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Identification and characterization of a member of Rab subfamily, Rab8, from Clonorchis sinensis

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Abstract

The Rabs act as a binary molecular switch that utilizes the conformational changes associated with the GTP/GDP cycle to elicit responses from target proteins. It regulates a broad spectrum of cellular processes including cell proliferation, cytoskeletal assembly, and intracellular membrane trafficking in eukaryotes. The Rab8 from Clonorchis sinensis (CsRab8) was composed of 199 amino acids. The deduced amino acid sequence shared above 50 % identities with other species from trematode, tapeworm, mammal, insecta, nematode, and reptile, respectively. The homologous analysis of sequences showed the conservative domains: G1 box (GDSGVGKS), G2 box (T), G3 box (DTAG), G4 box (GNKCDL), and G5 box. In addition, the structure modeling had also shown other functional domains: GTP/Mg2+ binding sites, switch I region, and switch II region. A phylogenic tree analysis indicated that the CsRab8 was clustered with the Rab from Schistosoma japonicum, and trematode and tapeworm came from the same branch, which was different from an evolutional branch built by other species, such as mammal animal, insecta, nematode, and reptile. The recombinant CsRab8 protein was expressed in Escherichia coli and the purified protein was a soluble molecule by 12 % sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis. CsRab8 was identified as a component of excretory/secretory products of C. sinensis by western blot analysis. The transcriptional level of CsRab8 at metacercaria stage was the highest at the four stages and higher by 56.49-folds than that at adult worm, 1.23-folds than that at excysted metacercaria, and 2.69-folds than that at egg stage. Immunohistochemical localization analysis showed that CsRab8 was specifically distributed in the tegument, vitellarium, eggs, and testicle of adult worms, and detected on the vitellarium and tegument of metacercaria. Combined with the results, CsRab8 is indispensable for survival and development of parasites, especially for regulating excretory/secretory products secretion.

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Acknowledgments

This work was supported by National Key Basic Research and Development Project of China (973 project; No. 2010CB530000), National Natural Science Foundation of China (No. 81101270 and No. 81171602), National S & T Major Program (No. 2012ZX10004-220), and Fundamental Research Funds for the Central Universities (No. 3164035 and No. 3161036).

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Authors and Affiliations

  1. Department of Pathogen Biology, Hainan Medical College, Haikou, Hainan, 571199, China

    Pei Liang & Gang Lu

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

    Pei Liang, Jinyun Yu, Zhizhi Xie, Qiang Mao, Chi Liang, Yan Huang & Xinbing Yu

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

    Pei Liang, Jinyun Yu, Zhizhi Xie, Qiang Mao, Chi Liang, Yan Huang & Xinbing Yu

  4. Department of Clinical Laboratory, Guangzhou Red Cross Hospital, Medical College of Jinan University, Guangzhou, 510220, China

    Lei He

  5. Department of Clinical Laboratory, The First Affiliated Hospital of Guangzhou, University of Chinese Medicine, Guangzhou, 510080, China

    Xueqing Chen

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  1. Pei Liang
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  2. Lei He
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Correspondence to Gang Lu or Xinbing Yu.

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Liang, P., He, L., Yu, J. et al. Identification and characterization of a member of Rab subfamily, Rab8, from Clonorchis sinensis . Parasitol Res 114, 1857–1864 (2015). https://doi.org/10.1007/s00436-015-4372-8

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  • DOI: https://doi.org/10.1007/s00436-015-4372-8

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