小麦主要过敏原CM16线性B细胞表位的预测及初步鉴定

Predicting and Identifying Linear B-Cell Epitopes of Wheat Allergen CM16

目的:探究小麦主要过敏原α-淀粉酶/胰蛋白酶抑制剂CM16的线性B细胞表位。方法:通过提取小麦蛋白粗提物,进行体外模拟胃肠道消化,发现小麦过敏原中具有消化抗性的α-淀粉酶/胰蛋白酶抑制剂CM16的片段;进一步通过生物信息学方法分析CM16的一级氨基酸序列、二级结构,并通过同源建模确定CM16的三级结构。结果:利用生物信息学法预测出CM16的4 条线性B细胞表位后,结合质谱得到的抗消化肽段信息进行比对分析,发现其中2 条肽段存在部分重合,也证明了生物信息学分析鉴定过敏原线性B细胞表位方法的可行性和应用性。结论:通过生物信息学预测小麦过敏原CM16线性B细胞表位有助于进一步认识小麦过敏原,对小麦过敏原的识别和检测具有重要的参考和借鉴作用。

Objective: To explore the feasibility and applicability of bioinformatics analysis in the identification of allergen linear B-cell epitopes. Methods: Fragments of the α-amylase/trypsin inhibitor wheat allergenic protein CM16 with digestive resistance were obtained by subjecting crude wheat protein extract (WPE) to simulated gastrointestinal digestion in vitro. The amino acid sequence and secondary structure of CM16 were analyzed by bioinformatics method, and its tertiary structure was determined by homology modeling. Results: Four linear B-cell epitopes of CM16 were predicted and compared with the digestion-resistant peptides identified by mass spectrometry (MS). It was found that two of the peptides partially overlapped, proving the feasibility and applicability of bioinformatics for identifying allergen linear B cell epitopes. Conclusion: The prediction of the linear B-cell epitopes of wheat allergen CM16 by bioinformatics will help us to gain further understanding of wheat allergens and more broadly provide an important basis for the identification and detection of wheat allergens.