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Molecular characterization and serological reactivity of a vacuolar ATP synthase subunit ε-like protein from Clonorchis sinensis

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Abstract

The vacuolar ATPase enzyme complex (V-ATPase) pumps protons across membranes, energized by hydrolysis of ATP. Extensive investigations on structural and biochemical features of these molecules have implied their importance in the physiological process. In this study, a full-length sequence encoding a vacuolar ATP synthase subunit ε-like protein of Clonorchis sinensis (CsATP-ε) was isolated from our cDNA library. The hypothetical 226 amino acid sequence shared 76 % identity with ATP-ε proteins of Schistosoma japonicum and above 55 % identity with ATP-ε proteins from human and other eukaryotes. Characteristic Asp140 amino acid residues and seven B-cell epitopes were predicted in this sequence. The complete coding sequence of the gene was expressed in Escherichia coli. Recombinant CsATP-ε (rCsATP-ε) protein could be probed by anti-rCsATP-ε rat serum and C.sinensis-infected human serum in Western blotting experiment, indicating that it is an antigen of strong antigenicity. The high level of antibody titers (1:204,800) showed that CsATP-ε has a powerful immunogenicity. Both the increased level and the change trend of IgG1/IgG2a subtypes in serum showed that the rCsATP-ε can induce strong combined Th1/Th2 immune responses in rats and stimulate the immune response changes to the dominant Th2 from Th1 along with long time infection. The results of immunoblot and immunolocalization demonstrated that CsATP-ε was consecutively expressed at various developmental stages of the parasite, which was supported by real-time PCR analysis. In immunohistochemistry, CsATP-ε was localized on the intestine, vitellarium, and testicle of an adult worm and excretory bladder of metacercaria, implying that CsATP-ε may relate to energy intake and metabolism. This fundamental study would contribute to further researches that are related to growth and development and immunomodulation of C. sinensis.

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Acknowledgment

This work was supported by the Development Program of China (973 program; No. 2010CB530000).

Conflict of interest

All authors declared that they have no competing interests.

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Authors and Affiliations

  1. Department of Medical Laboratory and Research Center, Tangdu Hospital, Fourth Military Medical University, Xi’an, Shannxi, 710038, China

    Xiaoli Lv

  2. Department of Clinical Laboratory, Sun Yat-sen Memorial Hospital of Sun Yat-sen University, Guangzhou, Guangdong, 510120, China

    Lisi Huang

  3. Department of Parasitology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, Guangdong, 510080, China

    Wenjun Chen, Xiaoyun Wang, Yan Huang, Chuanhuan Deng, Jiufeng Sun, Yanli Tian, Qiang Mao, Huali Lei & Xinbing Yu

  4. Key Laboratory for Tropical Diseases Control, Ministry of Education, Sun Yat-sen University, Guangzhou, Guangdong, 510080, China

    Wenjun Chen, Xiaoyun Wang, Yan Huang, Chuanhuan Deng, Jiufeng Sun, Yanli Tian, Qiang Mao, Huali Lei & Xinbing Yu

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  1. Xiaoli Lv
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  2. Lisi Huang
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  3. Wenjun Chen
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  11. Xinbing Yu
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Correspondence to Xinbing Yu.

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Xiaoli Lv and Lisi Huang contributed equally to this study

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Lv, X., Huang, L., Chen, W. et al. Molecular characterization and serological reactivity of a vacuolar ATP synthase subunit ε-like protein from Clonorchis sinensis . Parasitol Res 113, 1545–1554 (2014). https://doi.org/10.1007/s00436-014-3799-7

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