Molecular Cloning and Allergenicity of Pen j 1, a Major Allergen of Kuruma Prawn, Penaeus japonicus
Molecular Cloning and Allergenicity of Pen j 1, a Major Allergen of Kuruma Prawn, Penaeus japonicus
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DOI：
10.1271/bbb.80751
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发表时间：
2009-04
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影响因子：
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通讯作者：
Ayumi Kunimoto;Takako Sisino;K. Sakai;Tomoaki Matsumoto;Kyoko Takahashi;H. Yamashita;M. Hiemori;H. Tsuji;M. Kimoto
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文献类型：
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作者：
Ayumi Kunimoto;Takako Sisino;K. Sakai;Tomoaki Matsumoto;Kyoko Takahashi;H. Yamashita;M. Hiemori;H. Tsuji;M. Kimoto
原肌球蛋白已被确定为甲壳类动物的一种常见过敏原，但其过敏性尚不清楚。在本研究中，我们从日本对虾中分离出一种变应原Pen j 1，并测定了其N端氨基酸序列。用5‘和3’端快速扩增技术(RACE)克隆了该变应原基因，编码284个氨基酸残基的蛋白质。序列分析首次表明，成熟的原肌球蛋白是由9个氨基酸残基组成的前导肽消除形成的。为明确虾过敏患者血清中免疫球蛋白E抗体的结合部位，在大肠杆菌中以谷胱甘肽S转移酶融合蛋白的形式表达了多种重组多肽，并对其与免疫球蛋白E抗体的反应性进行了检测。发现IgE结合表位存在于变应原的整个序列中，血清中的IgE抗体强烈识别其C末端区域。
Tropomyosins have been identified as a common allergen in crustaceans, but their allergenicity is not well understood. In the present study, we isolated an allergen, Pen j 1, a tropomyosin from kuruma prawn Penaeus japonicus, and determined its N-terminal amino acid sequence. The cDNA encoding the allergen was cloned by 5′- and 3′-rapid amplification of cDNA ends (RACE), and was found to code for a protein which consists of 284 amino acid residues. Sequencing analyses indicated for the first time that mature tropomyosin is formed by the elimination of a leader peptide of nine amino acid residues. To elucidate the binding sites of IgE antibodies in the sera of shrimp-sensitive patients, various recombinant peptides were expressed in Escherichia coli as fusion proteins with glutathione S-transferase (GST), and the examined with regard to reactivity with IgE antibodies. The IgE-binding epitopes were found to locate over the whole sequence of the allergen, and the IgE antibodies in the sera were found to recognize strongly its C-terminal region.