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Identification and molecular characterization of a novel signaling molecule 14-3-3 epsilon in Clonorchis sinensis excretory/secretory products

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Abstract

Increasing evidence shows that 14-3-3 proteins are involved in many biology events in addition to signal transduction. Extensive investigations on structural and biochemical features of these signaling molecules have implied their importance in the biological process. In the present study, we have identified and characterized the 14-3-3 epsilon (Cs14-3-3) in Clonorchis sinensis that causes human clonorchiasis. Recombinant protein was expressed in Escherichia coli (E. coli) and identified by MALDI-TOF/TOF. Immunoblot results revealed that Cs14-3-3 was a component of excretory/secretory products. Ligand blot assay indicated that 14-3-3 epsilon could bind C. sinensis MAPKAPK 2 in a nonphosphorylation-dependent manner. This protein could be detected at four stages of the life cycle by RT-PCR experiments and immunolocalization showed that Cs14-3-3 was extensively distributed in C. sinensis, especially at the outer surface and the sucker of adult worm and cyst wall of metacercaria. Taken together, 14-3-3 epsilon might play some roles in the development of the parasites. In addition, Cs14-3-3 epsilon should be addressed for the diagnostic value in C. sinensis infection in consideration of high sensitivity and specificity. As an immune stimulus, C. sinensis 14-3-3 epsilon was found to provoke a Th1/Th2 balanced immune response by inducing high levels of both IgG1 and IgG2a. Recombinant Cs14-3-3 conferred effective protection both in worm reduction rate and egg reduction rate, suggesting that the signaling molecule Cs14-3-3 was a promising vaccine candidate against C. sinensis infection.

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Acknowledgement

This work was supported by the Development Program of China (973 program; No. 2010CB530000) and the Sun Yat-sen University Innovative Talents Cultivation Program for Excellent Tutors.

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Authors and Affiliations

  1. Department of Parasitology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, 510080, People’s Republic of China

    Xiaoyun Wang, Wenjun Chen, Xuerong Li, Chenhui Zhou, Chuanhuan Deng, Xiaoli Lv, Yongxiu Fan, Jingtao Men, Chi Liang & Xinbing Yu

  2. Key Laboratory for Tropical Diseases Control, Sun Yat-sen University, Ministry of Education, Guangzhou, 510080, People’s Republic of China

    Xiaoyun Wang, Wenjun Chen, Xuerong Li, Chenhui Zhou, Chuanhuan Deng, Xiaoli Lv, Yongxiu Fan, Jingtao Men, Chi Liang & Xinbing Yu

  3. Department of Gynecology and Pediatrics, Nursing School, Guangdong Medical College, Dongguan, 523808, People’s Republic of China

    Chenhui Zhou

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  1. Xiaoyun Wang
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  2. Wenjun Chen
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  10. Xinbing Yu
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Correspondence to Xinbing Yu.

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Xiaoyun Wang and Wenjun Chen contributed equally to this work

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Wang, X., Chen, W., Li, X. et al. Identification and molecular characterization of a novel signaling molecule 14-3-3 epsilon in Clonorchis sinensis excretory/secretory products. Parasitol Res 110, 1411–1420 (2012). https://doi.org/10.1007/s00436-011-2642-7

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