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Expression and characterization of a phospholipid hydroperoxide glutathione peroxidase gene in Schistosoma japonicum

机译：日本血吸虫中磷脂氢过氧化物谷胱甘肽过氧化物酶基因的表达与鉴定

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摘要

Phospholipid hydroperoxide glutathione peroxidase (PHGPx, GPx4) is a major antioxidant enzyme, which plays unique roles in the protection of cells against oxidative stress by catalysing reduction of lipid hydroperoxides. We isolated and characterized a full-length cDNA sequence encoding GPx gene from a blood fluke, Schistosoma japonicum (designated SjGPx), which contained an in-frame TGA codon for selenocysteine (Sec) and a concurrent Sec insertion sequence in its 3-untranslated region. Protein encoded by SjGPx demonstrated a primary structure characteristic to the PHGPx family, including preservation of catalytic domains and absence of the subunit interaction domains. Semi-quantitative reverse transcription PCR and Western blotting showed that the SjGPx was mainly expressed in the female adults and eggs. RNA interference approach was employed to investigate the effects of knockdown of SjGPx. SjGPx expression level was significantly reduced on the 5th day post-RNAi. Significantly reduction in GPx enzyme activities, as well as obvious changes in morphology of intrauterine eggs followed the reduction in SjGPx transcript level. We observed a 63.04% reduction in GPx activity and the eggs severely deformed. Our results revealed that SjGPx protein might be involved in the provision of enzyme activity during egg production.
机译：磷脂氢过氧化物谷胱甘肽过氧化物酶（PHGPx，GPx4）是一种主要的抗氧化酶，通过催化脂质氢过氧化物的还原，在保护细胞抵抗氧化应激中发挥独特作用。我们从血吸虫日本血吸虫（命名为SjGPx）中分离并鉴定了编码GPx基因的全长cDNA序列，其中包含硒代半胱氨酸（Sec）的读码框TGA密码子和在其3个非翻译区的并发Sec插入序列。由SjGPx编码的蛋白质显示了PHGPx家族的主要结构特征，包括催化结构域的保留和亚基相互作用域的缺失。半定量逆转录PCR和Western印迹表明SjGPx主要在成年雌性和卵中表达。 RNA干扰方法被用来研究SjGPx敲低的影响。 RNAi后第5天SjGPx表达水平显着降低。 SjGPx转录水平降低后，GPx酶活性显着降低，宫内卵的形态发生明显变化。我们观察到GPx活性降低了63.04％，鸡蛋严重变形。我们的结果表明，SjGPx蛋白可能参与产蛋过程中酶活性的提供。

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