Molecular cloning and characterization of peroxiredoxin 4 involved in protection against oxidative stress in the silkworm Bombyx mori

Molecular cloning and characterization of peroxiredoxin 4 involved in protection against oxidative stress in the silkworm Bombyx mori
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参与家蚕抗氧化应激的过氧化还原蛋白 4 的分子克隆和表征

DOI:
10.1111/j.1365-2583.2012.01161.x
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发表时间:
2012-12
期刊:
Insect Mol Biol.
影响因子:
--
通讯作者:
Z Zhang
Z Zhang
中科院分区:
其他
文献类型:
--
作者:
Shi GQ;QY Yu;L Shi;Z Zhang

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过氧化还原蛋白 (Prxs) 是一个普遍存在的蛋白质家族,通过细胞过氧化物的解毒,在抵御氧化应激方面发挥着重要作用。在这里,我们描述了家蚕 Bombyx mori (BmPrx4) 的 Prx4 cDNA 的克隆和表征。 BmPrx4 基因有一个 744bp 的开放阅读框,编码 248 个氨基酸和一个保守基序 VCP,参与其假定的氧化还原功能。该基因在大肠杆菌中的异源表达蛋白表现出抗氧化活性,去除过氧化氢并保护 DnA。蛋白质印迹分析显示血淋巴中存在 BmPrx4,表明该蛋白质是可分泌的。此外,BmPrx4 在所有发育阶段都有表达。 BmPrx4在摄食阶段表达量较低,但在游走阶段表达量较高。 BmPrx4 是由槲皮素或温度应激诱导的。免疫组织化学分析表明,BmPrx4 存在于蚕的大脑、神经元和头部嗅觉器官中。总的来说,我们的结果表明 BmPrx4 的表达谱与氧化损伤的保护密切相关。蚕是鳞翅目害虫的代表,我们的数据为农林害虫防治技术的发展提供了线索。
Peroxiredoxins (Prxs) are a ubiquitous family of proteins that play important roles in insects in protection against oxidative stress through the detoxification of cellular peroxides. Here, we describe the cloning and characterization of a Prx4 cDNA of the silkworm Bombyx mori (BmPrx4). The BmPrx4 gene has an open reading frame of 744 bp encoding 248 amino acids and a conserved motif, VCP, involved in its presumed redox functions. The heterologously expressed proteins of the gene in Escherichia coli showed antioxidant activity, removed hydrogen peroxide and protected DnA. Western blotting analysis showed the presence of BmPrx4 in the haemolymph, suggesting that the protein is secretable. Moreover, BmPrx4 was expressed at all developmental stages. The expression level of BmPrx4 was relatively low during the feeding stage but high at the wandering stage. BmPrx4 was induced by quercetin or temperature stress. Immunohistochemical analysis revealed that BmPrx4 is present in the brain, neurones and olfactory organ of the head in silkworms. Overall, our results indicate that the expression profile of BmPrx4 correlates well with protection from oxidative damage. Our data provide clues for the development of control technology for agricultural and forestry pests as the silkworm is a representative of lepidopteran pests.
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