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Recombinant adenylate kinase 3 from liver fluke Clonorchis sinensis for histochemical analysis and serodiagnosis of clonorchiasis

  • Soon Bin Kwon (a1) (a2), Paul Kim (a1) (a2), Hae Sun Woo (a3), Tae Yun Kim (a4), Ju Yeong Kim (a3), Hye Min Lee (a1) (a2), Yun Soo Jang (a5), Eun-Min Kim (a3), Tai-Soon Yong (a3) and Baik Lin Seong (a1) (a2)...


Due to the lack of an effective prophylactic intervention and diagnosis, human liver fluke Clonorchis sinensis continues to afflict a large human population, causing a chronic inflammatory bile duct disease. With an aim to identify target antigens for sensitive serodiagnosis, adenylate kinase 3 of C. sinensis (CsAK3) was successfully expressed in soluble form in Escherichia coli by fusion to an RNA-interacting domain derived from human Lys-tRNA synthetase and purified by Ni2+-affinity chromatography. Anti-CsAK3 serum was raised by immunization of mice, and Western blotting confirmed that CsAK3 was expressed in adult-stage C. sinensis. Histochemical analysis showed that CsAK3 was localized to the subtegumental tissue of C. sinensis and was excreted into the bile duct of the host. When tested against sera from various parasite-infected patients by enzyme-linked immunosorbent assay, the recombinant CsAK3 elicited a specific response to C. sinensis-infected sera. The results suggest that CsAK3, either alone or in combination with other antigens, could be used for improving the clinical diagnosis of clonorchiasis.


Corresponding author

Author for correspondence: Baik Lin Seong, E-mail:


