Tags

Type your tag names separated by a space and hit enter

Peptidomics of an in vitro digested α-Gal carrying protein revealed IgE-reactive peptides.
Sci Rep. 2017 07 12; 7(1):5201.SR

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.

Authors+Show Affiliations

Department of Medicine Solna, Immunology and Allergy Unit, Karolinska Institutet and Karolinska University Hospital, Stockholm, Sweden.Department of Medicine Solna, Immunology and Allergy Unit, Karolinska Institutet and Karolinska University Hospital, Stockholm, Sweden. Center of Excellence for Molecular Food Sciences, University of Belgrade, Faculty of Chemistry, Belgrade, Serbia.Center of Excellence for Molecular Food Sciences, University of Belgrade, Faculty of Chemistry, Belgrade, Serbia.Department of Internal Medicine, Södersjukhuset, Stockholm, Sweden.Center of Excellence for Molecular Food Sciences, University of Belgrade, Faculty of Chemistry, Belgrade, Serbia. Ghent University Global Campus, Yeonsu-gu, Incheon, South Korea. Faculty of Bioscience Engineering, Ghent University, Ghent, Belgium.Department of Medicine Solna, Immunology and Allergy Unit, Karolinska Institutet and Karolinska University Hospital, Stockholm, Sweden.Department of Medicine Solna, Immunology and Allergy Unit, Karolinska Institutet and Karolinska University Hospital, Stockholm, Sweden. marianne.van.hage@ki.se.

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

28701697

Citation

Apostolovic, D, et al. "Peptidomics of an in Vitro Digested α-Gal Carrying Protein Revealed IgE-reactive Peptides." Scientific Reports, vol. 7, no. 1, 2017, p. 5201.
Apostolovic D, Krstic M, Mihailovic J, et al. Peptidomics of an in vitro digested α-Gal carrying protein revealed IgE-reactive peptides. Sci Rep. 2017;7(1):5201.
Apostolovic, D., Krstic, M., Mihailovic, J., Starkhammar, M., Cirkovic Velickovic, T., Hamsten, C., & van Hage, M. (2017). Peptidomics of an in vitro digested α-Gal carrying protein revealed IgE-reactive peptides. Scientific Reports, 7(1), 5201. https://doi.org/10.1038/s41598-017-05355-4
Apostolovic D, et al. Peptidomics of an in Vitro Digested α-Gal Carrying Protein Revealed IgE-reactive Peptides. Sci Rep. 2017 07 12;7(1):5201. PubMed PMID: 28701697.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Peptidomics of an in vitro digested α-Gal carrying protein revealed IgE-reactive peptides. AU - Apostolovic,D, AU - Krstic,M, AU - Mihailovic,J, AU - Starkhammar,M, AU - Cirkovic Velickovic,T, AU - Hamsten,C, AU - van Hage,M, Y1 - 2017/07/12/ PY - 2017/02/23/received PY - 2017/05/26/accepted PY - 2017/7/14/entrez PY - 2017/7/14/pubmed PY - 2019/2/7/medline SP - 5201 EP - 5201 JF - Scientific reports JO - Sci Rep VL - 7 IS - 1 N2 - 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. SN - 2045-2322 UR - https://www.unboundmedicine.com/medline/citation/28701697/Peptidomics_of_an_in_vitro_digested_α_Gal_carrying_protein_revealed_IgE_reactive_peptides_ L2 - http://dx.doi.org/10.1038/s41598-017-05355-4 DB - PRIME DP - Unbound Medicine ER -