Analysis of the Allergenic Epitopes of Tropomyosin from Mud Crab Using Phage Display and Site-Directed Mutagenesis,Journal of Agricultural and Food Chemistry

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Analysis of the Allergenic Epitopes of Tropomyosin from Mud Crab Using Phage Display and Site-Directed Mutagenesis
Journal of Agricultural and Food Chemistry ( IF 5.7 ) Pub Date : 2018-08-14 00:00:00 , DOI: 10.1021/acs.jafc.8b03466
Guang-Yu Liu ₁ , Xue-Jiao Mei ₁ , Meng-Jun Hu ₁ , Yang Yang ₁ , Meng Liu ₁ , Meng-Si Li ₁ , Ming-Li Zhang ₂ , Min-Jie Cao ₁ , Guang-Ming Liu ₁

Affiliation

Mud crab (Scylla serrata), which is widely consumed, can cause severe allergic symptoms. Eight linear epitopes and seven conformational epitopes of tropomyosin (TM) from S. serrata were identified using phage display. The conformational epitopes were formed based on the coiled-coil structure of TM. Most of the epitopes were located in the regions where primary structures were conserved among crustacean TM. Twelve synthetic peptides were designed according to the epitopes and trypsin-cutting sites of TM, among them, three synthetic peptides (including one linear epitope and two conformational epitopes) were recognized by all of the patient sera using inhibitory dot blotting. A triple-variant (R90A-E164A-Y267A) was constructed based on the critical amino acids of the TM epitope. The IgE-binding activity of the triple-variant was significantly reduced compared with that of native TM. The results of phage display and site-directed mutagenesis offered new information regarding conformational epitopes of TM.

中文翻译：

利用噬菌体展示和定点诱变技术分析来自泥蟹的原肌球蛋白的变应性抗原决定簇

被广泛食用的泥蟹（Scyla serrata）会引起严重的过敏症状。八个线性表位和从原肌球蛋白（TM）的7个构象表位S.锯缘青蟹使用噬菌体展示进行鉴定。基于TM的卷曲螺旋结构形成构象表位。大多数表位位于甲壳动物TM中保守一级结构的区域。根据TM的表位和胰蛋白酶切割位点设计了十二种合成肽，其中，通过抑制性斑点印迹法，所有患者血清均识别出三种合成肽（包括一个线性表位和两个构象性表位）。基于TM表位的关键氨基酸构建了三变体（R90A-E164A-Y267A）。与天然TM相比，三变体的IgE结合活性显着降低。噬菌体展示和定点诱变的结果提供了有关TM构象表位的新信息。

更新日期：2018-08-14

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