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Attwood, HD and Chou, ST (1978) The longevity of Clonorchis sinensis. Pathology 10, 153156.
Bian, M, Li, S, Wang, X, Xu, Y, Chen, W, Zhou, C, Chen, X, He, L, Xu, J, Liang, C, Wu, Z, Huang, Y, Li, X and Yu, X (2014) Identification, immunolocalization, and immunological characterization of nitric oxide synthase-interacting protein from Clonorchis sinensis. Parasitology Research 113, 17491757.
Cho, S, Chung, B and Lee, S (2013) National survey of intestinal parasitic infections in Korea, 8th Report 2013. 2005-811X. Korea centers for disease control & prevention, Seoul, Korea.
Choi, D, Lim, JH, Lee, KT, Lee, JK, Choi, SH, Heo, JS, Jang, KT, Lee, NY, Kim, S and Hong, ST (2006) Cholangiocarcinoma and Clonorchis sinensis infection: a case-control study in Korea. Journal of Hepatology 44, 10661073.
Choi, MH, Park, IC, Li, S and Hong, ST (2003) Excretory-secretory antigen is better than crude antigen for the serodiagnosis of clonorchiasis by ELISA. The Korean Journal of Parasitology 41, 3539.
Choi, SI, Han, KS, Kim, CW, Ryu, KS, Kim, BH, Kim, KH, Kim, SI, Kang, TH, Shin, HC, Lim, KH, Kim, HK, Hyun, JM and Seong, BL (2008) Protein solubility and folding enhancement by interaction with RNA. PLoS ONE 3, e2677.
Choi, SI, Ryu, K and Seong, BL (2009) RNA-mediated chaperone type for de novo protein folding. RNA Biology 6, 2124.
Choi, SI, Kwon, S, Son, A, Jeong, H, Kim, KH and Seong, BL (2013) Protein folding in vivo revisited. Current Protein and Peptdie Science 14, 721733.
Choi, SI, Lim, KH and Seong, BL (2011) Chaperoning roles of macromolecules interacting with proteins in vivo. International Journal of Molecular Sciences 12, 19791990.
Choi, SI, Son, A, Lim, KH, Jeong, H and Seong, BL (2012) Macromolecule-assisted de novo protein folding. International Journal of Molecular Sciences 13, 1036810386.
Dereeper, A, Guignon, V, Blanc, G, Audic, S, Buffet, S, Chevenet, F, Dufayard, JF, Guindon, S, Lefort, V, Lescot, M, Claverie, JM and Gascuel, O (2008) robust phylogenetic analysis for the non-specialist. Nucleic Acids Research 36, W465W469.
Dereeper, A, Audic, S, Claverie, JM and Blanc, G (2010) BLAST-EXPLORER helps you building datasets for phylogenetic analysis. BMC Evolutionary Biology 10, 8.
Desogus, G, Todone, F, Brick, P and Onesti, S (2000) Active site of lysyl-tRNA synthetase: structural studies of the adenylation reaction. Biochemistry 39, 84188425.
Docter, BE, Horowitz, S, Gray, MJ, Jakob, U and Bardwell, JC (2016) Do nucleic acids moonlight as molecular chaperones? Nucleic Acids Research 44, 48354845.
Francin, M, Kaminska, M, Kerjan, P and Mirande, M (2002) The N-terminal domain of mammalian Lysyl-tRNA synthetase is a functional tRNA-binding domain. Journal of Biological Chemistry 277, 17621769.
Hong, ST and Fang, Y (2012) Clonorchis sinensis and clonorchiasis, an update. Parasitology International 61, 1724.
Huang, L, Lv, X, Huang, Y, Hu, Y, Yan, H, Zheng, M, Zeng, H, Li, X, Liang, C, Wu, Z and Yu, X (2014) Identification, sequence analysis, and characterization of serine/threonine protein kinase 17A from Clonorchis sinensis. Parasitology Research 113, 17131723.
Huang, Y, Zheng, Y, Li, Y, Yang, M, Li, T, Zeng, S, Yu, X, Huang, H and Hu, X (2013) Expression, immunolocalization, and serological reactivity of a novel sphingomyelin phosphodiesterase-like protein, an excretory/secretory antigen from Clonorchis sinensis. Parasitology Research 112, 21972206.
Jang, YH, Cho, SH, Son, A, Lee, YH, Lee, J, Lee, KH and Seong, BL (2014) High-yield soluble expression of recombinant influenza virus antigens from Escherichia coli and their potential uses in diagnosis. Journal of Virological Methods 196, 5664.
Ju, JW, Joo, HN, Lee, MR, Cho, SH, Cheun, HI, Kim, JY, Lee, YH, Lee, KJ, Sohn, WM, Kim, DM, Kim, IC, Park, BC and Kim, TS (2009) Identification of a serodiagnostic antigen, legumain, by immunoproteomic analysis of excretory-secretory products of Clonorchis sinensis adult worms. Proteomics 9, 30663078.
Kall, L, Krogh, A and Sonnhammer, EL (2004) A combined transmembrane topology and signal peptide prediction method. Journal of Molecular Biology 338, 10271036.
Kall, L, Krogh, A and Sonnhammer, EL (2005) An HMM posterior decoder for sequence feature prediction that includes homology information. Bioinformatics (oxford, England) 21(suppl. 1), i251i257.
Kall, L, Krogh, A and Sonnhammer, EL (2007) Advantages of combined transmembrane topology and signal peptide prediction – the Phobius web server. Nucleic Acids Research 35, W429W432.
Kim, HK, Cheun, HI, Cheun, BS, Lee, KY, Kim, TS, Lee, SE, Lee, WJ and Cho, SH (2010a) Prevalence of Clonorchis sinensis infections along the five major rivers in Republic of Korea, 2007. Osong Public Health and Research Perspectives 1, 4349.
