斑节对虾（凡纳滨对虾）和澳洲对虾（宽沟对虾）主要和次要过敏原的鉴定。
Identification of Major and Minor Allergens of Black Tiger Prawn (Penaeus monodon) and King Prawn (Penaeus latisulcatus).
作者信息
Sahabudin Syuhaidah, Misnan Rosmilah, Yadzir Zailatul Hani Mohammad, Mohamad Jamaludin, Abdullah Noormalin, Bakhtiar Faizal, Murad Shahnaz
机构信息
Allergy and Immunology Research Centre, Institute for Medical Research, 50588 Kuala Lumpur, Malaysia.
出版信息
Malays J Med Sci. 2011 Jul;18(3):27-32.
BACKGROUND
Prawns and shrimp are a frequent cause of seafood allergy mediated by IgE antibodies. Penaeus monodon and Penaeus latisulcatus, commonly known as black tiger prawn and king prawn, respectively, are among the most frequently consumed prawns in Malaysia. The aim of this study was to identify the IgE-binding proteins of these 2 prawn species.
METHODS
Raw and boiled prawn extracts were prepared and then resolved by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). IgE-immunoblotting was then performed using sera from patients with positive skin prick tests to the raw prawn extracts.
RESULTS
SDS-PAGE analysis of the raw extracts of both prawn species revealed 23 protein bands; the boiled extracts yielded fewer protein bands. The bands in the range of 40 to 100 kDa were sensitive to heat and therefore were not found in the boiled extracts. Immunoblot of raw extracts of black tiger prawns and king prawns yielded 14 and 11 IgE-binding proteins, respectively, with molecular weights of between 15 and 200 kDa. Proteins at 36, 42, and 49 kDa were detected as the major allergens in both species of prawns. A protein of 75 kDa was also identified as a major allergen in black tiger prawns. Other potential allergens were also observed at various molecular masses.
CONCLUSION
Proteins of 36, 42, and 49 kDa were identified as the major allergens of both species of prawns. The 36 and 42 kDa proteins are hypothesised to be tropomyosin and arginine kinase, respectively. A high molecular weight protein of 75 kDa was found to be an additional major allergen in black tiger prawns.
背景
对虾和小虾是由IgE抗体介导的海鲜过敏的常见诱因。斑节对虾和宽沟对虾，通常分别被称为黑虎虾和明虾，是马来西亚最常食用的对虾品种。本研究的目的是鉴定这两种对虾的IgE结合蛋白。
方法
制备生的和煮熟的对虾提取物，然后通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）进行分离。然后使用对生对虾提取物皮肤点刺试验呈阳性的患者血清进行IgE免疫印迹分析。
结果
两种对虾生提取物的SDS-PAGE分析显示有23条蛋白带；煮熟的提取物产生的蛋白带较少。40至100 kDa范围内的条带对热敏感，因此在煮熟的提取物中未发现。黑虎虾和明虾生提取物的免疫印迹分别产生了14种和11种IgE结合蛋白，分子量在15至200 kDa之间。在两种对虾中均检测到36、42和49 kDa的蛋白为主要过敏原。75 kDa的蛋白也被鉴定为黑虎虾的主要过敏原。在各种分子量处还观察到其他潜在过敏原。
结论
36、42和49 kDa的蛋白被鉴定为两种对虾的主要过敏原。推测36 kDa和42 kDa的蛋白分别为原肌球蛋白和精氨酸激酶。发现75 kDa的高分子量蛋白是黑虎虾的另一种主要过敏原。
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