Yagi K, Komura S, Kojima H, Sun Q, Nagata N, Ohishi N, Nishikimi M
Institute of Applied Biochemistry, Yagi Memorial Parke, Mitake, Gifu, Japan.
Biochem Biophys Res Commun. 1996 Feb 15;219(2):486-91. doi: 10.1006/bbrc.1996.0260.
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.
通过聚合酶链反应（PCR）扩增从人睾丸互补脱氧核糖核酸（cDNA）中获得了编码人磷脂氢过氧化物谷胱甘肽过氧化物酶（PHGPx）的cDNA，并将其插入质粒pRc/CMV中，构建了人PHGPx的表达载体。用该表达载体转染豚鼠细胞系104C1细胞。其中一个转染子，命名为104Cl/O4C，对二亚油酰磷脂酰胆碱氢过氧化物和亚油酸氢过氧化物表现出高谷胱甘肽过氧化物酶活性。蛋白质印迹分析显示，转染子中有大量与抗PHGPx抗体发生免疫反应的蛋白质。当细胞与这些氢过氧化物一起孵育时，亲代细胞遭受严重的细胞损伤，而转染子对脂质氢过氧化物介导的损伤具有极强的抗性。
