title: Proteomic profiling of birch (Betula verrucosa) pollen extracts from different origins.
abstract: Pollen of the European white birch is a major source of spring pollinosis in Europe. Pollen-allergy diagnosis and treatment by specific immunotherapy commonly rely on extracts of natural origin. To gain insight into the protein content and its variability, we evaluated the profile of allergenic and non-allergenic proteins in extracts of pollen from different origins by MS-based proteomics. Aqueous extracts prepared from commercially available Swedish birch pollen, pollen collected from Austrian trees and a commercial skin prick extract were analyzed by 1-DE, 2-DE, immunoblotting and mass spectrometry, resulting in a complete inventory of extractable, disease-relevant pollen proteins. A main focus of this study was on the isoform distribution of Bet v 1, the major allergen of birch pollen. Using a combination of intact mass determination and peptide sequencing, five isoforms (a, b, d, f and j) were unequivocally identified in Swedish and Austrian birch pollen extracts, while the skin prick extract contained only isoforms a, b and d. Using the same methods as for Bet v 1, divergencies in the sequence of birch profilin (Bet v 2), a plant panallergen, were solved. The molecular characterization of pollen extracts is relevant for standardization and development of new reagents for specific immunotherapy.
authors.affiliation: Christian Doppler Laboratory for Allergy Diagnosis and Therapy, Department of Molecular Biology, University of Salzburg, Salzburg, Austria.
authors.forename: Anja
authors.initials: A
authors.lastname: Erler
authors.forename: Thomas
authors.initials: T
authors.lastname: Hawranek
authors.forename: Leif
authors.initials: L
authors.lastname: Krückemeier
authors.forename: Claudia
authors.initials: C
authors.lastname: Asam
authors.forename: Matthias
authors.initials: M
authors.lastname: Egger
authors.forename: Fátima
authors.initials: F
authors.lastname: Ferreira
authors.forename: Peter
authors.initials: P
authors.lastname: Briza
mesh_terms: D000485:Allergens / Q000737:chemistry*; D052179:Antigens, Plant / Q000032:analysis; D029662:Betula / Q000737:chemistry*; D015151:Immunoblotting; D013058:Mass Spectrometry; D010936:Plant Extracts / Q000032:analysis*; D010940:Plant Proteins / Q000032:analysis; D011058:Pollen / Q000737:chemistry*; D020033:Protein Isoforms; D040901:Proteomics
chemical_list: D000485:Allergens; D052179:Antigens, Plant; D010936:Plant Extracts; D010940:Plant Proteins; D020033:Protein Isoforms