Comparative study of raw and thermally treated peanut major allergen post- translational modifications (PTMs)
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Introduction. Peanut allergy affects a large portion of world population causing reactions ranging
from mild to severe. Major peanut allergen IgE epitopes are well characterized but little is known
about their post-translational modifications (PTM) and how they are affected by thermal
treatment. PTM profile may differ between raw and thermally treated peanut, which could affect
its allergic potential depending on type, size and position of modifications.
Objective. Our aim was to analyse and compare PTM profiles of 4 major peanut allergens - Ara h 1,
Ara h 2, Ara h 3 and Ara h 6, as well as their amounts in raw and roasted samples using bottom-up
proteomics methods.
Methodology. Full peanut protein extracts (both thermally treated and non-treated) were digested
in gel and in solution, and analysed by a Top10 nLC-MS/MS method by LTQ Orbitrap XL (Thermo
Fisher Scientific Inc., Germany). Within the extracts major allergens - Ara h 1, Ara h 2, Ara h 3 and
Ara h 6 were identified..., label free quantified (LFQ) and searched for PTMs by Peaks X software
(Bioinformatics solutions Inc.I, Canada). Epitope sequences were acquired from the Immune
Epitope Database (IEDB www.iedb.org).
Main findings. LFQ results show that there is no significant change in the amountsof any of the
studied allergens between raw and roasted extracts.Out of the 4 allergens Ara h 6 is modified in the
highest portion, with respect to the protein size: 15% and 12% of its positions are modified in raw
and roasted sample, respectively. Total of 21 modifications were quantified between the two
preparations, with oxidation (M), methylation (K,R) and dethiomethylation affecting the largest
number of peptides.
Conclusions. Peanut allergen epitopes are indeed carriers of PTMs that differ in pattern and
quantity between treated and non-treated extracts. The in silico discovered PTMs could affect
protein digestibility and allergenicity. Further investigation is necessary in order to fully understand
the impact protein modifications could have on their allergenic potential.
Ključne reči:
mass spectrometry based bottom-up proteomics / PTMs / LFQ / relative quantification / allergens / peanutIzvor:
Book of Abstracts - V SePa Simposium: Proteomics in the analysis of food, environmental protection and medical research, Novi Sad 2019, 2019, 16/L10-Izdavač:
- Serbian Proteomic Association - SePA
Finansiranje / projekti:
- Molekularne osobine i modifikacije nekih respiratornih i nutritivnih alergena (RS-MESTD-Basic Research (BR or ON)-172024)
- Ghent University Global Campus, Belgian Special Research Fund BOF StG No. 01N01718.
- FoodEnTwin-Twinning of research activities for the frontier research in the fields of food, nutrition and environmental omics (EU-H2020-810752)
Kolekcije
Institucija/grupa
Hemijski fakultet / Faculty of ChemistryTY - CONF AU - Mihailović, Jelena AU - Prodić, Ivana AU - Smiljanić, Katarina AU - Ćirković Veličković, Tanja PY - 2019 UR - http://cherry.chem.bg.ac.rs/handle/123456789/5725 AB - Introduction. Peanut allergy affects a large portion of world population causing reactions ranging from mild to severe. Major peanut allergen IgE epitopes are well characterized but little is known about their post-translational modifications (PTM) and how they are affected by thermal treatment. PTM profile may differ between raw and thermally treated peanut, which could affect its allergic potential depending on type, size and position of modifications. Objective. Our aim was to analyse and compare PTM profiles of 4 major peanut allergens - Ara h 1, Ara h 2, Ara h 3 and Ara h 6, as well as their amounts in raw and roasted samples using bottom-up proteomics methods. Methodology. Full peanut protein extracts (both thermally treated and non-treated) were digested in gel and in solution, and analysed by a Top10 nLC-MS/MS method by LTQ Orbitrap XL (Thermo Fisher Scientific Inc., Germany). Within the extracts major allergens - Ara h 1, Ara h 2, Ara h 3 and Ara h 6 were identified, label free quantified (LFQ) and searched for PTMs by Peaks X software (Bioinformatics solutions Inc.I, Canada). Epitope sequences were acquired from the Immune Epitope Database (IEDB www.iedb.org). Main findings. LFQ results show that there is no significant change in the amountsof any of the studied allergens between raw and roasted extracts.Out of the 4 allergens Ara h 6 is modified in the highest portion, with respect to the protein size: 15% and 12% of its positions are modified in raw and roasted sample, respectively. Total