Expression of human phospholipid hydroperoxide glutathione peroxidase gene for protection of host cells from lipid hydroperoxide-mediated injury.
A cDNA encoding human phospholipid hydroperoxide glutathione peroxidase (PHGPx) was obtained by PCR amplification from human testis cDNA and was inserted into the plasmid pRc/CMV to construct an expression vector for human PHGPx. Guinea pig cell line 104C1 cells were transfected with the expression vector. One of the transfectants, designated 104Cl/O4C, expressed high glutathione peroxidase activity toward dilinoleoyl phosphatidylcholine hydroperoxide and linoleic acid hydroperoxide. Western blot analysis revealed a large amount of protein immunoreactive against anti-PHGPx antibody in the transfectant. When the cells were incubated with these hydroperoxides, the parental cells suffered from serious cell injury, whereas the transfectant was extremely resistant against lipid hydroperoxide-mediated injury.
Yagi
K
K
Institute of Applied Biochemistry, Yagi Memorial Parke, Mitake, Gifu, Japan.
Komura
S
S
Kojima
H
H
Sun
Q
Q
Nagata
N
N
Ohishi
N
N
Nishikimi
M
M
D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects*; D018076:DNA, Complementary; D015870:Gene Expression / Q000187:drug effects*; D005979:Glutathione Peroxidase / Q000096:biosynthesis* / Q000235:genetics; D006168:Guinea Pigs; D006801:Humans; D008041:Linoleic Acids / Q000633:toxicity*; D008054:Lipid Peroxides / Q000633:toxicity*; D008297:Male; D008969:Molecular Sequence Data; D010713:Phosphatidylcholines / Q000633:toxicity*; D000080662:Phospholipid Hydroperoxide Glutathione Peroxidase; D010957:Plasmids; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000096:biosynthesis; D013737:Testis / Q000201:enzymology; D014162:Transfection
D018076:DNA, Complementary; D008041:Linoleic Acids; D008054:Lipid Peroxides; D010713:Phosphatidylcholines; D011994:Recombinant Proteins; C057590:phosphatidylcholine hydroperoxide; C008285:linoleic acid hydroperoxide; D000080662:Phospholipid Hydroperoxide Glutathione Peroxidase; D005979:Glutathione Peroxidase