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


ABSTRACT: 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-?-1,3-galactose (?-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 ?-Gal carrying proteins in the gastrointestinal tract. The aim of this study was to investigate how protein glycosylation by ?-Gal affects the susceptibility to gastric digestion and transport through the Caco-2 cell monolayer. We found that ?-Gal glycosylation altered protein susceptibility to gastric digestion, where large protein fragments bearing the ?-Gal epitope remained for up to 2 h of digestion. Furthermore, ?-Gal glycosylation of the protein hampered transcytosis of the protein through the Caco-2 monolayer. ?-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 ?-Gal glycosylated BSA (bovine serum albumin) (BSA-?-Gal). Taken together, our data add new knowledge and shed light on the digestion and transport of ?-Gal glycosylated proteins.

SUBMITTER: Krstic Ristivojevic M 

PROVIDER: S-EPMC7461108 | BioStudies | 2020-01-01

REPOSITORIES: biostudies

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