School of Food and Biological Engineering, Engineering Research Center of Bio-process of Ministry of Education, Key Laboratory for Agricultural Products Processing of Anhui Province, Hefei University of Technology, Hefei 230009, Anhui, PR China.
School of Food and Biological Engineering, Engineering Research Center of Bio-process of Ministry of Education, Key Laboratory for Agricultural Products Processing of Anhui Province, Hefei University of Technology, Hefei 230009, Anhui, PR China.
Int J Biol Macromol. 2024 Nov;280(Pt 4):135997. doi: 10.1016/j.ijbiomac.2024.135997. Epub 2024 Sep 28.
This study examined two oleosins of 17 kDa and 15 kDa isolated from Yuzhi white sesame seeds through oil body extraction. The allergens were identified as oleosin H1 (Ses i 4) and oleosin L (Ses i 5) using SDS-PAGE, dot blot analysis, and LC-MS/MS. PCR analysis revealed high sequence homology for the oleosin proteins in the sesame seeds. Utilizing AlphaFold2, bioinformatics tools, and protein-protein docking, the structure and function of these oleosins were analyzed. Ten potential B cell epitope peptides were predicted and mapped onto the α-helix and random coil-dominated oleosome membrane conformation. IgE binding simulations identified key epitopes, B3 (FLTSGAFGL) and B4 (KRGVQEGTLY) for oleosin H1, and B8 (GGFGVAALSV) and B9 (DQLESAKTKL) for oleosin L. Mutational analysis highlighted Glu135, Phe102, Tyr128, Tyr139, Gly136, and Gly132 in oleosin H1, and Leu120, Lys119, and Leu113 in oleosin L as critical residues for binding stability, providing insights into the sensitization mechanism of these epitopes. The integration of bioinformatics and immunoinformatics in this study has contributed to a deeper understanding of the allergy properties of sesame oleosins.
本研究通过油体提取，从遇芝白芝麻中分离出两种 17kDa 和 15kDa 的油蛋白。通过 SDS-PAGE、点印迹分析和 LC-MS/MS，鉴定过敏原分别为油蛋白 H1（Ses i 4）和油蛋白 L（Ses i 5）。PCR 分析显示芝麻种子中油蛋白具有高度的序列同源性。利用 AlphaFold2、生物信息学工具和蛋白质-蛋白质对接，分析了这些油蛋白的结构和功能。预测并绘制了 10 个潜在的 B 细胞表位肽到α-螺旋和随机卷曲为主导的油体膜构象上。IgE 结合模拟确定了关键表位 B3（FLTSGAFGL）和 B4（KRGVQEGTLY）针对油蛋白 H1，以及 B8（GGFGVAALSV）和 B9（DQLESAKTKL）针对油蛋白 L。突变分析突出了油蛋白 H1 中的Glu135、Phe102、Tyr128、Tyr139、Gly136 和 Gly132，以及油蛋白 L 中的 Leu120、Lys119 和 Leu113 是结合稳定性的关键残基，为这些表位的致敏机制提供了深入的了解。本研究中生物信息学和免疫信息学的整合，有助于加深对芝麻油蛋白过敏特性的理解。
