Glutathione S-transferase pi localizes in mitochondria and protects against oxidative stress.
Glutathione S-transferases (GSTs) are multifunctional enzymes involved in the protection of cellular components against anti-cancer drugs or peroxidative stress. Previously we found that GST pi, an isoform of the GSTs, is transported into the nucleus. In the present study, we found that GST pi is present in mitochondria as well as in the cytosol and nucleus in mammalian cell lines. A construct comprising the 84 amino acid residues in the amino-terminal region of GST pi and green fluorescent protein was detected in the mitochondria. The mutation of arginine to alanine at positions 12, 14, 19, 71, and 75 in full-length GST pi completely abrogated the ability to distribute in the mitochondria, suggesting that arginine, a positively charged residue, is required for the mitochondrial transport of GST pi. Chemicals generating reactive oxygen species, such as rotenone and antimycin A, decreased cell viability and reduced mitochondrial membrane potential. The overexpression of GST pi diminished these changes. GST pi-targeting siRNA abolished the protective effect of GST pi on the mitochondria under oxidative stress. The findings indicate that the peptide signal is conducive to the mitochondrial localization of GST pi under steady-state conditions without alternative splicing or posttranslational modifications such as proteolysis, suggesting that GST pi protects mitochondria against oxidative stress.
Goto
Shinji
S
Department of Biochemistry and Molecular Biology in Disease, Atomic Bomb Disease Institute, Nagasaki University Graduate School of Biomedical Sciences, Japan. sgoto@nagasaki-u.ac.jp
Kawakatsu
Miho
M
Izumi
Shin-Ichi
S
Urata
Yoshishige
Y
Kageyama
Kan
K
Ihara
Yoshito
Y
Koji
Takehiko
T
Kondo
Takahito
T
D000818:Animals; D000968:Antimycin A / Q000494:pharmacology; D001120:Arginine / Q000235:genetics; D019556:COS Cells; D002522:Chlorocebus aethiops; D019610:Cytoprotection / Q000187:drug effects / Q000235:genetics*; D004847:Epithelial Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D051549:Glutathione S-Transferase pi / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D008564:Membrane Potentials / Q000187:drug effects / Q000235:genetics; D008928:Mitochondria / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D016297:Mutagenesis, Site-Directed; D018384:Oxidative Stress / Q000187:drug effects; D021382:Protein Sorting Signals / Q000235:genetics; D021381:Protein Transport / Q000235:genetics*; D034741:RNA, Small Interfering / Q000235:genetics; D012402:Rotenone / Q000494:pharmacology; D014475:Uncoupling Agents / Q000494:pharmacology
D021382:Protein Sorting Signals; D034741:RNA, Small Interfering; D014475:Uncoupling Agents; D012402:Rotenone; D000968:Antimycin A; D001120:Arginine; D051549:Glutathione S-Transferase pi