Repository hosted by TU Delft Library

Home · Contact · About · Disclaimer ·
 

Peanut allergen Ara h 3: Isolation from peanuts and biochemical characterization

Publication files not online:

Author: Koppelman, S.J. · Knol, E.F. · Vlooswijk, R.A.A. · Wensing, M. · Knulst, A.C. · Hefle, S.L. · Gruppen, H. · Piersma, S.
Type:article
Date:2003
Source:Allergy: European Journal of Allergy and Clinical Immunology, 11, 58, 1144-1151
Identifier: 237356
doi: doi:10.1034/j.1398-9995.2003.00259.x
Keywords: Nutrition Health · Food technology · Allergens · IgE-binding · Peanut · Purification · allergen · Ara h 3 antigen · complementary DNA · glycinin · immunoglobulin E · peanut antigen · protein subunit · unclassified drug · allergenicity · amino acid sequence · amino terminal sequence · antigen binding · article · biochemistry · enzyme linked immunosorbent assay · gel permeation chromatography · nonhuman · peanut · peanut allergy · priority journal · protein analysis · protein folding · protein isolation · protein processing · protein purification · protein structure · protein unfolding · Western blotting · Allergens · Amino Acid Sequence · Arachis hypogaea · Calorimetry, Differential Scanning · Chromatography, Ion Exchange · Electrophoresis, Polyacrylamide Gel · Enzyme-Linked Immunosorbent Assay · Humans · Immunoblotting · Immunoglobulin E · Peanut Hypersensitivity

Abstract

Background: 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. Methods: 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. Results: 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. Conclusions: 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.