Purification, reactivity with IgE and cDNA cloning of parvalbumin as the major allergen of mackerels
Purification, reactivity with IgE and cDNA cloning of parvalbumin as the major allergen of mackerels
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DOI：
10.1016/s0278-6915(03)00074-7
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发表时间：
2003-08-01
影响因子：
4.3
通讯作者：
Shiomi, K
中科院分区：
文献类型：
--
作者：
Hamada, Y;Tanaka, H;Shiomi, K
在日本，有三种鲱鱼(Scomber Japan onicus、S.australasicus和S.scombrus)被广泛食用，并被认为是最常发生的由IgE介导的鱼类过敏事件。在这项研究中，小白蛋白可能是主要过敏原的候选者。经Sephadex G-75凝胶过滤和TSKGelods-120T反相高效液相色谱分离，从三种鲱鱼的白色肌肉中分离纯化得到一种新的多糖组分。三个种的纯化制剂均有一条约11 kDa的单一条带，对其部分氨基酸序列进行分析，明确为细小球蛋白。在酶联免疫吸附试验中，来自鱼过敏患者的五份血清中有四份与所有纯化的小白蛋白发生反应，表明小白蛋白是与鳕鱼相同的主要过敏原。通过酶联免疫吸附抑制实验，还建立了鲱鱼细小球蛋白之间的抗原交叉反应。从日本血吸虫白色肌肉中构建了cDNA文库，并克隆了编码小白蛋白的基因。由核苷酸序列翻译而来的氨基酸序列表明，日本血吸虫小白蛋白由108个残基组成，是P型小白蛋白的一员。(C)2003爱思唯尔科学有限公司。保留所有权利。
Three species of mackerels (Scomber japonicus, S. australasicus and S. scombrus) are widely consumed and considered to be most frequently involved in incidents of IgE-mediated fish allergy in Japan. In this study, parvalbumin, a possible candidate for the major allergen. was purified from the white muscle of three species of mackerels by gel filtration on Sephadex G-75 and reverse-phase HPLC on TSKgel ODS-120T. All the purified preparations from three species gave a single band of about 11 kDa and were clearly identified as parvalbumins by analyses of their partial amino acid sequences. In ELISA experiments, four of five sera from fish-allergic patients reacted to all the purified parvalbumins, demonstrating that parvalbumin is the major allergen in common with the mackerels. Antigenic cross-reactivity among the mackerel parvalbumins was also established by ELISA inhibition experiments. A cDNA library was constructed from the white muscle of S. japonicus and the cDNA encoding parvalbumin was cloned. The amino acid sequence translated from the nucleotide sequence revealed that the S. japonicus parvalbumin is composed of 108 residues, being a member of P-type parvalbumins. (C) 2003 Elsevier Science Ltd. All rights reserved.