Marsh Justin, Rigby Neil, Wellner Klaus, Reese Gerald, Knulst André, Akkerdaas Jaap, van Ree Ronald, Radauer Christian, Lovegrove Alison, Sancho Ana, Mills Clare, Vieths Stefan, Hoffmann-Sommergruber Karin, Shewry Peter R
Rothamsted Research, Harpenden, Hertfordshire, UK.
Mol Nutr Food Res. 2008 Nov;52 Suppl 2:S272-85. doi: 10.1002/mnfr.200700524.
Peanut is a major cause of type 1 hypersensitive reactions including anaphylaxis. This results from the presence of a number of protein allergens, six of which are being studied as part of the EU FP6 EuroPrevall programme. These are Ara h 1 (7S globulin), Ara h 2, Ara h 6 (2S albumins), Ara h 3/4 (11S globulins) and Ara h 8 (Bet v 1 homologue). Methods for the purification of Ara h 1, Ara h 3/4, Ara h 2 and Ara h 6 from peanut seeds and for the production of recombinant Ara h 8 in Escherichia coli are described with spectroscopic analyses being used to confirm that they are authentically folded. N-terminal sequencing of the proteins purified from peanut seeds also revealed details of the differences between isoforms and their generation by proteolytic processing within the seed. Preliminary IgE binding studies of the purified allergens confirmed that they retained their immunological properties indicating their suitability for use in allergy diagnosis.
花生是包括过敏反应在内的1型超敏反应的主要诱因。这是由于存在多种蛋白质过敏原，其中六种正作为欧盟第六框架计划EuroPrevall项目的一部分进行研究。它们是Ara h 1（7S球蛋白）、Ara h 2、Ara h 6（2S白蛋白）、Ara h 3/4（11S球蛋白）和Ara h 8（Bet v 1同源物）。本文描述了从花生种子中纯化Ara h 1、Ara h 3/4、Ara h 2和Ara h 6的方法，以及在大肠杆菌中生产重组Ara h 8的方法，并通过光谱分析来确认它们的正确折叠。对从花生种子中纯化的蛋白质进行N端测序，也揭示了同工型之间的差异细节以及它们在种子内通过蛋白水解加工产生的过程。对纯化过敏原的初步IgE结合研究证实，它们保留了免疫特性，表明其适用于过敏诊断。
