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Alpha-Gal on the Protein Surface Hampers Transcytosis through the Caco-2 Monolayer

Transepithelial transport of proteins is an important step in the immune response to food allergens. Mammalian meat allergy is characterized by an IgE response against the carbohydrate moiety galactosyl-α-1,3-galactose (α-Gal) present on mammalian glycoproteins and glycolipids, which causes severe a...

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Autores principales: Krstić Ristivojević, Maja, Grundström, Jeanette, Apostolović, Danijela, Radomirović, Mirjana, Jovanović, Vesna, Radoi, Vlad, Kiewiet, M. B. Gea, Vukojević, Vladana, Ćirković Veličković, Tanja, van Hage, Marianne
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7461108/
https://www.ncbi.nlm.nih.gov/pubmed/32796496
http://dx.doi.org/10.3390/ijms21165742
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author Krstić Ristivojević, Maja
Grundström, Jeanette
Apostolović, Danijela
Radomirović, Mirjana
Jovanović, Vesna
Radoi, Vlad
Kiewiet, M. B. Gea
Vukojević, Vladana
Ćirković Veličković, Tanja
van Hage, Marianne
author_facet Krstić Ristivojević, Maja
Grundström, Jeanette
Apostolović, Danijela
Radomirović, Mirjana
Jovanović, Vesna
Radoi, Vlad
Kiewiet, M. B. Gea
Vukojević, Vladana
Ćirković Veličković, Tanja
van Hage, Marianne
author_sort Krstić Ristivojević, Maja
collection PubMed
description Transepithelial transport of proteins is an important step in the immune response to food allergens. Mammalian meat allergy is characterized by an IgE response against the carbohydrate moiety galactosyl-α-1,3-galactose (α-Gal) present on mammalian glycoproteins and glycolipids, which causes severe allergic reactions several hours after red meat consumption. The delayed reaction may be related to the processing of α-Gal carrying proteins in the gastrointestinal tract. The aim of this study was to investigate how protein glycosylation by α-Gal affects the susceptibility to gastric digestion and transport through the Caco-2 cell monolayer. We found that α-Gal glycosylation altered protein susceptibility to gastric digestion, where large protein fragments bearing the α-Gal epitope remained for up to 2 h of digestion. Furthermore, α-Gal glycosylation of the protein hampered transcytosis of the protein through the Caco-2 monolayer. α-Gal epitope on the intact protein could be detected in the endosomal fraction obtained by differential centrifugation of Caco-2 cell lysates. Furthermore, the level of galectin-3 in Caco-2 cells was not affected by the presence of α-Gal glycosylated BSA (bovine serum albumin) (BSA-α-Gal). Taken together, our data add new knowledge and shed light on the digestion and transport of α-Gal glycosylated proteins.
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spelling pubmed-74611082020-09-14 Alpha-Gal on the Protein Surface Hampers Transcytosis through the Caco-2 Monolayer Krstić Ristivojević, Maja Grundström, Jeanette Apostolović, Danijela Radomirović, Mirjana Jovanović, Vesna Radoi, Vlad Kiewiet, M. B. Gea Vukojević, Vladana Ćirković Veličković, Tanja van Hage, Marianne Int J Mol Sci Article Transepithelial transport of proteins is an important step in the immune response to food allergens. Mammalian meat allergy is characterized by an IgE response against the carbohydrate moiety galactosyl-α-1,3-galactose (α-Gal) present on mammalian glycoproteins and glycolipids, which causes severe allergic reactions several hours after red meat consumption. The delayed reaction may be related to the processing of α-Gal carrying proteins in the gastrointestinal tract. The aim of this study was to investigate how protein glycosylation by α-Gal affects the susceptibility to gastric digestion and transport through the Caco-2 cell monolayer. We found that α-Gal glycosylation altered protein susceptibility to gastric digestion, where large protein fragments bearing the α-Gal epitope remained for up to 2 h of digestion. Furthermore, α-Gal glycosylation of the protein hampered transcytosis of the protein through the Caco-2 monolayer. α-Gal epitope on the intact protein could be detected in the endosomal fraction obtained by differential centrifugation of Caco-2 cell lysates. Furthermore, the level of galectin-3 in Caco-2 cells was not affected by the presence of α-Gal glycosylated BSA (bovine serum albumin) (BSA-α-Gal). Taken together, our data add new knowledge and shed light on the digestion and transport of α-Gal glycosylated proteins. MDPI 2020-08-11 /pmc/articles/PMC7461108/ /pubmed/32796496 http://dx.doi.org/10.3390/ijms21165742 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Krstić Ristivojević, Maja
Grundström, Jeanette
Apostolović, Danijela
Radomirović, Mirjana
Jovanović, Vesna
Radoi, Vlad
Kiewiet, M. B. Gea
Vukojević, Vladana
Ćirković Veličković, Tanja
van Hage, Marianne
Alpha-Gal on the Protein Surface Hampers Transcytosis through the Caco-2 Monolayer
title Alpha-Gal on the Protein Surface Hampers Transcytosis through the Caco-2 Monolayer
title_full Alpha-Gal on the Protein Surface Hampers Transcytosis through the Caco-2 Monolayer
title_fullStr Alpha-Gal on the Protein Surface Hampers Transcytosis through the Caco-2 Monolayer
title_full_unstemmed Alpha-Gal on the Protein Surface Hampers Transcytosis through the Caco-2 Monolayer
title_short Alpha-Gal on the Protein Surface Hampers Transcytosis through the Caco-2 Monolayer
title_sort alpha-gal on the protein surface hampers transcytosis through the caco-2 monolayer
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7461108/
https://www.ncbi.nlm.nih.gov/pubmed/32796496
http://dx.doi.org/10.3390/ijms21165742
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