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引用本文:刘 超,赵 培,梁 艳,高 强,刘升平,黄 倢.中国明对虾精氨酸激酶与白斑综合征病毒结构蛋白VP31的作用初探[J].海洋科学,2014,38(3):104-110.
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中国明对虾精氨酸激酶与白斑综合征病毒结构蛋白VP31的作用初探
刘 超1,2, 赵 培2, 梁 艳2, 高 强2, 刘升平1, 黄 倢2
1.青岛农业大学 动物科技学院;2.中国水产科学院 黄海水产研究所
摘要:
白斑综合征病毒(white spot syndrome virus, WSSV)是危害对虾的主要病原, 给全球水产养殖业带来了巨大经济损失。为研究阻断WSSV 的途径, 前期研究WSSV 与宿主相结合的受体蛋白发现, 白斑综合征病毒囊膜蛋白VP31 可以与中国明对虾(Fenneropenaeus chinensis)精氨酸激酶(FcAK)发生结合作用。精氨酸激酶(AK)通过调节无脊椎动物体内磷酸精氨酸与三磷酸腺苷(ATP)之间平衡在能量代谢、储存和利用方面具有重要作用。作者通过重组表达的rFcAK 与rVP31 的far-Western blotting 分析, 证实两者有结合作用, 此外, rFcAK 还与WSSV 的VP19、VP28 等6 种结构蛋白有结合作用。双向电泳分析观察到, rFcAK 与rVP31 的磷酸化反应后, rVP31 部分发生pI 降低现象, 提示rVP31 可能发生了磷酸化。生物信息学分析表明VP31 存在21 个精氨酸残基, FcAK 具有12 个精氨酸结合位点, 这可能为VP31和FcAK 的结合活性提供了靶位。为进一步研究WSSV 与宿主结合机理及阻断感染的机制提供思路。
关键词:  精氨酸激酶(AK)  白斑综合征病毒(WSSV)  中国明对虾(Fenneropenaeus chinensis)  结合
DOI:10.11759/hykx20120702003
分类号:
基金项目:现代农业产业技术体系专项资助项目(CARS-47); 公益性行业(农业)科研专项经费资助项目(201103034)
Preliminary study on the conjunction between arginine kinase of Fenneropenaeus chinensis and the structure proteins of white spot syndrome virus (WSSV)
Abstract:
White spot syndrome virus (WSSV) is a major shrimp pathogen causing big economic losses all over the world. In order ro explore ways to block WSSV infection, we worked on WSSV-binding host receptor proteins, and we found that envelope protein VP31 of WSSV was able to engage with arginine kinase of Fenneropenaeus chinensis (FcAK). Arginine kinase (AK) plays important roles in the metabolism, storage and utilization of energy by regulation of the balance between phosphorylation of arginine and ATP in invertebrate. The present study proves the binding between the recombinant FcAK (rFcAK) and VP31 (rVP31) using far-Western blotting. Furthermore, the far-Western blotting also showed that rFcAK was able to bind other 6 structural proteins of WSSV, including VP19 and VP28. Two-dimensional electrophoresis analysis showed that part of VP31 migrated to the spot with reduced pI after the in vitro phosphorylating incubation between rFcAK and rVP31, suggesting that the rVP31 might be phosphorylated. The bioinformatics analysis showed that VP31 had 21 arginine residents and FcAK had 12 arginine binding sites. They may provide the loci for the binding activity between VP31 and FcAK. We’ll further elucidate the underlying binding mechanism between WSSV and host, and try to find ways to block the infection of WSSV.
Key words:  Arginine kinase (AK)  WSSV  Fenneropenaeus chinensis  binding activity
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