Institute of Plant Genetics and Biotechnology, Slovak Academy of Sciences , 950 07, Nitra, Slovakia.
J Proteome Res. 2013 Nov 1;12(11):4862-9. doi: 10.1021/pr400336f. Epub 2013 Sep 5.
The amount of clinically relevant, allergy-related proteins in wheat grain is still largely unknown. The application of proteomics may create a platform not only for identification and characterization, but also for quantitation of these proteins. The aim of this study was to evaluate the data-independent quantitative mass spectrometry (MS(E)) approach in combination with 76 wheat allergenic sequences downloaded from the AllergenOnline database ( www.allergenonline.org ) as a starting point. Alcohol soluble extracts of gliadin and glutenin proteins were analyzed. This approach has resulted in identification and quantification of 15 allergenic protein isoforms that belong to amylase/trypsin inhibitors, γ-gliadins, and high or low molecular weight glutenins. Additionally, several peptides carrying four previously discovered epitopes of γ-gliadin B precursor have been detected. These data were validated against the UniProt database, which contained 11764 Triticeae protein sequences. The identified allergens are discussed in relation to Baker's asthma, food allergy, wheat dependent exercise induced anaphylaxis, atopic dermatitis, and celiac disease (i.e., gluten-sensitive enteropathy). In summary, the results showed that the MS(E) approach is suitable for quantitative analysis and allergens profiling in wheat varieties and/or other food matrices.
小麦中与临床相关的、与过敏相关的蛋白质的含量在很大程度上仍不清楚。蛋白质组学的应用不仅可以为这些蛋白质的鉴定和特性分析创建平台，还可以进行定量分析。本研究旨在评估数据非依赖型定量质谱（MS(E)）方法，同时以从 AllergenOnline 数据库（www.allergenonline.org）下载的 76 个小麦过敏原序列作为起点。醇溶性麦醇溶蛋白和麦谷蛋白提取物进行了分析。该方法鉴定和定量了 15 种过敏原蛋白同工型，它们属于淀粉酶/胰蛋白酶抑制剂、γ-麦谷蛋白和高分子量或低分子量麦谷蛋白。此外，还检测到了几个携带γ-麦谷蛋白 B 前体四个先前发现的表位的肽段。这些数据与包含 11764 个三磷酸腺苷结合盒转运蛋白序列的 UniProt 数据库进行了验证。讨论了所鉴定的过敏原与烘焙师哮喘、食物过敏、小麦依赖运动诱发的过敏反应、特应性皮炎和乳糜泻（即麸质敏感肠病）的关系。总之，结果表明，MS(E)方法适合于小麦品种和/或其他食物基质中的定量分析和过敏原分析。
