数字农科院2.0

Characterization of aspartyl aminopeptidase from Schistosoma japonicum

文献类型： 外文期刊

作者： Zheng Shang;Qinghong Guo;Xue Zhou;Yongcheng Yue;Kerou Zhou;Liying Tang;Zhizhong Zhang;Zhiqiang Fu;Jinming Liu;Jiaojiao Lin;Bin Xu;Min Zhang;Yang Hong

作者机构：

关键词： Aspartyl aminopeptidase;Enzymatic activity;Immunoprotection;Schistosoma japonicum;Tegument

期刊名称： Acta Tropica

ISSN： 0001-706X

年卷期： 2022 年 232 卷

页码：

收录情况： SCIE（2022版）

摘要： The tegument of schistosomes is the interface between the worm and the host environment. Some molecules distributed on the tegument participate in host–parasite interactions. Aspartyl aminopeptidase (AAP), identified on the tegument of Schistosoma japonicum (S. japonicum), facilitate protein turnover by acting in concert with other aminopeptidases. In this study, the gene encoding S. japonicum aspartyl aminopeptidase (SjAAP) was cloned, expressed and characterized. Quantitative real-time PCR analysis showed that SjAAP was expressed in all studied developmental stages. The transcript level was higher in 8, 14, 21, and 28 days old worms than the other detected stages. Moreover, the level of expression in 42-day-old male worms was significantly higher than that in females. The recombinant SjAAP (rSjAAP) was expressed as both supernatant and inclusion bodies in Escherichia coli BL21 cells. The enzymatic activity of rSjAAP was 4.45 U/mg. The Km and Vmax values for H-Asp-pNA hydrolysis were discovered to be 5.93 mM and 0.018 mM·min–1. Immunofluorescence analysis revealed that SjAAP is primarily distributed on the tegument and parenchyma of schistosomes. Western blot showed that rSjAAP possessed good immunogenicity. Although specific antibodies were produced in BALB/c mice vaccinated with rSjAAP emulsified with ISA 206 adjuvant, no significant reduction of worm burden and number of eggs in the liver was observed. Therefore, rSjAAP may not be suitable to act as a potential vaccine candidate against schistosomiasis japonica in mice. However, this study provides some foundation for further exploration of the biological function of this molecule.

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