title: IgE binding conformational epitopes of Asp f 3, a major allergen of Aspergillus fumigatus.
abstract: sp f 3 has been identified as one of the major allergens of Aspergillus fumigatus associated with the sensitization and immune responses in allergic bronchopulmonary aspergillosis (ABPA). In order to understand the structure/function relationship of Asp f 3, we studied synthetic peptides and constructed mutants deleted of specific IgE binding regions. The mutated allergens were obtained by expressing the genes and studied by ELISA for their reactivity with IgE from patients with ABPA. Seven linear IgE binding regions spanning the whole Asp f 3 molecule were demonstrated. The results demonstrated strong binding of IgE from ABPA patients with Asp f 3 and one mutant, Asp f 3(1-150), but not with other mutant constructs. The results identified 12 amino acids at the N-terminal end and 8 amino acids (143-150) at the C-terminal end as significant in the conformational constraints for IgE binding. The Fourier transfer spectra showed comparable beta-sheet structure of Asp f 3(1-150) and Asp f 3, indicating the role of secondary structure in IgE binding. The primary and secondary structures may help understanding of the functional role the allergens play in the disease and may have implications in immunodiagnosis and probably immunotherapy.
authors.affiliation: Department of Pediatrics, Allergy-Immunology Division, Medical College of Wisconsin, Milwaukee, Wisconsin, USA.
authors.forename: Harikrishnan
authors.initials: H
authors.lastname: Ramachandran
authors.forename: Vasanthi
authors.initials: V
authors.lastname: Jayaraman
authors.forename: Banani
authors.initials: B
authors.lastname: Banerjee
authors.forename: Paul A
authors.initials: PA
authors.lastname: Greenberger
authors.forename: Kevin J
authors.initials: KJ
authors.lastname: Kelly
authors.forename: Jordan N
authors.initials: JN
authors.lastname: Fink
authors.forename: Viswanath P
authors.initials: VP
authors.lastname: Kurup
mesh_terms: D000485:Allergens / Q000235:genetics / Q000276:immunology*; D000595:Amino Acid Sequence; D052179:Antigens, Plant; D001232:Aspergillus fumigatus / Q000235:genetics / Q000276:immunology*; D015153:Blotting, Western; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000276:immunology; D004926:Escherichia coli / Q000235:genetics; D006801:Humans; D007073:Immunoglobulin E / Q000097:blood / Q000276:immunology* / Q000378:metabolism; D008969:Molecular Sequence Data; D009154:Mutation; D017433:Protein Structure, Secondary; D011994:Recombinant Proteins; D017550:Spectroscopy, Fourier Transform Infrared
chemical_list: D000485:Allergens; D052179:Antigens, Plant; C115315:Asp f 3 allergen; D000939:Epitopes; D011994:Recombinant Proteins; D007073:Immunoglobulin E