Identification of the major allergen of Macrobrachium rosenbergii (giant freshwater prawn)

Zailatul Hani Mohamad Yadzir; Rosmilah Misnan; Noormalin Abdullah; Faizal Bakhtiar; Masita Arip; Shahnaz Murad

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Identification of the major allergen of Macrobrachium rosenbergii (giant freshwater prawn)

机译：罗氏沼虾（巨型淡水虾）主要过敏原的鉴定

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Objective: To characterize the major allergens of Macrobrachium rosenbergii (giant freshwater prawn). Methods: Raw and cooked extracts of the giant freshwater prawn were prepared. The IgE reactivity pattern was identified by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting technique with the sera of 20 skin prick test (SPT) positive patients. The major allergen identified was then characterized using the proteomics approach involving a combination of two-dimensional (2-DE) electrophoresis, mass spectrometry and bioinformatics tools. Results: SDS-PAGE of the raw extract showed 23 protein bands (15-250 kDa) but those ranging from 40 to 100 kDa were not found in the cooked extract. From immunoblotting experiments, raw and cooked extracts demonstrated 11 and 5 IgE-binding proteins, respectively, with a molecular mass ranging from 15 to 155 kDa. A heat-resistant 36 kDa protein was identified as the major allergen of both extracts. In addition, a 42 kDa heat-sensitive protein was shown to be a major allergen of the raw extract. The 2-DE gel fractionated the prawn proteins to more than 50 different protein spots. Of these, 10 spots showed specific IgE reactivity with patients’ sera. Matrix assisted laser desorption/ionization-time of flight (MALDI-TOF) analysis led to identification of 2 important allergens, tropomyosin and arginine kinase. Conclusions: It can be concluded that the availability of such allergens would help in component-based diagnosis and therapy of prawn allergies.

机译：目的：鉴定罗氏沼虾（巨型淡水虾）的主要过敏原。方法：制备生和熟的巨型淡水虾提取物。通过十二烷基硫酸钠聚丙烯酰胺凝胶电泳（SDS-PAGE）和免疫印迹技术对20名皮肤点刺试验（SPT）阳性患者的血清进行鉴定，确定IgE反应性模式。然后使用蛋白质组学方法（包括二维（2-DE）电泳，质谱和生物信息学工具的组合）对鉴定出的主要过敏原进行表征。结果：原始提取物的SDS-PAGE显示23条蛋白带（15-250 kDa），但煮熟的提取物中未发现40至100 kDa的蛋白带。根据免疫印迹实验，生提取物和煮熟提取物分别显示出11种和5种IgE结合蛋白，分子量范围为15至155 kDa。两种提取物的主要过敏原均是耐热的36 kDa蛋白。此外，一种42 kDa的热敏蛋白被证明是原料提取物中的主要过敏原。 2-DE凝胶将虾蛋白分离成50多个不同的蛋白斑点。其中有10个斑点显示出与患者血清的特异性IgE反应性。基质辅助激光解吸/电离飞行时间（MALDI-TOF）分析导致鉴定出两种重要的过敏原，原肌球蛋白和精氨酸激酶。结论：可以得出结论，此类过敏原的可用性将有助于对虾过敏的基于成分的诊断和治疗。

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《亚太热带生物医学杂志（英文版）》 |2012年第1期|50-54|共5页
作者
Zailatul Hani Mohamad Yadzir; Rosmilah Misnan; Noormalin Abdullah; Faizal Bakhtiar; Masita Arip; Shahnaz Murad;
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Allergy and Immunology Research Centre, Institute for Medical Research, Jalan Pahang, 50588 Kuala Lumpur, Malaysia;

Department of Biology, Faculty of Science and Mathematics, Universiti Pendidikan Sultan Idris, 35900 Tanjong Malim, Perak, Malaysia;

Allergy and Immunology Research Centre, Institute for Medical Research, Jalan Pahang, 50588 Kuala Lumpur, Malaysia;

Allergy and Immunology Research Centre, Institute for Medical Research, Jalan Pahang, 50588 Kuala Lumpur, Malaysia;

Allergy and Immunology Research Centre, Institute for Medical Research, Jalan Pahang, 50588 Kuala Lumpur, Malaysia;

Allergy and Immunology Research Centre, Institute for Medical Research, Jalan Pahang, 50588 Kuala Lumpur, Malaysia;

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Identification of the major allergen of Macrobrachium rosenbergii (giant freshwater prawn)

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