Peptidomics of an in vitro digested alpha-Gal carrying protein revealed IgE-reactive peptides
2017
Аутори
Apostolović, DanijelaKrstić-Ristivojević, Maja
Mihailović-Vesić, Jelena
Starkhammar, Maria
Ćirković-Veličković, Tanja
Hamsten, Carl
van Hage, Marianne
Чланак у часопису (Објављена верзија)
Метаподаци
Приказ свих података о документуАпстракт
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 re...tained 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.
Извор:
Scientific Reports, 2017, 7Издавач:
- Nature Publishing Group, London
Финансирање / пројекти:
- Молекуларне особине и модификације неких респираторних и нутритивних алергена (RS-MESTD-Basic Research (BR or ON)-172024)
- Reinforcement of the Faculty of Chemistry, University of Belgrade, towards becoming a Center of Excellence in the region of WB for Molecular Biotechnology and Food research (EU-FP7-256716)
- EAACI
- Magnus Bergvall Foundation
- Hesselman Foundation
- Swedish Research Council
- Konsul ThC Bergh Foundation
- Swedish Cancer and Allergy Foundation
- King Gustaf V 80th Birthday Foundation
- Swedish Association for Allergology
- Stockholm County Council (ALF project)
- Swedish Asthma and Allergy Associations Research Foundation
- Swedish Heart-Lung Foundation
Напомена:
- Supplementary material: http://cherry.chem.bg.ac.rs/handle/123456789/3020
DOI: 10.1038/s41598-017-05355-4
ISSN: 2045-2322
PubMed: 28701697
WoS: 000405425000005
Scopus: 2-s2.0-85024102637
Колекције
Институција/група
Hemijski fakultet / Faculty of ChemistryTY - JOUR AU - Apostolović, Danijela AU - Krstić-Ristivojević, Maja AU - Mihailović-Vesić, Jelena AU - Starkhammar, Maria AU - Ćirković-Veličković, Tanja AU - Hamsten, Carl AU - van Hage, Marianne PY - 2017 UR - https://cherry.chem.bg.ac.rs/handle/123456789/2486 AB - 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. PB - Nature Publishing Group, London T2 - Scientific Reports T1 - Peptidomics of an in vitro digested alpha-Gal carrying protein revealed IgE-reactive peptides VL - 7 DO - 10.1038/s41598-017-05355-4 ER -
@article{ author = "Apostolović, Danijela and Krstić-Ristivojević, Maja and Mihailović-Vesić, Jelena and Starkhammar, Maria and Ćirković-Veličković, Tanja and Hamsten, Carl and van Hage, Marianne", year = "2017", 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.", publisher = "Nature Publishing Group, London", journal = "Scientific Reports", title = "Peptidomics of an in vitro digested alpha-Gal carrying protein revealed IgE-reactive peptides", volume = "7", doi = "10.1038/s41598-017-05355-4" }
Apostolović, D., Krstić-Ristivojević, M., Mihailović-Vesić, J., Starkhammar, M., Ćirković-Veličković, T., Hamsten, C.,& van Hage, M.. (2017). Peptidomics of an in vitro digested alpha-Gal carrying protein revealed IgE-reactive peptides. in Scientific Reports Nature Publishing Group, London., 7. https://doi.org/10.1038/s41598-017-05355-4
Apostolović D, Krstić-Ristivojević M, Mihailović-Vesić J, Starkhammar M, Ćirković-Veličković T, Hamsten C, van Hage M. Peptidomics of an in vitro digested alpha-Gal carrying protein revealed IgE-reactive peptides. in Scientific Reports. 2017;7. doi:10.1038/s41598-017-05355-4 .
Apostolović, Danijela, Krstić-Ristivojević, Maja, Mihailović-Vesić, Jelena, Starkhammar, Maria, Ćirković-Veličković, Tanja, Hamsten, Carl, van Hage, Marianne, "Peptidomics of an in vitro digested alpha-Gal carrying protein revealed IgE-reactive peptides" in Scientific Reports, 7 (2017), https://doi.org/10.1038/s41598-017-05355-4 . .