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Molecular cloning and characterization of cystatin, a cysteine protease inhibitor, from Angiostrongylus cantonensis

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Abstract

Cystatins are thiol proteinase inhibitors ubiquitously present in mammalian body and serve various important physiological functions. In the present study, a novel cystatin molecule (AcCystatin) was cloned from a cDNA library of Angiostrongylus cantonensis fourth-stage larvae. The putative 14-kDa protein contained 120 residues with cystatin-conserved motifs known to interact with the active site of cysteine peptidases and showed high identities with cystatins from other nematodes. RT-PCR analysis revealed that the expression pattern of AcCystatin was equal at the time points of third-stage larvae, fourth-stage larvae, and adults of the parasite life cycle. The recombinant AcCystatin (rAcCystatin) expressed and purified from Escherichia coli has been demonstrated to possess an obvious inhibitory activity against cathepsin B and could significantly upregulate nitric oxide production from IFN-γ activated RAW 264.7 macrophages. Sera from mice (non-permissive host) infected with A. cantonensis detected rAcCystatin by Western blot, while the sera from infected rats (permissive host) could not. The results implied that AcCystatin might be an immunoregulator in A. cantonensis infection.

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Acknowledgments

We are grateful to Pro. Qing-yu He, Mrs. Jie Liu, Dr. Jue-heng Wu, and Dr. Xun Zhu for their expert technical assistances in mass spectrometry, confocal microscopy, and fluorescence microplate reader, respectively. This work was supported by a grant from the National Basic Research Program of China (2010CB530004), the National Natural Science Foundation of China (30771888, 30800966), and Research Fund for Students of Sun Yat-sen University (2010).

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Authors and Affiliations

  1. Department of Parasitology, Zhongshan School of Medicine, Sun Yat-sen University, 74 2nd Zhongshan Road, Guangzhou, 510080, China

    Yu-hong Liu, Yan-ping Han, Zheng-yu Li, Jie Wei, Han-jiang He, Chang-zhi Xu, Huan-qin Zheng, Xi-mei Zhan, Zhong-dao Wu & Zhi-yue Lv

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

    Yu-hong Liu, Yan-ping Han, Zheng-yu Li, Jie Wei, Han-jiang He, Chang-zhi Xu, Huan-qin Zheng, Xi-mei Zhan, Zhong-dao Wu & Zhi-yue Lv

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  1. Yu-hong Liu
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  2. Yan-ping Han
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  3. Zheng-yu Li
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Correspondence to Zhong-dao Wu or Zhi-yue Lv.

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Yu-hong Liu and Yan-ping Han made equal contribution to this work.

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Liu, Yh., Han, Yp., Li, Zy. et al. Molecular cloning and characterization of cystatin, a cysteine protease inhibitor, from Angiostrongylus cantonensis . Parasitol Res 107, 915–922 (2010). https://doi.org/10.1007/s00436-010-1952-5

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  • DOI: https://doi.org/10.1007/s00436-010-1952-5

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