Identification of the amino‐terminal fragment of Ara h 1 as a major target of the IgE‐binding activity in the basic peanut protein fraction

Background Small, basic peanut proteins are often poorly extracted in pH‐neutral buffers that are optimal for the extraction of peanut storage proteins such as Ara h 1. As a result, such proteins are easily missed as potential allergens. Objective To analyse the allergenic composition of the basic p...

Full description

Saved in:
Bibliographic Details
Published inClinical and experimental allergy Vol. 50; no. 3; pp. 401 - 405
Main Authors Aalberse, Rob C., Mueller, Geoffrey A., Derksen, Ninotska I. L., Aalberse, Joost A., Edwards, Lori L., Pomés, Anna, Lidholm, Jonas, Rispens, Theo, Briza, Peter
Format Journal Article
LanguageEnglish
Published England Wiley Subscription Services, Inc 01.03.2020
Subjects
Allergens
allergens and epitopes
Ara h 1 antigen
Food allergies
food allergy
Gel electrophoresis
IgE
Immunoblotting
Immunoglobulin E
immunologic tests
Mass spectroscopy
pH effects
Proteins
Sodium lauryl sulfate
Storage proteins
immunologic tests
allergens and epitopes
food allergy
IgE
Online AccessGet full text

Cover

More Information
Summary:Background Small, basic peanut proteins are often poorly extracted in pH‐neutral buffers that are optimal for the extraction of peanut storage proteins such as Ara h 1. As a result, such proteins are easily missed as potential allergens. Objective To analyse the allergenic composition of the basic peanut protein (BPP) fraction. Methods A peanut extract prepared at pH 4 was fractionated by physicochemical procedures. Chemical analysis was performed by SDS‐PAGE and mass spectrometry. Because immunoblotting was found to be inefficient for most of these small basic proteins, IgE‐binding activity was measured by coupling the fractions to CNBr‐activated Sepharose, followed by incubation with sera from 55 Dutch peanut‐allergic children and 125I‐labelled anti‐IgE. Results Most IgE reactivity of the BPP fraction was due to the 5‐7 kDa amino‐terminal fragment of Ara h 1. This finding was confirmed by the use of the fragment in recombinant form, to which 25/55 of the sera was IgE‐positive. Conclusion The amino‐terminal fragment of Ara h 1, a member of a family of small anti‐microbial proteins, is an allergen independent of the carboxy‐terminal fragment of Ara h 1.
Bibliography:Funding information
This study was supported in part by the Intramural Research Program of the National Institute of Environmental Health Sciences (Z01‐ES102906‐01, G Mueller).
RA, PB, TR, GM, AP and JL contributed to the underlying concept and study design. ND, LE, PB, TR, GM and RA performed the experiments and data analyses. JA was responsible for the clinical work. RA wrote the initial draft of the manuscript. All coauthors approved the final version of the manuscript.
Author contributions
ISSN:0954-7894
1365-2222
DOI:10.1111/cea.13554