Đukić Teodora, Smiljanić Katarina, Mihailović Jelena, Prodić Ivana, Apostolović Danijela, Liu Shu-Hua, Epstein Michelle M, van Hage Marianne, Stanić-Vučinić Dragana, Ćirković Veličković Tanja
University of Belgrade-Faculty of Chemistry, Department of Biochemistry and Centre of Excellence for Molecular Food Sciences, Studentski Trg 12-16, 11000 Belgrade, Serbia.
University of Belgrade-Faculty of Chemistry, Innovation Center Ltd., Studentski Trg 12-16, 11000 Belgrade, Serbia.
Foods. 2022 Dec 9;11(24):3993. doi: 10.3390/foods11243993.
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.
翻译后修饰（PTMs）是发生在蛋白质氨基酸侧链上的共价变化，但在蛋白质结构中却被忽视了其结构和功能方面。目的是使用开放式PTM搜索来检测烘焙后PTM谱的差异。我们进行了一项自下而上的蛋白质组学研究，以调查花生烘焙对易溶性过敏原及其PTM谱的影响。通过使用一系列PTM特异性抗体进行蛋白质印迹法，证实了某些修饰的蛋白质组PTM谱。除了诱导蛋白质聚集和变性外，烘焙还可能促进其PTM模式和相对谱的变化。我们已经表明，就修饰的多样性和程度而言，Ara h 1是两个样品中修饰最多的主要过敏原。最常见的PTM是甲硫氨酸氧化，尤其是在烘焙样品中。在烘焙样品中独特发现的PTM是羟基化（Trp）、甲酰化（Arg/Lys）以及氧化或羟基化（Asn）。通过酶联免疫吸附测定法（ELISA）检测，生花生提取物和烘焙花生提取物在与花生致敏个体血清中IgE的结合方面没有差异。这项研究有助于更好地理解烘焙如何影响主要花生过敏原的PTM谱，并为进一步探索PTM提供了良好的基础。
