Liu Qing, Sui Zengying, Feng Nuan, Huang Yuhao, Li Yonghong, Ahmed Ishfaq, Ruethers Thimo, Liang Hui, Li Zhenxing, Lopata Andreas L, Sun Lirui
Department of Nutrition and Food Hygiene, School of Public Health, Qingdao University, Qingdao 266071, China.
Department of Nutrition, Qingdao Women and Children's Hospital, Qingdao 266034, China.
J Agric Food Chem. 2024 Sep 11;72(36):20077-20090. doi: 10.1021/acs.jafc.4c03944. Epub 2024 Aug 28.
Spotted seabass () is the second largest maricultural fish species in China and is the main trigger of food-related allergic reactions. Nevertheless, studies on the allergens of are limited. This study aimed to characterize pan-allergen parvalbumin from . Two proteins of about 11 kDa were purified and confirmed as parvalbumins by mass spectrometry. The IgG- and IgE-binding activities were evaluated through an immunoblotting assay. The molecular characteristics of β-parvalbumin were investigated by combining proteomics, genomics, and immunoinformatics approaches. The results indicated that β-parvalbumin consists of 109 amino acids with a molecular weight of 11.5 kDa and is the major allergen displaying strong IgE-binding capacity. In silico analysis and a dot blotting assay confirmed seven linear B cell epitopes distributed mainly on α-helixes and the calcium-binding loops. In addition, the cross-reactivity among 26 commonly consumed fish species was analyzed. The in-house generated anti- parvalbumin polyclonal antibody recognized 100% of the 26 fish species, demonstrating cross-reactivity and better binding capacity than the anticod parvalbumin antibody. Together, this study provides an efficient protocol to characterize allergens with multiomics methods and supports parvalbumin from as a candidate for fish allergen determination and allergy diagnosis.
花鲈是中国第二大海水养殖鱼类品种，也是食物相关过敏反应的主要诱发因素。然而，关于花鲈过敏原的研究却很有限。本研究旨在鉴定花鲈中的泛过敏原小清蛋白。通过质谱法纯化出两种约11 kDa的蛋白质，并确认为小清蛋白。通过免疫印迹分析评估IgG和IgE结合活性。结合蛋白质组学、基因组学和免疫信息学方法研究β-小清蛋白的分子特征。结果表明，β-小清蛋白由109个氨基酸组成，分子量为11.5 kDa，是具有强IgE结合能力的主要过敏原。计算机分析和斑点印迹分析证实了7个线性B细胞表位，主要分布在α-螺旋和钙结合环上。此外，还分析了26种常见食用鱼类之间的交叉反应性。自制的抗花鲈小清蛋白多克隆抗体识别了26种鱼类中的100%，表明具有交叉反应性，且结合能力优于抗鳕鱼小清蛋白抗体。总之，本研究提供了一种用多组学方法鉴定过敏原的有效方案，并支持将花鲈中的小清蛋白作为鱼类过敏原测定和过敏诊断的候选物。
