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Shrimp thrombospondin (TSP): presence of O-β1,4 N-acetylglucosamine polymers and its function in TSP chain association in egg extracellular matrix

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) w...

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Autores principales: Magerd, Sirilug, Senarai, Thanyaporn, Thongsum, Orawan, Chawiwithaya, Chidchanok, Sato, Chihiro, Kitajima, Ken, Weerachatyanukul, Wattana, Asuvapongpatana, Somluk, Surinlert, Piyaporn
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9106747/
https://www.ncbi.nlm.nih.gov/pubmed/35562392
http://dx.doi.org/10.1038/s41598-022-11873-7
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author Magerd, Sirilug
Senarai, Thanyaporn
Thongsum, Orawan
Chawiwithaya, Chidchanok
Sato, Chihiro
Kitajima, Ken
Weerachatyanukul, Wattana
Asuvapongpatana, Somluk
Surinlert, Piyaporn
author_facet Magerd, Sirilug
Senarai, Thanyaporn
Thongsum, Orawan
Chawiwithaya, Chidchanok
Sato, Chihiro
Kitajima, Ken
Weerachatyanukul, Wattana
Asuvapongpatana, Somluk
Surinlert, Piyaporn
author_sort Magerd, Sirilug
collection PubMed
description 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.
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spelling pubmed-91067472022-05-15 Shrimp thrombospondin (TSP): presence of O-β1,4 N-acetylglucosamine polymers and its function in TSP chain association in egg extracellular matrix Magerd, Sirilug Senarai, Thanyaporn Thongsum, Orawan Chawiwithaya, Chidchanok Sato, Chihiro Kitajima, Ken Weerachatyanukul, Wattana Asuvapongpatana, Somluk Surinlert, Piyaporn Sci Rep Article 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. Nature Publishing Group UK 2022-05-13 /pmc/articles/PMC9106747/ /pubmed/35562392 http://dx.doi.org/10.1038/s41598-022-11873-7 Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Article
Magerd, Sirilug
Senarai, Thanyaporn
Thongsum, Orawan
Chawiwithaya, Chidchanok
Sato, Chihiro
Kitajima, Ken
Weerachatyanukul, Wattana
Asuvapongpatana, Somluk
Surinlert, Piyaporn
Shrimp thrombospondin (TSP): presence of O-β1,4 N-acetylglucosamine polymers and its function in TSP chain association in egg extracellular matrix
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
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9106747/
https://www.ncbi.nlm.nih.gov/pubmed/35562392
http://dx.doi.org/10.1038/s41598-022-11873-7
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