Ogawa T, Tsuji H, Bando N, Kitamura K, Zhu Y L, Hirano H, Nishikawa K
Department of Nutrition, School of Medicine, University of Tokushima, Japan.
Biosci Biotechnol Biochem. 1993 Jun;57(6):1030-3. doi: 10.1271/bbb.57.1030.
The soybean allergenic protein, Gly m Bd 30K [Ogawa et al., J. Nutr. Sci. Vitaminol., 37, 555-565 (1991)] which is most strongly and frequently recognized by the IgE antibodies in sera of soybean-sensitive patients with atopic dermatitis, has been characterized. The allergen was isolated from the crude 7S-globulin fraction as an oligomeric form with a molecular weight of more than 3000,000 by gel-filtration chromatography. On two-dimensional gel electrophoresis, the native oligomeric allergen had an isoelectric point of about pH 4.5 and was dissociated into a monomeric form with a molecular weight of about 32,000 by the treatment with sodium dodecyl sulfate and 2-mercaptoethanol. The monomeric allergen had an N-terminal amino acid sequence and amino acid composition identical with those of the soybean seed 34-kDa oil-body-associated protein or the soybean vacuolar protein P34 with close homology to papain-like thiol proteinases [Kalinski et al., J. Biol. Chem., 267, 12068 (1992)]. The identity was further confirmed by the immunological cross-reactivity to the antibodies produced against each of the purified allargen and the 34-kDa oil-body-associated protein. By this observation, Gly m Bd 30K was shown to have about 30% sequence homology with Der pI, a house dust mite allergen that is a thiol proteinase from Dermatophagoides pteronyssius.
大豆致敏蛋白Gly m Bd 30K [小川等人，《营养科学与维生素学杂志》，37，555 - 565（1991）]已被鉴定，该蛋白在患有特应性皮炎的大豆敏感患者血清中最强烈且最频繁地被IgE抗体识别。通过凝胶过滤色谱法，从粗制的7S球蛋白级分中分离出该过敏原，其为分子量超过3000,000的寡聚体形式。在二维凝胶电泳上，天然寡聚体过敏原的等电点约为pH 4.5，用十二烷基硫酸钠和2 - 巯基乙醇处理后解离成分子量约为32,000的单体形式。单体过敏原的N端氨基酸序列和氨基酸组成与大豆种子34 kDa油体相关蛋白或与木瓜蛋白酶样巯基蛋白酶具有高度同源性的大豆液泡蛋白P34相同[卡林斯基等人，《生物化学杂志》，267，12068（1992）]。通过对针对纯化的过敏原和34 kDa油体相关蛋白产生的抗体的免疫交叉反应进一步证实了这种同一性。通过这一观察结果表明，Gly m Bd 30K与屋尘螨过敏原Der pI具有约30%的序列同源性，Der pI是一种来自粉尘螨的巯基蛋白酶。
