Contribution of Structural Reversibility to the Heat Stability of the Tropomyosin Shrimp Allergen
Contribution of Structural Reversibility to the Heat Stability of the Tropomyosin Shrimp Allergen
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DOI：
10.1271/bbb.120887
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发表时间：
2013-05
期刊：
影响因子：
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通讯作者：
M. Usui;A. Harada;T. Ishimaru;Emiri Sakumichi;Fumihiko Saratani;Chiho Sato-Minami;H. Azakami;T. Miyasaki;K. Hanaoka
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文献类型：
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作者：
M. Usui;A. Harada;T. Ishimaru;Emiri Sakumichi;Fumihiko Saratani;Chiho Sato-Minami;H. Azakami;T. Miyasaki;K. Hanaoka
原肌球蛋白是常见的热稳定的甲壳类过敏原。然而，它们的热稳定性及其对抗原性的影响尚未明确。本研究从未经热处理的生日本麻仁对虾中纯化原肌球蛋白。SDS-PAGE结果显示，纯化蛋白与对虾过敏患者血清中的IgE发生了约35 kDa的交叉反应，鉴定为Pen j 1。原肌球蛋白的圆二色光谱显示，原肌球蛋白具有常见的α-螺旋结构，在加热至80℃时容易崩溃。然而，加热后没有不溶性聚集体，冷却至25°C后蛋白质恢复其原始CD光谱模式。自然致敏与加热致敏小鼠的总IgG产量无显著差异。这些结果表明，热变性的Pen j1在冷却后重新折叠，并在热处理后保持其抗原性。
Tropomyosins are common heat-stable crustacean allergens. However, their heat stability and their effects on antigenicity have not been clarified. We purified tropomyosin in this study from raw kuruma prawns (Marsupenaeus japonicus) without heat processing. SDS-PAGE of the purified protein showed a band at approximately 35 kDa that cross-reacted with IgE from the serum of a shrimp-allergic patient, identifying it as Pen j 1. The circular dichroism spectrum of native Pen j 1 revealed the common α-helical structure of tropomyosins which easily collapsed upon heating to 80 °C. However, there were no insoluble aggregates after heating, and the protein regained its native CD spectral pattern after cooling to 25 °C. There was no significant difference in total IgG production between mice sensitized with native and heated Pen j 1. These results suggest that heat-denatured Pen j 1 refolded upon cooling and maintained its antigenicity following the heat treatment.