Kim, HK, Choi, SI and Seong, BL (2010b) 5S rRNA-assisted DnaK refolding. Biochemical and Biophysical Research Communications 391, 11771181.
Kim, JM, Choi, HS and Seong, BL (2017) The folding competence of HIV-1 Tat mediated by interaction with TAR RNA. RNA Biology 14, 926937.
Kim, SI (1998) A Clonorchis sinensis-specific antigen that detects active human clonorchiasis. The Korean Journal of Parasitology 36, 3745.
Lee, JH, Rim, HJ and Bak, UB (1993) Effect of Clonorchis sinensis infection and dimethylnitrosamine administration on the induction of cholangiocarcinoma in Syrian golden hamsters. The Korean Journal of Parasitology 31, 2130.
Li, S, Chung, BS, Choi, MH and Hong, ST (2004) Organ-specific antigens of Clonorchis sinensis. The Korean Journal of Parasitology 42, 169174.
Li, S, Shin, JG, Cho, PY, Kim, TI, Hong, ST and Hong, SJ (2011) Multiple recombinant antigens of Clonorchis sinensis for serodiagnosis of human clonorchiasis. Parasitology Research 108, 12951302.
Li, Y, Hu, X, Liu, X, Huang, Y, Xu, J, Zhao, J, Wu, Z and Yu, X (2012) Serological diagnosis of clonorchiasis: using a recombinant propeptide of cathepsin L proteinase from Clonorchis sinensis as a candidate antigen. Parasitology Research 110, 21972203.
Liang, P, Sun, J, Huang, Y, Zhang, F, Zhou, J, Hu, Y, Wang, X, Liang, C, Zheng, M, Xu, Y, Mao, Q, Hu, X, Li, X, Xu, J, Lu, G and Yu, X (2013a) Biochemical characterization and functional analysis of fructose-1,6-bisphosphatase from Clonorchis sinensis. Molecular Biology Reports 40, 43714382.
Liang, P, Zhang, F, Chen, W, Hu, X, Huang, Y, Li, S, Ren, M, He, L, Li, R, Li, X, Xu, J, Wu, Z, Lu, G and Yu, X (2013b) Identification and biochemical characterization of adenylate kinase 1 from Clonorchis sinensis. Parasitology Research 112, 17191727.
Loscher, T, Nothdurft, HD, Prufer, L, von Sonnenburg, F and Lang, W (1981) Praziquantel in clonorchiasis and opisthorchiasis. Tropenmedizin und Parasitologie 32, 234236.
Papachristou, GI, Schoedel, KE, Ramanathan, R and Rabinovitz, M (2005) Clonorchis sinensis-associated cholangiocarcinoma: a case report and review of the literature. Digestive Diseases and Sciences 50, 21592162.
Qian, MB, Chen, YD, Liang, S, Yang, GJ and Zhou, XN (2012) The global epidemiology of clonorchiasis and its relation with cholangiocarcinoma. Infectious Diseases of Poverty 1, 4.
Qing, G, Xia, B and Inouye, M (2003) Enhancement of translation initiation by A/T-rich sequences downstream of the initiation codon in Escherichia coli. Journal of Molecular Microbiology and Biotechnology 6, 133144.
Sim, S, Park, GM and Yong, TS (2003) Cloning and characterization of Clonorchis sinensis myoglobin using immune sera against excretory-secretory antigens. Parasitology Research 91, 338343.
Son, A, Choi, SI, Han, G and Seong, BL (2015) M1 RNA is important for the in-cell solubility of its cognate C5 protein: implications for RNA-mediated protein folding. RNA Biology 12, 11981208.
Tompa, P (2002) Intrinsically unstructured proteins. Trends in Biochemical Sciences 27, 527533.
Wang, X, Chen, W, Tian, Y, Mao, Q, Lv, X, Shang, M, Li, X, Yu, X and Huang, Y (2014) Surface display of Clonorchis sinensis enolase on Bacillus subtilis spores potentializes an oral vaccine candidate. Vaccine 32, 13381345.
Watanapa, P and Watanapa, WB (2002) Liver fluke-associated cholangiocarcinoma. The British Journal of Surgery 89, 962970.
Yang, G, Yu, X, Wu, Z, Xu, J, Song, L, Zhang, H, Hu, X, Zheng, N, Guo, L, Xu, J, Dai, J, Ji, C, Gu, S and Ying, K (2005) Molecular cloning and characterization of a novel adenylate kinase 3 gene from Clonorchis sinensis. Parasitology Research 95, 406412.
Yiadom, KP, Hammamieh, R, Ukpabi, N, Tsang, P and Yang, DC (2003) A peptide from the extension of Lys-tRNA synthetase binds to transfer RNA and DNA. Peptides 24, 987998.
Yoo, WG, Kim, DW, Ju, JW, Cho, PY, Kim, TI, Cho, SH, Choi, SH, Park, HS, Kim, TS and Hong, SJ (2011) Developmental transcriptomic features of the carcinogenic liver fluke, Clonorchis sinensis. PLoS Neglected Tropical Diseases 5, e1208.
Young, ND, Nagarajan, N, Lin, SJ, Korhonen, PK, Jex, AR, Hall, RS, Safavi-Hemami, H, Kaewkong, W, Bertrand, D, Gao, S, Seet, Q, Wongkham, S, Teh, BT, Wongkham, C, Intapan, PM, Maleewong, W, Yang, X, Hu, M, Wang, Z, Hofmann, A, Sternberg, PW, Tan, P, Wang, J and Gasser, RB (2014) The Opisthorchis viverrini genome provides insights into life in the bile duct. Nature Communications 5, 4378.
Zheng, S, Zhu, Y, Zhao, Z, Wu, Z, Okanurak, K and Lv, Z (2017) Liver fluke infection and cholangiocarcinoma: a review. Parasitology Research 116, 1119.
Zhou, C, Bian, M, Liao, H, Mao, Q, Li, R, Zhou, J, Wang, X, Li, S, Liang, C, Li, X, Huang, Y and Yu, X (2013) Identification and immunological characterization of thioredoxin transmembrane-related protein from Clonorchis sinensis. Parasitology Research 112, 17291736.
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