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首页> 外文期刊>Journal of Agricultural and Food Chemistry >Characterization of Allergens Isolated from the Freshwater Fish Blunt Snout Bream (Megalobrama amblycephala)
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Characterization of Allergens Isolated from the Freshwater Fish Blunt Snout Bream (Megalobrama amblycephala)

机译：从淡水鱼钝鼻鲷（Megalobrama amblycephala）分离的过敏原的表征

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Fish are an important source of dietary protein for humans throughout the world. However, they are recognized as one of the most common food allergens and pose a serious health problem in countries where fish consumption is high. Many marine fish allergens have been extensively studied, but relatively little is known about freshwater fish allergens. This study identified two main allergens from blunt snout bream (Megalobrama amblycephala), a freshwater fish widely consumed in China. Sera from 11 patients with convincing clinical history of blunt snout bream allergy were utilized in IgE immunoblot analysis to identify prominent allergens. Several blunt snout bream proteins revealed specific binding to serum IgE, with the 47 and 41 kDa proteins being the most immunodominant among them. Two-dimensional gel electrophoresis (2D SDS-PAGE) enabled resolution of the 47 and 41 kDa proteins into several protein spots with distinct isoelectric points. 2D SDS-PAGE along with IgE immunoblot analysis further confirmed the strong reactivity of these protein spots with the pooled sera from blunt snout bream-sensitive patients. Matrix-assisted laser desorption/ionizatlon time-of-flight mass spectrometry (MALDI-TOF MS) analysis of the peptides generated by trypsin digestion of the different spots corresponding to the 47 and 41 kDa proteins indicated that these spots were isoforms of enolase and muscle creatine kinase, respectively. The potential allergenicity of these proteins was further verified by an bioinformatics approach using the full-length and 80 amino acid sliding window FASTA searches, which revealed a significant amino acid sequence homology between blunt snout bream allergens and several known inhaled and crustacean allergens.
机译：鱼是全世界人类饮食蛋白质的重要​​来源。但是，它们被公认为是最常见的食物过敏原，在鱼类消费量高的国家中构成严重的健康问题。许多海水鱼类过敏原已被广泛研究，但对淡水鱼类过敏原知之甚少。这项研究从钝嘴鲷（Megalobrama amblycephala）中发现了两种主要的过敏原，钝嘴鲷在中国被广泛食用。来自11名具有令人信服的钝嘴鲷过敏临床病史的患者的血清用于IgE免疫印迹分析，以鉴定突出的过敏原。几种钝嘴鼻鲷蛋白显示出与血清IgE的特异性结合，其中47和41 kDa蛋白是其中免疫力最强的蛋白。二维凝胶电泳（2D SDS-PAGE）可以将47 kDa和41 kDa的蛋白质拆分为具有不同等电点的几个蛋白质斑点。二维SDS-PAGE和IgE免疫印迹分析进一步证实了这些蛋白斑点与钝嘴鼻鲷敏感患者的合并血清具有强反应性。基质辅助激光解吸/电离飞行时间质谱（MALDI-TOF MS）分析通过胰蛋白酶消化对应于47和41 kDa蛋白的不同斑点而产生的肽，表明这些斑点是烯醇化酶和肌肉的同工型肌酸激酶。这些蛋白质的潜在变应原性已通过生物信息学方法使用全长和80个氨基酸的滑动窗口FASTA搜索进一步验证，该结果揭示了钝嘴鼻鲷变应原与几种已知的吸入和甲壳类变应原之间存在显着的氨基酸序列同源性。

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