Koppelman S J, Knol E F, Vlooswijk R A A, Wensing M, Knulst A C, Hefle S L, Gruppen H, Piersma S
TNO Nutrition and Food Research Institute, Zeist, The Netherlands.
Allergy. 2003 Nov;58(11):1144-51. doi: 10.1034/j.1398-9995.2003.00259.x.
Peanut allergen Ara h 3 has been the subject of investigation for the last few years. The reported data strongly depend on recombinant Ara h 3, since a purification protocol for Ara h 3 from peanuts was not available.
Peanut allergen Ara h 3 (glycinin), was purified and its posttranslational processing was investigated. Its allergenic properties were determined by studying IgE binding characteristics of the purified protein.
Ara h 3 consists of a series of polypeptides ranging from approximately 14 to 45 kDa that can be classified as acidic and basic subunits, similar to the subunit organization of soy glycinin. N-terminal sequences of the individual polypeptides were determined, and using the cDNA deduced amino-acid sequence, the organization into subunits was explained by revealing posttranslational processing of the different polypeptides. IgE-binding properties of Ara h 3 were investigated using direct elisa and Western blotting with sera from peanut-allergic individuals. The basic subunits, and to a lesser extent the acidic subunits, bind IgE and may act as allergenic peptides.
We conclude that peanut-derived Ara h 3, in contrast to earlier reported recombinant Ara h 3, resembles, to a large extent, the molecular organization typical for proteins from the glycinin family. Furthermore, posttranslational processing of Ara h 3 affects the IgE-binding properties and is therefore an essential subject of study for research on the allergenicity of Ara h 3.
花生过敏原Ara h 3在过去几年一直是研究的对象。由于无法从花生中获得Ara h 3的纯化方案，所报道的数据很大程度上依赖于重组Ara h 3。
纯化花生过敏原Ara h 3（大豆球蛋白），并研究其翻译后加工过程。通过研究纯化蛋白的IgE结合特性来确定其致敏特性。
Ara h 3由一系列分子量约为14至45 kDa的多肽组成，可分为酸性和碱性亚基，类似于大豆球蛋白的亚基组成。测定了各个多肽的N端序列，并利用cDNA推导的氨基酸序列，通过揭示不同多肽的翻译后加工过程解释了亚基的组成。使用直接酶联免疫吸附测定法（ELISA）和来自花生过敏个体血清的蛋白质印迹法研究了Ara h 3的IgE结合特性。碱性亚基以及在较小程度上的酸性亚基结合IgE，并可能作为致敏肽起作用。
我们得出结论，与早期报道的重组Ara h 3相比，花生来源的Ara h 3在很大程度上类似于大豆球蛋白家族蛋白质的典型分子结构。此外，Ara h 3的翻译后加工影响IgE结合特性，因此是研究Ara h 3致敏性的重要研究课题。
