Molecular characterization of cathepsin B from Clonorchis sinensis excretory/secretory products and assessment of its potential for serodiagnosis of clonorchiasis.
Molecular characterization of cathepsin B from Clonorchis sinensis excretory/secretory products and assessment of its potential for serodiagnosis of clonorchiasis.
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DOI：
10.1186/1756-3305-4-149
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发表时间：
2011-07-27
影响因子：
3.2
通讯作者：
Yu X
中科院分区：
文献类型：
--
作者：
Chen W;Wang X;Li X;Lv X;Zhou C;Deng C;Lei H;Men J;Fan Y;Liang C;Yu X
组织蛋白酶半胱氨酸蛋白酶在寄生虫的生命周期中发挥多种作用，例如食物摄取、免疫入侵和发病机制，使其成为诊断分析、疫苗和药物的有价值的靶标。本研究的目的是鉴定华支睾吸虫（CsCB）的组织蛋白酶 B，并探讨其对人类蠕虫病的诊断价值。中华线虫组织蛋白酶B的预测氨基酸序列与日本血吸虫、智人和肝片形吸虫组织蛋白酶B的氨基酸序列同源性分别为63%、52%、50%。编码 CsCB 酶原的序列在大肠杆菌中过表达。逆转录PCR实验表明，CsCB在中华线虫成虫和囊蚴中都有转录。通过免疫印迹测定，CsCB被鉴定为中华虫的排泄/分泌产物，这与免疫组化定位一致，表明CsCB在中华虫成虫的肠道中特别表达。 ELISA 和蛋白质印迹分析均表明重组 CsCB 可以与华支睾吸虫病和其他蠕虫病的人血清发生反应。我们的研究结果表明，分泌的 CsCB 可能在中华蠕虫的生物学中发挥重要作用，并且可能成为蠕虫病的诊断候选者。
Cathepsin cysteine proteases play multiple roles in the life cycle of parasites such as food uptake, immune invasion and pathogenesis, making them valuable targets for diagnostic assays, vaccines and drugs. The purpose of this study was to identify a cathepsin B of Clonorchis sinensis (CsCB) and to investigate its diagnostic value for human helminthiases. The predicted amino acid sequence of the cathepsin B of C. sinensis shared 63%, 52%, 50% identity with that of Schistosoma japonicum, Homo sapiens and Fasciola hepatica, respectively. Sequence encoding proenzyme of CsCB was overexpressed in Escherichia coli. Reverse transcription PCR experiments revealed that CsCB transcribed in both adult worm and metacercaria of C. sinensis. CsCB was identified as a C. sinensis excretory/secretory product by immunoblot assay, which was consistent with immunohistochemical localization showing that CsCB was especially expressed in the intestine of C. sinensis adults. Both ELISA and western blotting analysis showed recombinant CsCB could react with human sera from clonorchiasis and other helminthiases. Our findings revealed that secreted CsCB may play an important role in the biology of C. sinensis and could be a diagnostic candidate for helminthiases.
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影响因子：
1.5
作者：
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通讯作者：
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