Cloning and characterization of a novel sigma-like glutathione S-transferase from the giant panda parasitic nematode, Baylisascaris schroederi
Abstract Background Baylisascaris schroederi, an intestinal nematode of the giant panda, is the cause of the often fatal disease, baylisascariasis. Glutathione S-transferases (GSTs) are versatile enzymes that can affect parasite survival and parasite-host interactions and, are therefore, potential t...
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| Format: | Article |
| Language: | English |
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BMC
2015-01-01
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| Series: | Parasites & Vectors |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s13071-014-0629-9 |
| _version_ | 1797811819046240256 |
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| author | Yue Xie Xuan Zhou Lin Chen Zhihe Zhang Chengdong Wang Xiaobin Gu Tao Wang Xuerong Peng Guangyou Yang |
| author_facet | Yue Xie Xuan Zhou Lin Chen Zhihe Zhang Chengdong Wang Xiaobin Gu Tao Wang Xuerong Peng Guangyou Yang |
| author_sort | Yue Xie |
| collection | DOAJ |
| description | Abstract Background Baylisascaris schroederi, an intestinal nematode of the giant panda, is the cause of the often fatal disease, baylisascariasis. Glutathione S-transferases (GSTs) are versatile enzymes that can affect parasite survival and parasite-host interactions and, are therefore, potential targets for the development of diagnostic tests and vaccines. Methods In this study, we identified a full-length cDNA that encoded a novel, secretory sigma-like GST (Bsc-GSTσ) from a B. schroederi-omic dataset. Following cloning and sequencing, sequence and structural analyses and comparative modeling were performed using online-bioinformatics and proteomics tools. The recombinant Bsc-GSTσ (rBsc-GSTσ) protein was prokaryotically expressed and then used to detect antigenicity and reactivity using immunoblotting assays. In addition, the native protein in female adult B. schroederi was located via immunofluorescence techniques, while the preliminary ELISA-based serodiagnostic potential of rBsc-GSTσ was assessed in native and infected mouse sera. Results Bsc-GSTσ contained a 621-bp open reading frame that encoded a polypeptide of 206 amino acids with two typical sigma GST domain profiles, including a GST_N_Sigma_like at the N-terminus and a GST_C_Sigma_like at the C-terminus. The presence of an N-terminal signal sequence indicated that Bsc-GSTσ was a secretory protein. Sequence alignment and phylogenetic analyses showed that Bsc-GSTσ was a nematode-specific member of the Sigma class GSTs and shared the closest genetic distance with its homologue in Ascaris suum. Further comparative structure analyses indicated that Bsc-GSTσ possessed the essential structural motifs (e.g., βαβαββα) and the consensus secondary or tertiary structure that is typical for other characterized GSTσs. Immunolocalization revealed strong distributions of native Bsc-GSTσ in the body hypodermis, lateral chords, gut epithelium, gut microvilli, oviduct epithelium, and ovaries of adult female worms, similar to its homologue in A. suum. Building on good immunogenic properties, rBsc-GSTσ-based ELISA exhibited a sensitivity of 79.1% and a specificity of 82.0% to detect anti-B. schroederi IgG antibodies in the sera of experimentally infected mice. Conclusion This study presents a comprehensive demonstration of sequence and structural-based analysis of a new, secretory sigma-like GST from a nematode, and its good serodiagnostic performance suggests that rBsc-GSTσ has the potential to detect B. schroederi and, therefore, could be used to develop an ELISA-based serological test to diagnose baylisascariasis in giant pandas. |
| first_indexed | 2024-03-13T07:28:23Z |
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| institution | Directory Open Access Journal |
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| language | English |
| last_indexed | 2024-03-13T07:28:23Z |
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| series | Parasites & Vectors |
