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Identification and characterisation of allergens related to Bet v I, the major birch pollen allergen, in apple, cherry, celery and carrot by two‐dimensional immunoblotting and N ‐terminal microsequencing
Author(s) -
Schöning Britta,
Vieths Stefan,
Petersen Arnd,
Baltes Werner
Publication year - 1995
Publication title -
journal of the science of food and agriculture
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.782
H-Index - 142
eISSN - 1097-0010
pISSN - 0022-5142
DOI - 10.1002/jsfa.2740670403
Subject(s) - allergen , polyclonal antibodies , chemistry , pollen , polyacrylamide gel electrophoresis , horticulture , botany , biology , biochemistry , antibody , allergy , immunology , enzyme
Low temperature extracts taken from Granny Smith and Braeburn apples, cherries, celery and carrots were separated by two‐dimensional electrophoresis. Bet v I, the major birch pollen allergen, related allergens in apple, cherry, celery and carrot extracts were detected by means of two‐dimensional immunoblotting with patients' sera containing IgE antibodies specific to Bet v I, a rabbit polyclonal antiserum raised against Bet v I , and two Bet v I specific mouse monoclonal antibodies. The major cross‐reacting allergen spots were observed with molecular weights/isoelectric points of 18.0 kDa/p I 5.5 for apple (Granny Smith and Braeburn) and 18.0 kDa/p I 5.8 for cherry, 15.5 kDa/p I 4.4–4.6 for celery and 16.0 kDa/p I 4.4 for carrot extract. Additional antibody reactivities with certain isoprotein spots were observed, which may indicate the presence of Bet v I‐related epitopes on these proteins. Based on the first 15 N‐terminal amino acid residues, the major allergen spots revealed 53% sequence identity between Bet v I and the Granny Smith apple allergen, 50% between Braeburn apple allergen and Bet v I , 67% between Bet v I and the cherry allergen, and 40 and 28% for celery and carrot, respectively. Furthermore, the N‐terminal sequences showed identities ranging from 40% (apple/cherry) to 66% (celery) with PcPR l‐l , a pathogenesis‐related protein in parsley.