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利用患者血清和基于 MS 的蛋白质组学分析鉴定胡芦巴(Trigonella foenum-graecum)中的潜在过敏原。

Characterization of potential allergens in fenugreek (Trigonella foenum-graecum) using patient sera and MS-based proteomic analysis.

机构信息

National Veterinary Institute, Oslo, Norway.

出版信息

J Proteomics. 2010 May 7;73(7):1321-33. doi: 10.1016/j.jprot.2010.02.011. Epub 2010 Feb 26.

Abstract

BACKGROUND

Fenugreek is a legume plant used as an ingredient of curry spice. Incidents of IgE-mediated food allergy to fenugreek have been reported. Coincidence with allergy to peanut, a major food allergen, seems to be common suggesting a rather high rate of cross-reactivity.

OBJECTIVE

Characterization of fenugreek allergens using patient sera and mass spectrometry-based proteomic analysis.

METHODS

Allergenic fenugreek proteins were detected by immunoblotting, using sera from 13 patients with specific IgE to peanut and fenugreek. IgE-binding proteins were analyzed by peptide mass fingerprinting and peptide sequencing.

RESULTS

A fenugreek protein quintet in the range from 50 kDa to 66 kDa showed high IgE-affinity, the protein at 50 kDa reaching the strongest signals in all patients. Proteomic analyses allowed the classification of several fenugreek proteins to a number of allergen families. Fenugreek 7S-vicilin and 11S-legumin were partly sequenced and revealed considerable homologies to peanut Ara h 1 and Ara h 3, respectively. The presence of a fenugreek 2S albumin and pathogenesis-related (PR-10) plant pollen protein was assumed by database searching results.

CONCLUSION

In this study, individual fenugreek proteins were characterised for the first time. Observed homologies to major peanut allergens provide a molecular explanation for clinical cross-reactivity.

摘要

背景

葫芦巴是一种豆科植物,用作咖喱香料的成分。已报告了对葫芦巴的 IgE 介导的食物过敏事件。与花生(主要食物过敏原)过敏的巧合似乎很常见,这表明存在相当高的交叉反应率。

目的

使用患者血清和基于质谱的蛋白质组学分析来鉴定葫芦巴过敏原。

方法

使用 13 名对花生和葫芦巴特异性 IgE 的患者的血清,通过免疫印迹法检测致敏的葫芦巴蛋白。通过肽质量指纹图谱和肽测序分析 IgE 结合蛋白。

结果

50 kDa 至 66 kDa 范围内的葫芦巴蛋白五聚体表现出高 IgE 亲和力,所有患者中 50 kDa 的蛋白均产生最强的信号。蛋白质组学分析将几种葫芦巴蛋白分类为许多过敏原家族。葫芦巴 7S-豆球蛋白和 11S-豆球蛋白部分测序,并分别显示出与花生 Ara h 1 和 Ara h 3 的相当大的同源性。通过数据库搜索结果推测存在葫芦巴 2S 白蛋白和与发病机理相关的(PR-10)植物花粉蛋白。

结论

在这项研究中,首次对个别葫芦巴蛋白进行了表征。观察到与主要花生过敏原的同源性为临床交叉反应提供了分子解释。

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