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

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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:

MLA
Apostolovic, D et al. “Peptidomics of an in Vitro Digested α-Gal Carrying Protein Revealed IgE-reactive Peptides.” SCIENTIFIC REPORTS 7 (2017): n. pag. Print.
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.
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.
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.
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://dx.doi.org/10.1038/s41598-017-05355-4},
  volume       = {7},
  year         = {2017},
}

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