Identification of a serodiagnostic antigen,legumain, by immunoproteomic analysis of excretory‐secretory products of Clonorchis sinensis adult worms |
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Authors: | Jung‐Won Ju Hyun‐Na Joo Myoung‐Ro Lee Shin‐Hyeong Cho Hyeng‐Il Cheun Jung‐Yeon Kim Young‐Hee Lee Kwang‐Jun Lee Woon‐Mok Sohn Dong‐Min Kim Il‐Chul Kim Byoung Chul Park Tong‐Soo Kim |
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Affiliation: | 1. Division of Malaria and Parasitic Diseases, National Institute of Health, Seoul, Korea;2. Lab of Pathogenic Proteomics, National Institute of Health, Seoul, Korea;3. Department of Parasitology and Institute of Health Sciences, College of Medicine, Gyeongsang National University, Jinju, Korea;4. BK21 Ubiquitous Information Appliances, Chonnam National University, Gwangju, Korea;5. Department of Biology (the Second Stage BK21 Program), College of Natural Sciences, Chonnam National University, Gwangju, Korea;6. Division of Translational Research, Korea Research Institute of Bioscience and Biotechnology, Daejeon, Korea;7. Department of Parasitology, Inha Research Institute for Medical Sciences, Inha University College of Medicine, Incheon, Korea |
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Abstract: | Clonorchis sinensis, the Chinese liver fluke, is the causative agent of clonorchiasis as well as liver and biliary diseases. The excretory‐secretory products (ESPs) of the parasites play important roles in host–parasite interactions. In this study, we have investigated the proteome of ESPs obtained from C. sinensis adult worms. Although the full genome database of C. sinensis is not yet available, we have successfully identified 62 protein spots using 2‐DE‐based mass analysis and EST database of C. sinensis. The proteins identified include detoxification enzymes, such as glutathione S‐transferase and thioredoxin peroxidase, myoglobin and a number of cysteine proteases that are expressed abundantly. In order to identify potential targets for the diagnosis and therapy of clonorchiasis, we conducted immunoblot analysis of the ESPs proteome using the sera obtained from clonorchiasis patients and identified legumains and cysteine proteases as antigens present in the ESPs. Although the cysteine proteases were previously reported to elicit antigenicity, the legumains are found herein for the first time as a serological antigen of C. sinensis. To confirm these findings, we expressed recombinant legumain in Escherichia coli and verified that recombinant legumain also functions as a potent antigen against the sera of clonorchiasis patients. Our results illustrate the validity of immuno‐proteomic approaches in the identification of serodiagnostic antigens in the parasites. |
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Keywords: | Antigen Clonorchis sinensis Excretory‐secretory products Legumain Proteome |
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