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Proteomic analysis of excretory secretory products from Clonorchis sinensis adult worms: molecular characterization and serological reactivity of a excretory–secretory antigen-fructose-1,6-bisphosphatase

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Abstract

Clonorchis sinensis is a food-borne zoonotic parasite that resides in bile ducts and causes clonorchiasis, which may result in cholelithiasis, cholecystitis, hepatic fibrosis, and liver tumors. Although total excretory secretory products (ESP) of C. sinensis adults induce hepatic fibrosis in vivo in rats, the causative mechanism is not well understood. To study components of the ESP, C. sinensis culture medium was collected and analyzed using shotgun LC–MS/MS. We identified a total of 110 proteins, including glycometabolic enzymes (such as fructose-1,6-bisphosphatase (FBPase) and enolase), detoxification enzymes (such as glutamate dehydrogenase, dihydrolipoamide dehydrogenase and cathepsin B endopeptidase), and a number of RAB family proteins. To identify a potential causative agent for hepatic fibrosis, we expressed and purified a recombinant FBPase, a 1,041-bp gene product that encodes a 41.7-kDa protein with prototypical FBPase domains and that can form a tetramer with a molecular mass of 166.8 kDa. In addition, we found that FBPase is an antigen present in the ESP and in circulation. Immunofluorescence showed that FBPase localizes to the intestinal cecum and vitellarium in C. sinensis adults. Our results describe the components of the excretory secretory products from C. sinensis adult worms and suggest that FBPase may be an important antigen present in the ESP of C. sinensis and may lay the foundation for additional studies on the development of clonorchiasis-associated hepatic fibrosis.

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References

  • Chen X, Hu X, Wu Z, Yu X, Ma C, Zhou Z (2007) Immunological cross-reactivity analysis on recombinant histamine-releasing factors from Schistosoma japonicum, Clonorchis sinensis, and Wistar rat. Parasitol Res 100:749–754

    Article  PubMed  Google Scholar 

  • Ch’In KY, Lei AT, Wang TY (1955) Primary mucinous carcinoma of liver associated with Clonorchis sinensis infection. Chin Med J 73:26–35

    PubMed  Google Scholar 

  • Chung YY, Ko RC (1999) A novel cDNA clone encoding a specific excretory/secretory antigen of larval Trichinella pseudospiralis. Parasitol Res 85:685–691

    Article  PubMed  CAS  Google Scholar 

  • Donahue JL, Bownas JL, Niehaus WG, Larson TJ (2000) Purification and characterization of glpX-encoded fructose 1, 6-bisphosphatase, a new enzyme of the glycerol 3-phosphate regulon of Escherichia coli. J Bacteriol 182:5624–5627

    Article  PubMed  CAS  Google Scholar 

  • Enes P, Panserat S, Kaushik S, Oliva-Teles A (2009) Nutritional regulation of hepatic glucose metabolism in fish. Fish Physiol Biochem 35:519–539

    Article  PubMed  CAS  Google Scholar 

  • Fried B, Abruzzi A (2010) Food-borne trematode infections of humans in the United States of America. Parasitol Res 106:1263–1280

    Article  PubMed  Google Scholar 

  • Gould TD, Quiroz JA, Singh J, Zarate CA, Manji HK (2004) Emerging experimental therapeutics for bipolar disorder: insights from the molecular and cellular actions of current mood stabilizers. Mol Psychiatry 9:734–755

    Article  PubMed  CAS  Google Scholar 

  • Gressner AM, Weiskirchen R (2006) Modern pathogenetic concepts of liver fibrosis suggest stellate cells and TGF-beta as major players and therapeutic targets. J Cell Mol Med 10:76–99

    Article  PubMed  CAS  Google Scholar 

  • Haas W, Faherty BK, Gerber SA, Elias JE, Beausoleil SA, Bakalarski CE, Li X, Villen J, Gygi SP (2006) Optimization and use of peptide mass measurement accuracy in shotgun proteomics. Mol Cell Proteomics 5:1326–1337

    Article  PubMed  CAS  Google Scholar 

  • Hu F, Chen W, Li L, Lu Y, Song W, Kuang Y, Zhang F (2009a) Cloning, characterization, and expression of a novel secretory lipase-like gene from Clonorchis sinensis. Parasitol Res 105:1661–1666

    Article  PubMed  Google Scholar 

  • Hu F, Hu X, Ma C, Zhao J, Xu J, Yu X (2009b) Molecular characterization of a novel Clonorchis sinensis secretory phospholipase A(2) and investigation of its potential contribution to hepatic fibrosis. Mol Biochem Parasitol 167:127–134

    Article  PubMed  CAS  Google Scholar 

  • Hu FY, Hu XC, Ma CL, Xu J, Yu XB (2009c) Excretory/secretory antigens from Clonorchis sinensis induces hepatic fibrosis in rats. Nan Fang Yi Ke Da Xue Xue Bao 29:393–396

