Laboratory of Biochemistry and Molecular Biology, College of Life Science, Northwest A&F University, Yangling, China.
Laboratory of Biochemistry and Molecular Biology, College of Life Science, Northwest A&F University, Yangling, China.
Food Chem. 2021 Jun 1;346:128953. doi: 10.1016/j.foodchem.2020.128953. Epub 2020 Dec 26.
Tartary buckwheat is widely accepted as its nutritionalvalue. Some allergic reactions hinder its utilization. This research focused on evaluating the core epitope of 16 kDa allergen (Fag t 2) in tartary buckwheat. Six B- and seven T cell epitopes of Fag t 2 were predicted, and six B cell epitope-mutants were expressed in Pichia pastoris. Bioinformatics analysis and SDS-PAGE demonstrated that the molecular weight, isoelectric point and spatial structures of six mutant allergens were similar with Fag t 2, with the same signal peptide sequences and α-amylase inhibitor domain. There was no significant change in mutants' spatial conformation confirmed by Circular Dichroism. The position K132N and peptides at 108-117 and 132-141 were the core B- and T cell epitopes of Fag t 2 confirmed by competitive inhibition ELISA and dot blot. This result was of great significance on the study of allergen epitopes in prevention and treatment of hypersensitivity.
苦荞被广泛认可其营养价值。一些过敏反应阻碍了它的利用。本研究聚焦于评估苦荞中 16 kDa 过敏原（Fag t 2）的核心表位。预测了 Fag t 2 的 6 个 B 细胞和 7 个 T 细胞表位，并在毕赤酵母中表达了 6 个 B 细胞表位突变体。生物信息学分析和 SDS-PAGE 表明，6 种突变过敏原的分子量、等电点和空间结构与 Fag t 2 相似，具有相同的信号肽序列和 α-淀粉酶抑制剂结构域。圆二色性分析表明突变体的空间构象没有明显变化。通过竞争抑制 ELISA 和斑点印迹法证实，位置 K132N 和肽 108-117 及 132-141 是 Fag t 2 的核心 B 细胞和 T 细胞表位。这一结果对于研究过敏原表位在预防和治疗过敏中的作用具有重要意义。
