Identification of four IgE-reactive proteins in raspberry (Rubus ideaeus L.).
IgE-reactive proteins in raspberry (Rubus ideaus L.) were identified using PCR, RT-PCR, 2-DE and MS/MS peptide sequencing. Specific polyclonal antibodies and patient sera were used in Western blotting to identify crossreactive epitopes. Initially, two potential allergens Rub i 1 and Rub i 3 were detected using PCR, showing high sequence identity to proteins in Rosaceous species like Mal d 1 and Mal d 3 from apple, Pru av 1 and Pru av 3 from cherry and Pru p 1 and Pru p 3 from peach. Furthermore, de novo identified peptides of a protein band at about 30 kDa reacting with most of the patient sera tested (> 80%) revealed a high sequence homology with class III chitinases. Raspberry chitinase, when subjected to glycoproteomic analysis, showed typical complex plant-type N-glycans with a core alpha1,3 fucose and a beta1,2 xylose at least at one position, indicating the presence of crossreacting carbohydrate determinants (CCDs). Finally, MS/MS analysis revealed an IgE-reactive raspberry cyclophilin, homologous to Bet v 7. Results obtained suggest that the consumption of raspberries might be responsible for adverse reactions in sensitised individuals.
Marzban
Gorji
G
Plant Biotechnology Unit, Department of Biotechnology, BOKU, Vienna, Austria.
Herndl
Anita
A
Kolarich
Daniel
D
Maghuly
Fatemeh
F
Mansfeld
Agata
A
Hemmer
Wolfgang
W
Katinger
Hermann
H
Laimer
Margit
M
D000328:Adult; D000485:Allergens / Q000032:analysis* / Q000737:chemistry; D000595:Amino Acid Sequence; D002688:Chitinases / Q000032:analysis / Q000737:chemistry; D005260:Female; D005512:Food Hypersensitivity / Q000209:etiology; D006801:Humans; D007073:Immunoglobulin E / Q000276:immunology*; D008875:Middle Aged; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000032:analysis*; D027824:Rosaceae / Q000276:immunology*; D016415:Sequence Alignment
D000485:Allergens; D010940:Plant Proteins; D007073:Immunoglobulin E; D002688:Chitinases; C110892:chitinase, class III