Peptidomics of an in vitro digested α-Gal carrying protein revealed IgE-reactive peptides

Abstract The mammalian carbohydrate galactose-α1,3-galactose (α-Gal) causes a novel form of food allergy, red meat allergy, where patients experience severe allergic reactions several hours after red meat consumption. Here we explored gastric digestion of α-Gal glycoproteins using an in vitro model....

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Autores principales:	D. Apostolovic, M. Krstic, J. Mihailovic, M. Starkhammar, T. Cirkovic Velickovic, C. Hamsten, M. van Hage
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/5df1d610fcf444c1a7cf11ab75f2bb68
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spelling	oai:doaj.org-article:5df1d610fcf444c1a7cf11ab75f2bb682021-12-02T12:30:24ZPeptidomics of an in vitro digested α-Gal carrying protein revealed IgE-reactive peptides10.1038/s41598-017-05355-42045-2322https://doaj.org/article/5df1d610fcf444c1a7cf11ab75f2bb682017-07-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-05355-4https://doaj.org/toc/2045-2322Abstract The mammalian carbohydrate galactose-α1,3-galactose (α-Gal) causes a novel form of food allergy, red meat allergy, where patients experience severe allergic reactions several hours after red meat consumption. Here we explored gastric digestion of α-Gal glycoproteins using an in vitro model. Bovine thyroglobulin (BTG), a typical α-Gal carrying glycoprotein, was digested with pepsin. The resulting peptides were characterised by SDS PAGE, immunoblot and ImmunoCAP using sera from 20 red meat allergic patients. During pepsinolysis of BTG, a wide range of peptide bands was observed of which 14 to 17 kDa peptides remained stable throughout the gastric phase. The presence of the α-Gal epitope on the obtained peptides was demonstrated by an anti-α-Gal antibody and IgE from red meat allergic patients. The α-Gal digests were able to inhibit up to 86% of IgE reactivity to BTG. Importantly, basophil activation test demonstrated that the allergenic activity of BTG was retained after digestion in all four tested patients. Mass spectrometry-based peptidomics revealed that these peptides represent mostly internal and C-terminal parts of the protein, where the most potent IgE-binding α-Gal residues were identified at Asn1756, Asn1850 and Asn2231. Thus allergenic α-Gal epitopes are stable to pepsinolysis, reinforcing their role as clinically relevant food allergens.D. ApostolovicM. KrsticJ. MihailovicM. StarkhammarT. Cirkovic VelickovicC. HamstenM. van HageNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-10 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q D. Apostolovic M. Krstic J. Mihailovic M. Starkhammar T. Cirkovic Velickovic C. Hamsten M. van Hage Peptidomics of an in vitro digested α-Gal carrying protein revealed IgE-reactive peptides
description	Abstract The mammalian carbohydrate galactose-α1,3-galactose (α-Gal) causes a novel form of food allergy, red meat allergy, where patients experience severe allergic reactions several hours after red meat consumption. Here we explored gastric digestion of α-Gal glycoproteins using an in vitro model. Bovine thyroglobulin (BTG), a typical α-Gal carrying glycoprotein, was digested with pepsin. The resulting peptides were characterised by SDS PAGE, immunoblot and ImmunoCAP using sera from 20 red meat allergic patients. During pepsinolysis of BTG, a wide range of peptide bands was observed of which 14 to 17 kDa peptides remained stable throughout the gastric phase. The presence of the α-Gal epitope on the obtained peptides was demonstrated by an anti-α-Gal antibody and IgE from red meat allergic patients. The α-Gal digests were able to inhibit up to 86% of IgE reactivity to BTG. Importantly, basophil activation test demonstrated that the allergenic activity of BTG was retained after digestion in all four tested patients. Mass spectrometry-based peptidomics revealed that these peptides represent mostly internal and C-terminal parts of the protein, where the most potent IgE-binding α-Gal residues were identified at Asn1756, Asn1850 and Asn2231. Thus allergenic α-Gal epitopes are stable to pepsinolysis, reinforcing their role as clinically relevant food allergens.
format	article
author	D. Apostolovic M. Krstic J. Mihailovic M. Starkhammar T. Cirkovic Velickovic C. Hamsten M. van Hage
author_facet	D. Apostolovic M. Krstic J. Mihailovic M. Starkhammar T. Cirkovic Velickovic C. Hamsten M. van Hage
author_sort	D. Apostolovic
title	Peptidomics of an in vitro digested α-Gal carrying protein revealed IgE-reactive peptides
title_short	Peptidomics of an in vitro digested α-Gal carrying protein revealed IgE-reactive peptides
title_full	Peptidomics of an in vitro digested α-Gal carrying protein revealed IgE-reactive peptides
title_fullStr	Peptidomics of an in vitro digested α-Gal carrying protein revealed IgE-reactive peptides
title_full_unstemmed	Peptidomics of an in vitro digested α-Gal carrying protein revealed IgE-reactive peptides
title_sort	peptidomics of an in vitro digested α-gal carrying protein revealed ige-reactive peptides
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/5df1d610fcf444c1a7cf11ab75f2bb68
work_keys_str_mv	AT dapostolovic peptidomicsofaninvitrodigestedagalcarryingproteinrevealedigereactivepeptides AT mkrstic peptidomicsofaninvitrodigestedagalcarryingproteinrevealedigereactivepeptides AT jmihailovic peptidomicsofaninvitrodigestedagalcarryingproteinrevealedigereactivepeptides AT mstarkhammar peptidomicsofaninvitrodigestedagalcarryingproteinrevealedigereactivepeptides AT tcirkovicvelickovic peptidomicsofaninvitrodigestedagalcarryingproteinrevealedigereactivepeptides AT chamsten peptidomicsofaninvitrodigestedagalcarryingproteinrevealedigereactivepeptides AT mvanhage peptidomicsofaninvitrodigestedagalcarryingproteinrevealedigereactivepeptides
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Peptidomics of an in vitro digested α-Gal carrying protein revealed IgE-reactive peptides

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