Shrimp thrombospondin (TSP): presence of O-β1,4 N-acetylglucosamine polymers and its function in TSP chain association in egg extracellular matrix
Abstract We characterized the existence of O-β(1,4)-GlcNAc polymers (β1,4GNP) that were anchored on the O-linked glycosylation sites of shrimp thrombospondin (pmTSP-II). There were five putative β1,4GNP linkages on the epithelial growth factor-like domain of pmTSP-II. Antibody against O-β-GlcNAc (CT...
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2022-05-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-022-11873-7 |
| _version_ | 1811233709240614912 |
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| author | Sirilug Magerd Thanyaporn Senarai Orawan Thongsum Chidchanok Chawiwithaya Chihiro Sato Ken Kitajima Wattana Weerachatyanukul Somluk Asuvapongpatana Piyaporn Surinlert |
| author_facet | Sirilug Magerd Thanyaporn Senarai Orawan Thongsum Chidchanok Chawiwithaya Chihiro Sato Ken Kitajima Wattana Weerachatyanukul Somluk Asuvapongpatana Piyaporn Surinlert |
| author_sort | Sirilug Magerd |
| collection | DOAJ |
| description | Abstract We characterized the existence of O-β(1,4)-GlcNAc polymers (β1,4GNP) that were anchored on the O-linked glycosylation sites of shrimp thrombospondin (pmTSP-II). There were five putative β1,4GNP linkages on the epithelial growth factor-like domain of pmTSP-II. Antibody against O-β-GlcNAc (CTD110.6) was used to prove the existence of linear and complex β1,4GNP. The antibody well reacted with linear chito-triose, -tetraose and -pentaose conjugated with phosphatidylethanolamine lipid. The immunoreactivity could also be detected with a complex β1,4GNP within pmTSP-II (at MW > 250 kDa). Upon denaturing the protein with SDS-PAGE buffer, the size of pmTSP-II was shifted to be 250 kDa, approximately 2.5 folds larger than the deduced molecular mass of pmTSP-II (110 kDa), suggesting additional association of pmTSP-II apart from its known disulfide bridging. This was confirmed by chitinase digestion on pmTSP-II protein leading to the subsequent smaller protein bands at 110–170 kDa in time- and concentration-dependent manners. These bands well reacted with CTD110.6 antibody and disappeared after extensive chitinase hydrolysis. Together, we believe that β1,4GNP on pmTSP-II serve the function in an inter-chain association to provide structural architecture of egg extracellular matrix, a novel function of pmTSP-II in reproductive biology. |
| first_indexed | 2024-04-12T11:24:07Z |
| format | Article |
| id | doaj.art-4dfdb5e41b6747dcbf0dae1875da31a1 |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-04-12T11:24:07Z |
| publishDate | 2022-05-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-4dfdb5e41b6747dcbf0dae1875da31a12022-12-22T03:35:16ZengNature PortfolioScientific Reports2045-23222022-05-0112111010.1038/s41598-022-11873-7Shrimp thrombospondin (TSP): presence of O-β1,4 N-acetylglucosamine polymers and its function in TSP chain association in egg extracellular matrixSirilug Magerd0Thanyaporn Senarai1Orawan Thongsum2Chidchanok Chawiwithaya3Chihiro Sato4Ken Kitajima5Wattana Weerachatyanukul6Somluk Asuvapongpatana7Piyaporn Surinlert8Department of Basic Medical Science, Faculty of Medicine Vajira Hospital, Navamindradhiraj UniversityDepartment of Anatomy, Faculty of Medicine, Khon Kaen UniversityDepartment of Anatomy, Faculty of Science, Mahidol UniversityDepartment of Anatomy, Faculty of Science, Mahidol UniversityBioscience and Biotechnology Center and Graduate School of Bioagricultural Sciences, Nagoya UniversityBioscience and Biotechnology Center and Graduate School of Bioagricultural Sciences, Nagoya UniversityDepartment of Anatomy, Faculty of Science, Mahidol UniversityDepartment of Anatomy, Faculty of Science, Mahidol UniversityChulabhorn International College of Medicine, Thammasat UniversityAbstract We characterized the existence of O-β(1,4)-GlcNAc polymers (β1,4GNP) that were anchored on the O-linked glycosylation sites of shrimp thrombospondin (pmTSP-II). There were five putative β1,4GNP linkages on the epithelial growth factor-like domain of pmTSP-II. Antibody against O-β-GlcNAc (CTD110.6) was used to prove the existence of linear and complex β1,4GNP. The antibody well reacted with linear chito-triose, -tetraose and -pentaose conjugated with phosphatidylethanolamine lipid. The immunoreactivity could also be detected with a complex β1,4GNP within pmTSP-II (at MW > 250 kDa). Upon denaturing the protein with SDS-PAGE buffer, the size of pmTSP-II was shifted to be 250 kDa, approximately 2.5 folds larger than the deduced molecular mass of pmTSP-II (110 kDa), suggesting additional association of pmTSP-II apart from its known disulfide bridging. This was confirmed by chitinase digestion on pmTSP-II protein leading to the subsequent smaller protein bands at 110–170 kDa in time- and concentration-dependent manners. These bands well reacted with CTD110.6 antibody and disappeared after extensive chitinase hydrolysis. Together, we believe that β1,4GNP on pmTSP-II serve the function in an inter-chain association to provide structural architecture of egg extracellular matrix, a novel function of pmTSP-II in reproductive biology.https://doi.org/10.1038/s41598-022-11873-7 |
| spellingShingle | Sirilug Magerd Thanyaporn Senarai Orawan Thongsum Chidchanok Chawiwithaya Chihiro Sato Ken Kitajima Wattana Weerachatyanukul Somluk Asuvapongpatana Piyaporn Surinlert Shrimp thrombospondin (TSP): presence of O-β1,4 N-acetylglucosamine polymers and its function in TSP chain association in egg extracellular matrix Scientific Reports |
| title | Shrimp thrombospondin (TSP): presence of O-β1,4 N-acetylglucosamine polymers and its function in TSP chain association in egg extracellular matrix |
| title_full | Shrimp thrombospondin (TSP): presence of O-β1,4 N-acetylglucosamine polymers and its function in TSP chain association in egg extracellular matrix |
| title_fullStr | Shrimp thrombospondin (TSP): presence of O-β1,4 N-acetylglucosamine polymers and its function in TSP chain association in egg extracellular matrix |
| title_full_unstemmed | Shrimp thrombospondin (TSP): presence of O-β1,4 N-acetylglucosamine polymers and its function in TSP chain association in egg extracellular matrix |
| title_short | Shrimp thrombospondin (TSP): presence of O-β1,4 N-acetylglucosamine polymers and its function in TSP chain association in egg extracellular matrix |
| title_sort | shrimp thrombospondin tsp presence of o β1 4 n acetylglucosamine polymers and its function in tsp chain association in egg extracellular matrix |
| url | https://doi.org/10.1038/s41598-022-11873-7 |
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