Abstract
Cathepsin L of parasite plays multiple roles in growth, food uptake, and invasion into host and pathogenesis, which makes it a valuable target for diagnosis, vaccine, and drug. In this study, we identified a cDNA encoding cathepsin L homolog (CsCPL) from the library of Clonorchis sinensis adult by bioinformatics analysis. Sequence encoding proenzyme of CsCPL (removal of signal peptide, CsproCPL) was highly expressed in form of inclusion body in Escherichia coli, and soluble rCsproCPL (about 1 mg/ml) in high purity were obtained after denaturation, purification, and renaturation. Western blot analysis indicated that CsCPL is a component of excretory–secretory products of adult, in mature form of protease. Reverse transcription polymerase chain reaction showed that CsCPL is also expressed in metacercaria and cercaria stage. Immunolocalization demonstrated that CsCPL is deposited at adult intestine, or tegument, and tegumentary cell of metacercaria and cercaria (especially at dorsal tegument of cercaria), indicating different secretory routine roles in adult and larva. The characteristics of CsCPL suggested that it may involve in invasion of cercaria into fish and development to metacercaria, excystment of metacercaria, and protein digestion of adult, which may render it a candidate antigen for fish vaccine and serodiagnosis of human clonorchiasis.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Berasain P, Goni F, McGonigle S, Dowd A, Dalton JP, Frangione B, Carmona C (1997) Proteinases secreted by Fasciola hepatica degrade extracellular matrix and basement membrane components. J Parasitol 83:1–5
Berasain P, Carmona C, Frangione B, Dalton JP, Goni F (2000) Fasciola hepatica: parasite-secreted proteinases degrade all human IgG subclasses: determination of the specific cleavage sites and identification of the immunoglobulin fragments produced. Exp Parasitol 94(2):99–110
Carmona C, Dowd AJ, Smith AM, Dalton JP (1993) Cathepsin L proteinase secreted by Fasciola hepatica in vitro prevents antibody-mediated eosinophil attachment to newly excysted juveniles. Mol Biochem Parasitol 62(1):9–17
Chappell CL, Dresden MH (1986) Schistosoma mansoni: proteinase activity of “hemoglobinase” from the digestive tract of adult worms. Exp Parasitol 61:160–167
Choi BI, Han JK, Hong ST, Lee KH (2004) Clonorchiasis and cholangiocarcinoma: etiologic relationship and imaging diagnosis. Clin Microbiol Rev 17(3):540–552
Dalton JP, Neill SO, Stack C, Collins P, Walshe A, Sekiya M, Doyle S, Mulcahy G, Hoyle D, Khaznadji E, Moiré N, Brennan G, Mousley A, Kreshchenko N, Maule AG, Donnelly SM (2003) Fasciola hepatica cathepsin L-like proteases:biology, function, and potential in the development of first generation liver fluke vaccines. Int J Parasitol 33(11):1173–1181
Guiliano DB, Hong X, McKerrow JH, Blaxter ML, Oksov Y, Liu J, Ghedin E, Lustiqman S (2004) A gene family of cathepsin L-like proteases of filarial nematodes are associated with larval molting and cuticle and eggshell remodeling. Mol Biochem Parasitol 136(2):227–242
Hu X, Li Y, Xu J, Hu F, Zhao J, Yu X (2008) Cloning and bioinformatic analysis of cathepsin L1-like full-length gene from Clonorchis sinensis. J Pathogen Biology 3(7):508–513
Kim J, Chai JY, Kho WG, Cho KH, Lee SH (1991) Immunohistochemical study on the antigenicity of each organ structure of Clonorchis sinensis. Korean J Parasitol 29(1):21–29
Lim MK, Ju YH, Franceschi S, Oh JK, Kong HJ, Hwang SS, Park SK, Cho SI, Sohn WM, Kim DI, Yoo KY, Hong ST, Shin HR (2006) Clonorchis sinensis infection and increasing risk of cholangiocarcinoma in the Republic of Korea. Am J Trop Med Hyg 75(1):93–96
Lun ZR, Gasser RB, Lai DH, Li AX, Zhu XQ, Yu XB, Fang YY (2005) Clonorchiasis: a key foodborne zoonosis in China. Lancet Infect Dis 5:31–41
McKerrow JH, Doenhoff MJ (1988) Schistosome proteases. Parasitol Today 4(12):334–340
Nagano I, Pei F, Wu Z, Wu J, Cui H, Boonmars T, Takahashi Y (2004) Molecular expression of a cysteine proteinase of Clonorchis sinensis and its application to an enzyme-linked immunosorbent assay for immunodiagnosis of clonorchiasis. Clin Diagn Lab Immunol 11(2):411–416
Pan B, Fang YY, Yang WS (2000) Current situation and control strategy of parasitic diseases in Guangdong province. Ann Bull Soc Parasitol Guangdong 22:85–89 (in Chinese)
Sajid M, McKerrow JH (2002) Cysteine proteases of parasitic organisms. Mol Biochem Parasitol 120(1):1–21
Tort J, Brindley PJ, Knox D, Wolfe KH, Dalton JP (1999) Proteinases and associated genes of parasitic helminths. Adv Parasitol 43:161–266
Wongkham C, Tantrawatpan C, Intapan PM, Maleewong W, Wongkham S, Nakashima K (2005) Evaluation of Immunoglobulin G Subclass Antibodies against Recombinant Fasciola gigantica Cathepsin L1 in an Enzyme-Linked Immunosorbent Assay for Serodiagnosis of Human Fasciolosis. Clin Diagn Lab Immunol 12(10):1152–1156
Xu J, Hu X, Kang Y, Wu Z, Chen S, **e Y, Yu X (2004) Construction of full-length gene expression library of Clonorchis sinensis adults and establishment of the gene expression pattern. Chinese Journal of Zoonoses 20(5):383–393
Zhou Z, **a H, Hu X, Huang Y, Li Y, Li L, Ma C, Chen X, Hu F, Xu J, Lu F, Wu Z, Yu X (2008) Oral administration of a Bacillus subtilis spore-based vaccine expressing Clonorchis sinensis tegumental protein 22.3 kDa confers protection against Clonorchis sinensis. Vaccine 26(15):1817–1825
Acknowledgements
This work was supported by grants from the Guangzhou Key Problem Tackling Science-Technology Plan (No.2006Z3-E4011) and the National High Technology Research and Development Program of China (863 program; No.2006AA02Z422).
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Li, Y., Hu, X., Liu, X. et al. Molecular cloning and analysis of stage and tissue-specific expression of Cathepsin L-like protease from Clonorchis sinensis . Parasitol Res 105, 447–452 (2009). https://doi.org/10.1007/s00436-009-1406-0
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00436-009-1406-0