Proteomic Profiling of Major Peanut Allergens and Their Post-Translational Modifications Affected by Roasting
Teodora Đukić,
Katarina Smiljanić,
Jelena Mihailović,
Ivana Prodić,
Danijela Apostolović,
Shu-Hua Liu,
Michelle M. Epstein,
Marianne van Hage,
Dragana Stanić-Vučinić,
Tanja Ćirković Veličković
Affiliations
Teodora Đukić
University of Belgrade—Faculty of Chemistry, Department of Biochemistry and Centre of Excellence for Molecular Food Sciences, Studentski Trg 12–16, 11000 Belgrade, Serbia
Katarina Smiljanić
University of Belgrade—Faculty of Chemistry, Department of Biochemistry and Centre of Excellence for Molecular Food Sciences, Studentski Trg 12–16, 11000 Belgrade, Serbia
Jelena Mihailović
University of Belgrade—Faculty of Chemistry, Department of Biochemistry and Centre of Excellence for Molecular Food Sciences, Studentski Trg 12–16, 11000 Belgrade, Serbia
Ivana Prodić
University of Belgrade—Faculty of Chemistry, Innovation Center Ltd., Studentski Trg 12-16, 11000 Belgrade, Serbia
Danijela Apostolović
Department of Medicine Solna, Division of Immunology and Allergy, Karolinska Institutet and Karolinska University Hospital, SE-171 76 Stockholm, Sweden
Shu-Hua Liu
Medical University of Vienna Department of Dermatology, Experimental Allergy Laboratory, Waehringer Guertel 18–20, 1090 Vienna, Austria
Michelle M. Epstein
Medical University of Vienna Department of Dermatology, Experimental Allergy Laboratory, Waehringer Guertel 18–20, 1090 Vienna, Austria
Marianne van Hage
Department of Medicine Solna, Division of Immunology and Allergy, Karolinska Institutet and Karolinska University Hospital, SE-171 76 Stockholm, Sweden
Dragana Stanić-Vučinić
University of Belgrade—Faculty of Chemistry, Department of Biochemistry and Centre of Excellence for Molecular Food Sciences, Studentski Trg 12–16, 11000 Belgrade, Serbia
Tanja Ćirković Veličković
University of Belgrade—Faculty of Chemistry, Department of Biochemistry and Centre of Excellence for Molecular Food Sciences, Studentski Trg 12–16, 11000 Belgrade, Serbia
Post-translational modifications (PTMs) are covalent changes occurring on amino acid side chains of proteins and yet are neglected structural and functional aspects of protein architecture. The objective was to detect differences in PTM profiles that take place after roasting using open PTM search. We conducted a bottom-up proteomic study to investigate the impact of peanut roasting on readily soluble allergens and their PTM profiles. Proteomic PTM profiling of certain modifications was confirmed by Western blotting with a series of PTM-specific antibodies. In addition to inducing protein aggregation and denaturation, roasting may facilitate change in their PTM pattern and relative profiling. We have shown that Ara h 1 is the most modified major allergen in both samples in terms of modification versatility and extent. The most frequent PTM was methionine oxidation, especially in roasted samples. PTMs uniquely found in roasted samples were hydroxylation (Trp), formylation (Arg/Lys), and oxidation or hydroxylation (Asn). Raw and roasted peanut extracts did not differ in the binding of IgE from the serum of peanut-sensitised individuals done by ELISA. This study provides a better understanding of how roasting impacts the PTM profile of major peanut allergens and provides a good foundation for further exploration of PTMs.
