| 桂沄坤,王锋,陈淑情,等.鹰嘴桃过敏原的鉴定及其生物学特性分析[J].广东农业科学,2024,(8-10):- |
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鹰嘴桃过敏原的鉴定及其生物学特性分析 |
| Identification and Analysis of Biological Characteristics of Olecranon Peach Allergens |
| 投稿时间：2024-05-11 修订日期：2024-07-13 |
| DOI： |
| 中文关键词: 鹰嘴桃 过敏原蛋白 蛋白质谱 生物学特性 蛋白结构 抗原表位 |
| 英文关键词: olecranon peaches allergenic proteins identification of allergens biological characteristics protein structures antigenic epitopes |
| 基金项目:国家自然科学基金项目(32202175)；广东省基础与应用基础研究基金项目(2021A1515110608)； “十四五”广东省农业科技创新十大主攻方向“揭榜挂帅”项目(2024KJ15)；广东省普通高校特色创新类项目(2023KTSCX049)；广州市科技计划项目（2024A04J5001）；广东省重点建设学科科研能力提升项目(2022ZDJS021) ；仲恺农业工程学院大学生创新创业训练计划项目(S202311347049) |
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| 中文摘要: |
| 【目的】鹰嘴桃作为岭南地域的特色水果，因为其良好的风味和品质被评为“岭南十大佳果”，但未见关于其过敏原的研究，通过分析其过敏原蛋白的抗原表位，为过敏原重组抗原的制备提供研究基础，也为阐述食品加工过程中鹰嘴桃过敏原的特性变化和致敏特性提供研究依据。【方法】本文提取并鉴定了鹰嘴桃中存在的过敏原，并利用多种生物学方法预测分析了过敏原的生物学特性。用磷酸缓冲液提取鹰嘴桃冻干粉中的粗蛋白，采用SDS-PAGE蛋白电泳方法鉴定并联合蛋白质谱方法分析了粗蛋白中存在的过敏原，通过蛋白数据库UniProt进行筛选比对，并采用生物信息学方法分析致敏蛋白的理化性质、空间结构和抗原表位。【结果】在鹰嘴桃中鉴定出 7 种致敏蛋白（A0A251RBV3、P86888、M5X697、M5WV03、M5WTQ8、Q2I6V8、Q9LED1），主要归属于病程相关蛋白（Pru p 1）、类甜蛋白（Pru p 2）、非特异性脂质转移蛋白家族（Pru p 3）和赤霉素调节蛋白（Pru p 7）四类过敏原蛋白。7种鹰嘴桃过敏原蛋白具有较高稳定性，脂肪族氨基酸指数为 29.3~81.54。M5WTQ8 和 Q2I6V8 过敏原蛋白为酸性蛋白，其余过敏原蛋白为碱性蛋白。除 Q9LED1 蛋白外，其余过敏原蛋白均为亲水蛋白。筛选抗原表位、亲水性和柔韧性大于 0 且表面可及性大于 1 的区域，并结合蛋白的二级和三级结构分析蛋白键能较低的区域，获得鹰嘴桃过敏原的抗原表位分别为Pru p 1(EIP，GSQ，KEN，NL，KG，EIK，HPD)、Pru p 2（TGDQKPQ，SP，NQ，PPNDKPETCPPT，DDKSS，RP）、Pru p 3（RT，VN）和 Pru p 7(AGY，GTYGN，LKNSKGN)。【结论】通过对过敏原蛋白的结构、亲水性、表面可及性、柔韧性和抗原指数进行分析，获得过敏原蛋白的多个抗原表位。 |
| 英文摘要: |
| 【Objective】The olecranon peach is known as the characteristic fruit of Lingnan region because of its good flavor and quality, and it is named as "The top ten best fruits in Lingnan". However, less studies on its allergens have been reported. By analyzing the epitope of the allergen protein, it provides the research basis for the preparation of the recombinant antigen of the allergen, and also provides the research basis for explaining the characteristic changes and sensitization characteristics of the allergen in the process of food processing.【Method】Allergens in olecranon peach were extracted and identified, and the biological characteristics of the allergens were analyzed using a variety of biological methods. Phosphate buffer saline was used to extract the crude protein from the lyophilized powder of olecranon peach, and the allergens in the crude protein were identified by SDS-PAGE protein electrophoresis and analyzed by mass spectrometry.. The allergens were screened and evaluated by the protein database of UniProt, and the physicochemical properties, spatial structure and antigenic epitopes of the allergens were analyzed by bioinformatics methods.【Result】The results showed that seven kinds of allergenic proteins including A0A251RBV3, P86888, M5X697, M5WV03, M5WTQ8, Q2I6V8, and Q9LED1 were identified from olecranon peach, Which belonged to the four classes of allergenic proteins pathogenesis-related proteins (Pru p 1), thaumatin-like proteins (Pru p 2), non-specific Lipid Transfer protein (Pru p 3) and gibberellin-regulated proteins (Pru p 7). M5WTQ8 and Q2I6V8 allergen proteins were acidic proteins, while the remaining allergen proteins were alkaline proteins. seven kinds of allergenic proteins in olecranon peach had high stability with the aliphatic index between 29.3 and 81.54. Except for Q9LED1 protein, all other allergen proteins were hydrophilic proteins. The regions with antigen epitopes, hydrophilicity and flexibility greater than 0 and surface accessibility greater than 1 are screened, and regions with low protein bonding energy are analyzed by binding the secondary and tertiary structures of proteins to obtain the sequence of B-cell epitope-binding amino acids: Pru p1 (EIP, GSQ, KEN, NL, KG, EIK, HPD), Pru p2 (TGDQKPQ, SP, NQ, PPNDKPETCPPT, DDKSS, RP), Pru p3 (RT, VN) and Pru p7 (AGY, GTYGN, LKNSKGN).【Conclusion】By analyzing the structure, hydrophilicity, surface accessibility, flexibility and antigen index of olecranon peach, multiple epitopes of 7 allergens were obtained. |
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