
Peptidomics of an in vitro digested α-Gal carrying protein revealed IgE-reactive peptides
- Author
- D Apostolovic, M Krstic, J Mihailovic, M Starkhammar, Tanja Cirkovic Velickovic (UGent) , C Hamsten and M van Hage
- Organization
- Abstract
- The mammalian carbohydrate galactose-alpha 1,3-galactose (alpha-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 alpha-Gal glycoproteins using an in vitro model. Bovine thyroglobulin (BTG), a typical alpha-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 alpha-Gal epitope on the obtained peptides was demonstrated by an anti-alpha-Gal antibody and IgE from red meat allergic patients. The alpha-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 alpha-Gal residues were identified at Asn(1756), Asn(1850) and Asn(2231). Thus allergenic a-Gal epitopes are stable to pepsinolysis, reinforcing their role as clinically relevant food allergens.
- Keywords
- MEAT-INDUCED ANAPHYLAXIS, RED MEAT, MAMMALIAN MEAT, GASTROINTESTINAL, DIGESTION, BOVINE THYROGLOBULIN, RESISTANT PEPTIDES, IMMUNOGLOBULIN-G, FOOD ALLERGENS, GALACTOSE-ALPHA-1, 3-GALACTOSE, STABILITY
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Citation
Please use this url to cite or link to this publication: http://hdl.handle.net/1854/LU-8615103
- MLA
- Apostolovic, D., et al. “Peptidomics of an in Vitro Digested α-Gal Carrying Protein Revealed IgE-Reactive Peptides.” SCIENTIFIC REPORTS, vol. 7, 2017, doi:10.1038/s41598-017-05355-4.
- APA
- 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. https://doi.org/10.1038/s41598-017-05355-4
- Chicago author-date
- Apostolovic, D, M Krstic, J Mihailovic, M Starkhammar, Tanja Cirkovic Velickovic, C Hamsten, and M van Hage. 2017. “Peptidomics of an in Vitro Digested α-Gal Carrying Protein Revealed IgE-Reactive Peptides.” SCIENTIFIC REPORTS 7. https://doi.org/10.1038/s41598-017-05355-4.
- Chicago author-date (all authors)
- Apostolovic, D, M Krstic, J Mihailovic, M Starkhammar, Tanja Cirkovic Velickovic, C Hamsten, and M van Hage. 2017. “Peptidomics of an in Vitro Digested α-Gal Carrying Protein Revealed IgE-Reactive Peptides.” SCIENTIFIC REPORTS 7. doi:10.1038/s41598-017-05355-4.
- Vancouver
- 1.Apostolovic D, Krstic M, Mihailovic J, Starkhammar M, Cirkovic Velickovic T, Hamsten C, et al. Peptidomics of an in vitro digested α-Gal carrying protein revealed IgE-reactive peptides. SCIENTIFIC REPORTS. 2017;7.
- IEEE
- [1]D. Apostolovic et al., “Peptidomics of an in vitro digested α-Gal carrying protein revealed IgE-reactive peptides,” SCIENTIFIC REPORTS, vol. 7, 2017.
@article{8615103, abstract = {{The mammalian carbohydrate galactose-alpha 1,3-galactose (alpha-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 alpha-Gal glycoproteins using an in vitro model. Bovine thyroglobulin (BTG), a typical alpha-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 alpha-Gal epitope on the obtained peptides was demonstrated by an anti-alpha-Gal antibody and IgE from red meat allergic patients. The alpha-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 alpha-Gal residues were identified at Asn(1756), Asn(1850) and Asn(2231). Thus allergenic a-Gal epitopes are stable to pepsinolysis, reinforcing their role as clinically relevant food allergens.}}, articleno = {{5201}}, author = {{Apostolovic, D and Krstic, M and Mihailovic, J and Starkhammar, M and Cirkovic Velickovic, Tanja and Hamsten, C and van Hage, M}}, issn = {{2045-2322}}, journal = {{SCIENTIFIC REPORTS}}, keywords = {{MEAT-INDUCED ANAPHYLAXIS,RED MEAT,MAMMALIAN MEAT,GASTROINTESTINAL,DIGESTION,BOVINE THYROGLOBULIN,RESISTANT PEPTIDES,IMMUNOGLOBULIN-G,FOOD ALLERGENS,GALACTOSE-ALPHA-1,3-GALACTOSE,STABILITY}}, language = {{eng}}, pages = {{10}}, title = {{Peptidomics of an in vitro digested α-Gal carrying protein revealed IgE-reactive peptides}}, url = {{http://doi.org/10.1038/s41598-017-05355-4}}, volume = {{7}}, year = {{2017}}, }
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