College of Environment and Public Health, Xiamen Huaxia University, 288 Tianma Road, Xiamen, Fujian 361024, China; College of Food and Biological Engineering, Jimei University, 43 Yindou Road, Xiamen, Fujian 361021, China.
College of Food and Biological Engineering, Jimei University, 43 Yindou Road, Xiamen, Fujian 361021, China.
Food Chem. 2020 Jul 1;317:126422. doi: 10.1016/j.foodchem.2020.126422. Epub 2020 Feb 17.
Myosin light chain isoform 1 (MLC1) is reported to be a novel allergen in crayfish (Procambarus clarkii). However, little information is available about its allergic epitopes. In this study, recombinant crayfish MLC1 (rMLC1) was expressed and confirmed by mass spectrometry. Circular dichroic analysis and serological test were performed for the measuring of structural and immunological properties of rMLC1. Specific-protein-A-enriched IgG raised in rabbits against purified rMLC1 was used to screen a phage display random peptide library. Nine MLC1 mimotope clones were identified among 16 random clones after biopanning. Five conformational epitopes were identified with the program LocaPep, and mapped into 3 epitope regions at the antibody-binding interface of MLC1. MLC1 of crayfish showed high primary and secondary structure identity to MLC of other allergenic species, its epitopes were located in the structure conserved regions, and its cross-reactivity among related species was indicated by immunological assays.
肌球蛋白轻链同工型 1（MLC1）被报道为小龙虾（Procambarus clarkii）中的一种新型过敏原。然而，有关其过敏原表位的信息很少。在这项研究中，重组小龙虾 MLC1（rMLC1）通过质谱法进行了表达和确认。圆二色性分析和血清学试验用于测量 rMLC1 的结构和免疫学特性。针对纯化的 rMLC1 用兔特异性蛋白 A 富集 IgG 筛选噬菌体展示随机肽文库。在生物淘选后，从 16 个随机克隆中鉴定出 9 个 MLC1 模拟克隆。使用程序 LocaPep 鉴定了 5 个构象表位，并将其映射到 MLC1 抗体结合界面的 3 个表位区域。与其他过敏原物种相比，小龙虾的 MLC1 具有高度的一级和二级结构同源性，其表位位于结构保守区域，免疫测定表明其在相关物种之间具有交叉反应性。