of 21 modifications were quantified between the two preparations, with oxidation (M), methylation (K,R) and dethiomethylation affecting the largest number of peptides. Conclusions. Peanut allergen epitopes are indeed carriers of PTMs that differ in pattern and quantity between treated and non-treated extracts. The in silico discovered PTMs could affect protein digestibility and allergenicity. Further investigation is necessary in order to fully understand the impact protein modifications could have on their allergenic potential. PB - Serbian Proteomic Association - SePA C3 - Book of Abstracts - V SePa Simposium: Proteomics in the analysis of food, environmental protection and medical research, Novi Sad 2019 T1 - Comparative study of raw and thermally treated peanut major allergen post- translational modifications (PTMs) SP - 16/L10 UR - https://hdl.handle.net/21.15107/rcub_cherry_5725 ER -
@conference{ author = "Mihailović, Jelena and Prodić, Ivana and Smiljanić, Katarina and Ćirković Veličković, Tanja", year = "2019", abstract = "Introduction. Peanut allergy affects a large portion of world population causing reactions ranging from mild to severe. Major peanut allergen IgE epitopes are well characterized but little is known about their post-translational modifications (PTM) and how they are affected by thermal treatment. PTM profile may differ between raw and thermally treated peanut, which could affect its allergic potential depending on type, size and position of modifications. Objective. Our aim was to analyse and compare PTM profiles of 4 major peanut allergens - Ara h 1, Ara h 2, Ara h 3 and Ara h 6, as well as their amounts in raw and roasted samples using bottom-up proteomics methods. Methodology. Full peanut protein extracts (both thermally treated and non-treated) were digested in gel and in solution, and analysed by a Top10 nLC-MS/MS method by LTQ Orbitrap XL (Thermo Fisher Scientific Inc., Germany). Within the extracts major allergens - Ara h 1, Ara h 2, Ara h 3 and Ara h 6 were identified, label free quantified (LFQ) and searched for PTMs by Peaks X software (Bioinformatics solutions Inc.I, Canada). Epitope sequences were acquired from the Immune Epitope Database (IEDB www.iedb.org). Main findings. LFQ results show that there is no significant change in the amountsof any of the studied allergens between raw and roasted extracts.Out of the 4 allergens Ara h 6 is modified in the highest portion, with respect to the protein size: 15% and 12% of its positions are modified in raw and roasted sample, respectively. Total of 21 modifications were quantified between the two preparations, with oxidation (M), methylation (K,R) and dethiomethylation affecting the largest number of peptides. Conclusions. Peanut allergen epitopes are indeed carriers of PTMs that differ in pattern and quantity between treated and non-treated extracts. The in silico discovered PTMs could affect protein digestibility and allergenicity. Further investigation is necessary in order to fully understand the impact protein modifications could have on their allergenic potential.", publisher = "Serbian Proteomic Association - SePA", journal = "Book of Abstracts - V SePa Simposium: Proteomics in the analysis of food, environmental protection and medical research, Novi Sad 2019", title = "Comparative study of raw and thermally treated peanut major allergen post- translational modifications (PTMs)", pages = "16/L10", url = "https://hdl.handle.net/21.15107/rcub_cherry_5725" }
Mihailović, J., Prodić, I., Smiljanić, K.,& Ćirković Veličković, T.. (2019). Comparative study of raw and thermally treated peanut major allergen post- translational modifications (PTMs). in Book of Abstracts - V SePa Simposium: Proteomics in the analysis of food, environmental protection and medical research, Novi Sad 2019 Serbian Proteomic Association - SePA., 16/L10. https://hdl.handle.net/21.15107/rcub_cherry_5725
Mihailović J, Prodić I, Smiljanić K, Ćirković Veličković T. Comparative study of raw and thermally treated peanut major allergen post- translational modifications (PTMs). in Book of Abstracts - V SePa Simposium: Proteomics in the analysis of food, environmental protection and medical research, Novi Sad 2019. 2019;:16/L10. https://hdl.handle.net/21.15107/rcub_cherry_5725 .
Mihailović, Jelena, Prodić, Ivana, Smiljanić, Katarina, Ćirković Veličković, Tanja, "Comparative study of raw and thermally treated peanut major allergen post- translational modifications (PTMs)" in Book of Abstracts - V SePa Simposium: Proteomics in the analysis of food, environmental protection and medical research, Novi Sad 2019 (2019):16/L10, https://hdl.handle.net/21.15107/rcub_cherry_5725 .