder Onderzoeksfonds van de K.U. Leuven [DBOF/10/059, OT/14/100]
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Peroxiredoxin-5 (PRDX5) is a thioredoxin peroxidase that reduces hydrogen peroxide, alkyl hydroperoxides, and peroxynitrite. This enzyme is present in the cytosol, mitochondria, peroxisomes, and nucleus in human cells. Antioxidant cytoprotective functions have been previously documented for cytosolic, mitochondrial, and nuclear mammalian PRDX5. However, the exact function of PRDX5 in peroxisoines is still not clear. The aim of this work was to determine the function of peroxisomal PRDX5 in mammalian cells and, more specifically, in glial cells. To study the role of PRDX5 in peroxisomes, the endogenous expression of PRDX5 in murine oligodendrocyte 158 N cells was silenced by RNA interference. In addition, human PRDX5 was also overexpressed in peroxisomes using a vector coding for human PRDX5, whose unconventional peroxisomal targeting sequence 1 (PTS1; SQL) was replaced by the prototypical PTS1 SKL. Stable 158 N clones were obtained. The antioxidant cytoprotective function of peroxisomal PRDX5 against peroxisomal and mitochondrial KillerRed-mediated reactive oxygen species production as well as H2O2 was examined using MTT viability assays, roGFP2, and C11-BOBIPY probes. Altogether our results show that peroxisomal PRDX5 protects 158 N oligodendrocytes against peroxisomal and mitochondrial KillerRed- and H2O2-induced oxidative stress. (C) 2015 Elsevier Inc. All rights reserved.
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