| spelling | doaj.art-6c1c11a7788042709259b0edac91d5032023-06-04T11:13:16ZengBMCParasites & Vectors1756-33052015-01-018111310.1186/s13071-014-0629-9Cloning and characterization of a novel sigma-like glutathione S-transferase from the giant panda parasitic nematode, Baylisascaris schroederiYue Xie0Xuan Zhou1Lin Chen2Zhihe Zhang3Chengdong Wang4Xiaobin Gu5Tao Wang6Xuerong Peng7Guangyou Yang8Department of Parasitology, College of Veterinary Medicine, Sichuan Agricultural UniversityCentre for Animal Diseases Control and Prevention, Dachuan Animal Husbandry BureauDepartment of Parasitology, College of Veterinary Medicine, Sichuan Agricultural UniversityChengdu Research Base of Giant Panda BreedingChina Conservation and Research Center for Giant PandaDepartment of Parasitology, College of Veterinary Medicine, Sichuan Agricultural UniversityDepartment of Parasitology, College of Veterinary Medicine, Sichuan Agricultural UniversityDepartment of Chemistry, College of Life and Basic Science, Sichuan Agricultural UniversityDepartment of Parasitology, College of Veterinary Medicine, Sichuan Agricultural UniversityAbstract Background Baylisascaris schroederi, an intestinal nematode of the giant panda, is the cause of the often fatal disease, baylisascariasis. Glutathione S-transferases (GSTs) are versatile enzymes that can affect parasite survival and parasite-host interactions and, are therefore, potential targets for the development of diagnostic tests and vaccines. Methods In this study, we identified a full-length cDNA that encoded a novel, secretory sigma-like GST (Bsc-GSTσ) from a B. schroederi-omic dataset. Following cloning and sequencing, sequence and structural analyses and comparative modeling were performed using online-bioinformatics and proteomics tools. The recombinant Bsc-GSTσ (rBsc-GSTσ) protein was prokaryotically expressed and then used to detect antigenicity and reactivity using immunoblotting assays. In addition, the native protein in female adult B. schroederi was located via immunofluorescence techniques, while the preliminary ELISA-based serodiagnostic potential of rBsc-GSTσ was assessed in native and infected mouse sera. Results Bsc-GSTσ contained a 621-bp open reading frame that encoded a polypeptide of 206 amino acids with two typical sigma GST domain profiles, including a GST_N_Sigma_like at the N-terminus and a GST_C_Sigma_like at the C-terminus. The presence of an N-terminal signal sequence indicated that Bsc-GSTσ was a secretory protein. Sequence alignment and phylogenetic analyses showed that Bsc-GSTσ was a nematode-specific member of the Sigma class GSTs and shared the closest genetic distance with its homologue in Ascaris suum. Further comparative structure analyses indicated that Bsc-GSTσ possessed the essential structural motifs (e.g., βαβαββα) and the consensus secondary or tertiary structure that is typical for other characterized GSTσs. Immunolocalization revealed strong distributions of native Bsc-GSTσ in the body hypodermis, lateral chords, gut epithelium, gut microvilli, oviduct epithelium, and ovaries of adult female worms, similar to its homologue in A. suum. Building on good immunogenic properties, rBsc-GSTσ-based ELISA exhibited a sensitivity of 79.1% and a specificity of 82.0% to detect anti-B. schroederi IgG antibodies in the sera of experimentally infected mice. Conclusion This study presents a comprehensive demonstration of sequence and structural-based analysis of a new, secretory sigma-like GST from a nematode, and its good serodiagnostic performance suggests that rBsc-GSTσ has the potential to detect B. schroederi and, therefore, could be used to develop an ELISA-based serological test to diagnose baylisascariasis in giant pandas.https://doi.org/10.1186/s13071-014-0629-9Baylisascaris schroederiGiant pandaGlutathione S-transferaseSigma classBsc-GSTσImmunolocalization |
| spellingShingle | Yue Xie Xuan Zhou Lin Chen Zhihe Zhang Chengdong Wang Xiaobin Gu Tao Wang Xuerong Peng Guangyou Yang Cloning and characterization of a novel sigma-like glutathione S-transferase from the giant panda parasitic nematode, Baylisascaris schroederi Parasites & Vectors Baylisascaris schroederi Giant panda Glutathione S-transferase Sigma class Bsc-GSTσ Immunolocalization |
| title | Cloning and characterization of a novel sigma-like glutathione S-transferase from the giant panda parasitic nematode, Baylisascaris schroederi |
| title_full | Cloning and characterization of a novel sigma-like glutathione S-transferase from the giant panda parasitic nematode, Baylisascaris schroederi |
| title_fullStr | Cloning and characterization of a novel sigma-like glutathione S-transferase from the giant panda parasitic nematode, Baylisascaris schroederi |
| title_full_unstemmed | Cloning and characterization of a novel sigma-like glutathione S-transferase from the giant panda parasitic nematode, Baylisascaris schroederi |
| title_short | Cloning and characterization of a novel sigma-like glutathione S-transferase from the giant panda parasitic nematode, Baylisascaris schroederi |
| title_sort | cloning and characterization of a novel sigma like glutathione s transferase from the giant panda parasitic nematode baylisascaris schroederi |
| topic | Baylisascaris schroederi Giant panda Glutathione S-transferase Sigma class Bsc-GSTσ Immunolocalization |
| url | https://doi.org/10.1186/s13071-014-0629-9 |
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