Title
Peanut allergen Ara h 3: Isolation from peanuts and biochemical characterization
Author
Koppelman, S.J.
Knol, E.F.
Vlooswijk, R.A.A.
Wensing, M.
Knulst, A.C.
Hefle, S.L.
Gruppen, H.
Piersma, S.
Publication year
2003
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.
Subject
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
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DOI
https://doi.org/10.1034/j.1398-9995.2003.00259.x
TNO identifier
237356
ISSN
0105-4538
Source
Allergy: European Journal of Allergy and Clinical Immunology, 58 (11), 1144-1151
Document type
article