Molecular expression and enzymatic characterization of thioredoxin from the carcinogenic human liver fluke Opisthorchis viverrini.
Molecular expression and enzymatic characterization of thioredoxin from the carcinogenic human liver fluke Opisthorchis viverrini.
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致癌人肝吸虫 Opisthorchis viverrini 硫氧还蛋白的分子表达和酶学特征。
DOI：
10.1016/j.parint.2011.06.018
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发表时间：
2012
影响因子：
1.9
通讯作者：
Sripa,Banchob
中科院分区：
文献类型：
--
作者：
Suttiprapa,Sutas;Matchimakul,Pitchaya;Loukas,Alex;Laha,Thewarach;Wongkham,Sopit;Kaewkes,Sasithorn;Brindley,PaulJ;Sripa,Banchob
人类肝吸虫，猫尾后睾吸虫，诱导肝胆系统的炎症。尽管经常暴露在炎症细胞释放的有害氧自由基中，寄生虫仍能存活多年。对氧化损伤的防御可以通过谷胱甘肽和/或硫氧还蛋白利用系统介导。在这里，我们报告的分子表达和生化特性的硫氧还蛋白（Trx）从O。维韦里尼。O. ViverriniTrx cDNA编码105个氨基酸残基的多肽，分子量为11.63kDa。预测的蛋白质与先前表征的硫氧还蛋白具有26-51%的相似性。重组O. Viverrini Trx（Ov-Trx-1）在E.杆菌重组蛋白具有胰岛素还原活性，并支持O.灵猫硫氧还蛋白过氧化物酶。Ov-Trx-1在mRNA和蛋白质水平的表达，观察在所有可获得的发育阶段的肝吸虫。Ov-Trx-1也在O.维韦里尼。免疫组化结果显示，Ov-Trx-1在除卵巢和成熟精子外的所有寄生虫组织中均有表达。有趣的是，Ov-Trx-1在感染的胆管上皮中观察到，但在正常胆管中没有。这些结果表明Ov-Trx-1在寄生虫的整个生命周期中是必不可少的。在宿主-寄生虫相互作用方面，Ov-Trx-1可能支持硫氧还蛋白过氧化物酶保护寄生虫免受由活性氧引起的炎症损伤。
The human liver fluke, Opisthorchis viverrini, induces inflammation of the hepatobiliary system. Despite being constantly exposed to inimical oxygen radicals released from inflammatory cells, the parasite survives for years. Defense against oxidative damage can be mediated through glutathione and/or thioredoxin utilizing systems. Here, we report the molecular expression and biochemical characterization of a thioredoxin (Trx) from O. viverrini. O. viverrini Trx cDNA encoded a polypeptide of 105 amino acid residues, of molecular mass 11.63kDa. The predicted protein has similarity to previously characterized thioredoxins with 26–51% identity. Recombinant O. viverrini Trx (Ov-Trx-1) was expressed as soluble protein in E. coli. The recombinant protein showed insulin reduction activity and supported the enzymatic function of O. viverrini thioredoxin peroxidase. Expression of Ov-Trx-1 at mRNA and protein levels was observed in all obtainable developmental stages of the liver fluke. Ov-Trx-1 was also detected in excretory–secretory products released by adult O. viverrini. Immunohistochemistry, Ov-Trx-1 was expressed in nearly all parasite tissue excepted ovary and mature sperms. Interestingly, Ov-Trx-1 was observed in the infected biliary epithelium but not in normal bile ducts. These results suggest that Ov-Trx-1 is essential for the parasite throughout the life cycle. In the host–parasite interaction aspect, Ov-Trx-1 may support thioredoxin peroxidase in protecting the parasite against damage induced by reactive oxygen species from inflammation.