Molecular characterization of a 10-kDa buckwheat molecule reactive to allergic patients' IgE
Molecular characterization of a 10-kDa buckwheat molecule reactive to allergic patients' IgE
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DOI：
10.1046/j.1398-9995.2003.00412.x
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发表时间：
2004-05-01
期刊：
影响因子：
12.4
通讯作者：
Sugimura, K
中科院分区：
文献类型：
--
作者：
Matsumoto, R;Fujino, K;Sugimura, K
背景：利用荞麦（BW）过敏患者的血清，报道了几种假定的致病分子。然而，很少有分子被确定的分子结构。我们在2000年证明，24 kDa（BW 24 KD）的主要过敏原是一个豆类蛋白样storage protein.Objective：本研究的目的是分离和表征进一步的主要过敏原与10 kDa的分子cloning.Methods和结果：荞麦过敏原的免疫印迹分析鉴定14过敏和两个非过敏个体的血清。我们确定了一种10 kDa（BW 10 KD）的蛋白质，在57%的过敏性患者中与免疫球蛋白E（IgE）的反应比与IgG和伊加的反应更强烈，但在非过敏性个体中不与IgE反应。通过N-末端氨基酸测序和分子克隆进行分析。生理意义进行了评估的免疫印迹实验表明，过敏患者的血清IgE的反应性BW 10 KD的竞争性抑制天然BW extract.Conclusion：分子克隆实验表明，BW 10 KD作为BW过敏原是2S-白蛋白多基因家族的成员。
Background: 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.Objective: The aim of this study was to isolate and characterize further a major allergen with 10 kDa by molecular cloning.Methods and results: 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.Conclusion: Molecular cloning experiments indicated that BW10KD as a BW allergen was a member of the 2S-albumin multigene family.