    PubMed  CAS  Google Scholar 

  • Kepka A, Dariusz Szajda S, Stypulkowska A, Waszkiewicz N, Jankowska A, Chojnowska S, Zwierz K (2008) Urinary fructose-1,6-bisphosphatase activity as a marker of the damage to the renal proximal tubules in children with idiopathic nephrotic syndrome. Clin Chem Lab Med 46:831–835

    Article  PubMed  CAS  Google Scholar 

  • Kim YJ, Choi MH, Hong ST, Bae YM (2008) Proliferative effects of excretory/secretory products from Clonorchis sinensis on the human epithelial cell line HEK293 via regulation of the transcription factor E2F1. Parasitol Res 102:411–417

    Article  PubMed  Google Scholar 

  • Kim DM, Ko BS, Ju JW, Cho SH, Yang SJ, Yeom YI, Kim TS, Won Y, Kim IC (2009a) Gene expression profiling in mouse liver infected with Clonorchis sinensis metacercariae. Parasitol Res 106:269–278

    Article  PubMed  Google Scholar 

  • Kim YJ, Choi MH, Hong ST, Bae YM (2009b) Resistance of cholangiocarcinoma cells to parthenolide-induced apoptosis by the excretory-secretory products of Clonorchis sinensis. Parasitol Res 104:1011–1016

    Article  PubMed  Google Scholar 

  • Kuznetsova E, Xu L, Singer A, Brown G, Dong A, Flick R, Cui H, Cuff M, Joachimiak A, Savchenko A, Yakunin AF (2010) Structure and activity of the metal-independent fructose-1,6-bisphosphatase YK23 from Saccharomyces cerevisiae. J Biol Chem 285:21049–21059

    Article  PubMed  CAS  Google Scholar 

  • Lawton DE, Wigger H, Simcock DC, Simpson HV (2002) Effect of Ostertagia circumcincta excretory/secretory products on gastrin release in vitro. Vet Parasitol 104:243–255

    Article  PubMed  CAS  Google Scholar 

  • Lee JH, Rim HJ, Bak UB (1993) Effect of Clonorchis sinensis infection and dimethylnitrosamine administration on the induction of cholangiocarcinoma in Syrian golden hamsters. Korean J Parasitol 31:21–30

    Article  PubMed  CAS  Google Scholar 

  • Li S, Chung YB, Chung BS, Choi MH, Yu JR, Hong ST (2004) The involvement of the cysteine proteases of Clonorchis sinensis metacercariae in excystment. Parasitol Res 93:36–40

    Article  PubMed  Google Scholar 

  • 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

    Article  PubMed  Google Scholar 

  • Ma C, Hu X, Hu F, Li Y, Chen X, Zhou Z, Lu F, Xu J, Wu Z, Yu X (2007) Molecular characterization and serodiagnosis analysis of a novel lysophospholipase from Clonorchis sinensis. Parasitol Res 101:419–425

    Article  PubMed  Google Scholar 

  • Ma CL, Hu XC, Hu FY, Zhou HJ, Xue L, Yu XB (2008) The effects of Lysophospholipase from Clonorchis sinensis on the hepatic stellate cells and oval cells of rat. Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi 24:692–695

    PubMed  CAS  Google Scholar 

  • Nagano I, Wu Z, Takahashi Y (2009) Functional genes and proteins of Trichinella spp. Parasitol Res 104:197–207

    Article  PubMed  Google Scholar 

  • Pak JH, Kim DW, Moon JH, Nam JH, Kim JH, Ju JW, Kim TS, Seo SB (2009) Differential gene expression profiling in human cholangiocarcinoma cells treated with Clonorchis sinensis excretory-secretory products. Parasitol Res 104:1035–1046

    Article  PubMed  Google Scholar 

  • Rana SS, Bhasin DK, Nanda M, Singh K (2007) Parasitic infestations of the biliary tract. Curr Gastroenterol Rep 9:156–164

    Article  PubMed  Google Scholar 

  • Santra A, Chowdhury A, Ghosh A, Mazumder DN (2000) Development of an animal model of hepatic fibrosis by excretory-secretory antigen of Ascaris suum. Indian J Gastroenterol 19:119–121

    PubMed  CAS  Google Scholar 

  • Snabel V, Hanzelova V, Scholz T, Gerdeaux D, Cabaret J (2004) Allozyme analysis of genetic variation and polymorphism in Eubothrium salvelini and E. crassum (Cestoda: Pseudophyllidea) from alpine lakes. Parasitol Res 93:290–295

    Article  PubMed  Google Scholar 

  • Stec B, Yang H, Johnson KA, Chen L, Roberts MF (2000) MJ0109 is an enzyme that is both an inositol monophosphatase and the ‘missing’ archaeal fructose-1,6-bisphosphatase. Nat Struct Biol 7:1046–1050

