Matsumoto R, Fujino K, Nagata Y, Hashiguchi S, Ito Y, Aihara Y, Takahashi Y, Maeda K, Sugimura K
Department of Bioengineering, Faculty of Engineering, Kagoshima University, Kagoshima, Japan.
Allergy. 2004 May;59(5):533-8. doi: 10.1046/j.1398-9995.2003.00412.x.
Using the sera from buckwheat (BW)-allergic patients, several putative causative molecules were reported. However, few molecules were determined on the molecular structure. We demonstrated in 2000 that the major allergen with 24 kDa (BW24KD) is a legumin-like storage protein.
The aim of this study was to isolate and characterize further a major allergen with 10 kDa by molecular cloning.
Buckwheat allergens were identified by immunoblotting analysis using sera from 14 allergic and two nonallergic individuals. We identified a protein with 10 kDa (BW10KD) that reacted with immunoglobulin E (IgE) more strongly than with IgG and IgA in 57% of the allergic patients but not with IgE in nonallergic individuals. Analyses were performed by N-terminal amino acid sequencing and molecular cloning. Physiological significance was assessed by an immunoblotting experiment showing that the reactivity of an allergic patient's serum IgE to BW10KD was competitively inhibited by natural BW extracts.
Molecular cloning experiments indicated that BW10KD as a BW allergen was a member of the 2S-albumin multigene family.
利用荞麦（BW）过敏患者的血清，已报道了几种可能的致病分子。然而，在分子结构方面确定的分子很少。我们在2000年证明，主要的24 kDa过敏原（BW24KD）是一种豆球蛋白样贮藏蛋白。
本研究的目的是通过分子克隆进一步分离和鉴定一种主要的10 kDa过敏原。
利用14名过敏个体和2名非过敏个体的血清，通过免疫印迹分析鉴定荞麦过敏原。我们鉴定出一种10 kDa的蛋白（BW10KD），在57%的过敏患者中，它与免疫球蛋白E（IgE）的反应比与IgG和IgA的反应更强，而在非过敏个体中不与IgE反应。通过N端氨基酸测序和分子克隆进行分析。通过免疫印迹实验评估其生理意义，结果表明天然BW提取物可竞争性抑制过敏患者血清IgE与BW10KD的反应性。
分子克隆实验表明，作为BW过敏原的BW10KD是2S白蛋白多基因家族的成员。
