Molecular characterization of a novel serine proteinase from Trichinella spiralis and its participation in larval invasion of gut epithelium.
<h4>Background</h4>A novel serine proteinase of Trichinells spiralis (TsSPc) has been identified in the excretion/secretion (ES) antigens, but its role in larval invasion is unclear. The aim of this study was to clone and express TsSPc, identify its biological and biochemical characteris...
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2023-09-01
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| Series: | PLoS Neglected Tropical Diseases |
| Online Access: | https://journals.plos.org/plosntds/article/file?id=10.1371/journal.pntd.0011629&type=printable |
| _version_ | 1827801007719972864 |
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| author | Yan Yan Song Xin Zhuo Zhang Bo Ning Wang Min Min Weng Zhao Yu Zhang Xin Guo Xi Zhang Zhong Quan Wang Jing Cui |
| author_facet | Yan Yan Song Xin Zhuo Zhang Bo Ning Wang Min Min Weng Zhao Yu Zhang Xin Guo Xi Zhang Zhong Quan Wang Jing Cui |
| author_sort | Yan Yan Song |
| collection | DOAJ |
| description | <h4>Background</h4>A novel serine proteinase of Trichinells spiralis (TsSPc) has been identified in the excretion/secretion (ES) antigens, but its role in larval invasion is unclear. The aim of this study was to clone and express TsSPc, identify its biological and biochemical characteristics, and investigate its role on larval invasion of gut epithelium during T. spiralis infection.<h4>Methodology/principal findings</h4>TsSPc has a functional domain of serine proteinase, and its tertiary structure consists of three amino acid residues (His88, Asp139 and Ser229) forming a pocket like functional domain. Recombinant TsSPc (rTsSPc) was expressed and purified. The rTsSPc has good immunogenicity. On Western blot analysis, rTsSPc was recognized by infection serum and anti-rTsSPc serum, natural TsSPc in crude and ES antigens was identified by anti-rTsSPc serum. The results of qPCR, Western blot and indirect immunofluorescence test (IIFT) showed that TsSPc was expressed at diverse stage worms, and mainly localized at cuticle, stichosome and intrauterine embryos of this nematode. The rTsSPc had enzymatic activity of native serine protease, which hydrolyzed the substrate BAEE, casein and collagen I. After site directed mutation of enzymatic active sites of TsSPc, its antigenicity did not change but the enzyme activity was fully lost. rTsSPc specifically bound to intestinal epithelium cells (IECs) and the binding sites were mainly localized in cell membrane and cytoplasm. rTsSPc accelerated larval invasion of IECs, whereas anti-rTsSPc antibodies and TsSPc-specific dsRNA obviously hindered larval invasion.<h4>Conclusions</h4>TsSPc was a surface and secretory proteinase of the parasite, participated in larval invasion of gut epithelium, and may be considered as a candidate vaccine target molecule against Trichinella intrusion and infection. |
| first_indexed | 2024-03-11T20:20:00Z |
| format | Article |
| id | doaj.art-037999ce83cc4f68981c26437c2e89a9 |
| institution | Directory Open Access Journal |
| issn | 1935-2727 1935-2735 |
| language | English |
| last_indexed | 2024-03-11T20:20:00Z |
| publishDate | 2023-09-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS Neglected Tropical Diseases |
| spelling | doaj.art-037999ce83cc4f68981c26437c2e89a92023-10-03T05:31:49ZengPublic Library of Science (PLoS)PLoS Neglected Tropical Diseases1935-27271935-27352023-09-01179e001162910.1371/journal.pntd.0011629Molecular characterization of a novel serine proteinase from Trichinella spiralis and its participation in larval invasion of gut epithelium.Yan Yan SongXin Zhuo ZhangBo Ning WangMin Min WengZhao Yu ZhangXin GuoXi ZhangZhong Quan WangJing Cui<h4>Background</h4>A novel serine proteinase of Trichinells spiralis (TsSPc) has been identified in the excretion/secretion (ES) antigens, but its role in larval invasion is unclear. The aim of this study was to clone and express TsSPc, identify its biological and biochemical characteristics, and investigate its role on larval invasion of gut epithelium during T. spiralis infection.<h4>Methodology/principal findings</h4>TsSPc has a functional domain of serine proteinase, and its tertiary structure consists of three amino acid residues (His88, Asp139 and Ser229) forming a pocket like functional domain. Recombinant TsSPc (rTsSPc) was expressed and purified. The rTsSPc has good immunogenicity. On Western blot analysis, rTsSPc was recognized by infection serum and anti-rTsSPc serum, natural TsSPc in crude and ES antigens was identified by anti-rTsSPc serum. The results of qPCR, Western blot and indirect immunofluorescence test (IIFT) showed that TsSPc was expressed at diverse stage worms, and mainly localized at cuticle, stichosome and intrauterine embryos of this nematode. The rTsSPc had enzymatic activity of native serine protease, which hydrolyzed the substrate BAEE, casein and collagen I. After site directed mutation of enzymatic active sites of TsSPc, its antigenicity did not change but the enzyme activity was fully lost. rTsSPc specifically bound to intestinal epithelium cells (IECs) and the binding sites were mainly localized in cell membrane and cytoplasm. rTsSPc accelerated larval invasion of IECs, whereas anti-rTsSPc antibodies and TsSPc-specific dsRNA obviously hindered larval invasion.<h4>Conclusions</h4>TsSPc was a surface and secretory proteinase of the parasite, participated in larval invasion of gut epithelium, and may be considered as a candidate vaccine target molecule against Trichinella intrusion and infection.https://journals.plos.org/plosntds/article/file?id=10.1371/journal.pntd.0011629&type=printable |
| spellingShingle | Yan Yan Song Xin Zhuo Zhang Bo Ning Wang Min Min Weng Zhao Yu Zhang Xin Guo Xi Zhang Zhong Quan Wang Jing Cui Molecular characterization of a novel serine proteinase from Trichinella spiralis and its participation in larval invasion of gut epithelium. PLoS Neglected Tropical Diseases |
| title | Molecular characterization of a novel serine proteinase from Trichinella spiralis and its participation in larval invasion of gut epithelium. |
| title_full | Molecular characterization of a novel serine proteinase from Trichinella spiralis and its participation in larval invasion of gut epithelium. |
| title_fullStr | Molecular characterization of a novel serine proteinase from Trichinella spiralis and its participation in larval invasion of gut epithelium. |
| title_full_unstemmed | Molecular characterization of a novel serine proteinase from Trichinella spiralis and its participation in larval invasion of gut epithelium. |
| title_short | Molecular characterization of a novel serine proteinase from Trichinella spiralis and its participation in larval invasion of gut epithelium. |
| title_sort | molecular characterization of a novel serine proteinase from trichinella spiralis and its participation in larval invasion of gut epithelium |
| url | https://journals.plos.org/plosntds/article/file?id=10.1371/journal.pntd.0011629&type=printable |
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