    Article  PubMed  CAS  Google Scholar 

  • Strand A, Zrenner R, Trevanion S, Stitt M, Gustafsson P, Gardestrom P (2000) Decreased expression of two key enzymes in the sucrose biosynthesis pathway, cytosolic fructose-1,6-bisphosphatase and sucrose phosphate synthase, has remarkably different consequences for photosynthetic carbon metabolism in transgenic Arabidopsis thaliana. Plant J 23:759–770

    Article  PubMed  CAS  Google Scholar 

  • Tielens AG, Van der Meer P, van den Heuvel JM, van den Bergh SG (1991) The enigmatic presence of all gluconeogenic enzymes in Schistosoma mansoni adults. Parasitology 102(Pt 2):267–276

    Article  PubMed  CAS  Google Scholar 

  • Verhees CH, Akerboom J, Schiltz E, de Vos WM, van der Oost J (2002) Molecular and biochemical characterization of a distinct type of fructose-1,6-bisphosphatase from Pyrococcus furiosus. J Bacteriol 184:3401–3405

    Article  PubMed  CAS  Google Scholar 

  • Wang Y, Tomlinson B (2007) Managlinat dialanetil, a fructose-1,6-bisphosphatase inhibitor for the treatment of type 2 diabetes. Curr Opin Investig Drugs 8:849–858

    PubMed  CAS  Google Scholar 

  • Williamson HJ, Allardyce RA, Clemett RS, Hidajat RR (1990) Serum and neutrophils alter the rate of excretory/secretory antigen release by Toxocara canis infective larvae in vitro. Parasite Immunol 12:175–187

    Article  PubMed  CAS  Google Scholar 

  • Yamasaki H, Araki K, Lim PK, Zasmy N, Mak JW, Taib R, Aoki T (2000) Development of a highly specific recombinant Toxocara canis second-stage larva excretory-secretory antigen for immunodiagnosis of human toxocariasis. J Clin Microbiol 38:1409–1413

    PubMed  CAS  Google Scholar 

  • Yoon BI, Jung SY, Hur K, Lee JH, Joo KH, Lee YS, Kim DY (2000) Differentiation of hamster liver oval cell following Clonorchis sinensis infection. J Vet Med Sci 62:1303–1310

    Article  PubMed  CAS  Google Scholar 

  • Zhang X, Jin Z, Da R, Dong Y, Song W, Chen X, Huang Q, Ling H, Che Y, Li Y, Zhang F (2008) Fas/FasL-dependent apoptosis of hepatocytes induced in rat and patients with Clonorchis sinensis infection. Parasitol Res 103:393–399

    Article  PubMed  Google Scholar 

  • Zheng N, Xu J, Wu Z, Chen J, Hu X, Song L, Yang G, Ji C, Chen S, Gu S, Ying K, Yu X (2005) Clonorchis sinensis: molecular cloning and functional expression of novel cytosolic malate dehydrogenase. Exp Parasitol 109:220–227

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

This work was supported by National Science and Technology Major Project of China (No. 2008ZX10004-011), National Natural Science Foundation of China (No. 30671831), and National High Technology Research and Development Program of China (973 program, No. 2010CB530000).

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Authors and Affiliations

  1. Department of Parasitology, Key Laboratory for Tropical Diseases Control, Sun Yat-sen University, Guangzhou, 510080, People’s Republic of China

    Minghui Zheng, Xuchu Hu, Fengyu Hu, Lisi Huang, Peng Wang, Yue Hu, Yan Huang, Wenfang Li, Chi Liang & Xinbing Yu

  2. Key Laboratory for Tropical Diseases Control, Ministry of Education, Guangzhou, 510080, People’s Republic of China

    Minghui Zheng, Xuchu Hu, Fengyu Hu, Lisi Huang, Peng Wang, Yue Hu, Yan Huang, Wenfang Li, Chi Liang & Xinbing Yu

  3. Proteomics Center, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, 510080, People’s Republic of China

    Kunhua Hu

  4. Guangdong Provincial Key Laboratory of Liver Disease, The Third Affiliated Hospital, Sun Yat-sen University, Guangzhou, 510080, People’s Republic of China

    Wei Liu

  5. Institute of Life and Health Engineering, and National Engineering and Research Center for Genetic Medicine, Jinan University, Guangzhou, 510632, People’s Republic of China

    Xingfeng Yin & Qingyu He

  6. Institute of Infectious Diseases, Guangzhou No. 8 People’s Hospital, Guangzhou, 510080, People’s Republic of China

    Fengyu Hu

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  1. Minghui Zheng
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  2. Kunhua Hu
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  3. Wei Liu
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  7. Peng Wang
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  9. Yan Huang
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  10. Wenfang Li
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  11. Chi Liang
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Correspondence to Xinbing Yu.

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Zheng, M., Hu, K., Liu, W. et al. Proteomic analysis of excretory secretory products from Clonorchis sinensis adult worms: molecular characterization and serological reactivity of a excretory–secretory antigen-fructose-1,6-bisphosphatase. Parasitol Res 109, 737–744 (2011). https://doi.org/10.1007/s00436-011-2316-5

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