Although Cupressus sempervirens (Cups) pollen represents one of the main aeroallergens in southern Europe, only two Cups allergens have yet been identified and reported: Cup s 1 and Cup s 3.The aim of this study was to identify allergens in cypress pollen using an immuno-proteomic approach. A sequential pollen protein extraction was developed and supplemented by a combinatorial peptide ligand library (CPLL) treatment to select low-abundance species. Control extracts and CPLL eluates have then been resolved by 1-DE and 2-DE gel electrophoresis, blotted and confronted with sera from cypress allergic patients. Extracted proteins including IgE-binding components were identified using nanoLC-MS/MS analysis. A total of 108 unique gene products were identified analyzing the eluates and control loaded onto 1-DE SDS-PAGE. Forty proteins were identified in control samples and 68 supplementary species upon CPLL treatment. Out of the 12 IgE-binding proteins characterized in 2-DE gels, 9 were already reported as allergens in various sources including the two major known allergens of Cupressaceae (groups 1 and 2). Three IgE-binding proteins, not previously reported as allergens, are newly described. The improvement in protein extraction combined with the enrichment of low-abundance species allowed us to extend the repertoire of potential cypress pollen allergens.
Allergomic study of cypress pollen via combinatorial peptide ligand libraries / Y. Shahali, J. Sutra, E. Fasoli, A. D'Amato, P. Righetti, N. Futamura, E. Boschetti, H. Senechal, P. Poncet. - In: JOURNAL OF PROTEOMICS. - ISSN 1874-3919. - 77(2012 Dec 21), pp. 101-110.
|Titolo:||Allergomic study of cypress pollen via combinatorial peptide ligand libraries|
|Parole Chiave:||Cupressus sempervirens; Cypress pollen allergens; Low-abundance proteome; Mass spectrometry; Peptide ligand libraries; Proteomics; Biophysics; Biochemistry|
|Settore Scientifico Disciplinare:||Settore BIO/10 - Biochimica|
Settore CHIM/01 - Chimica Analitica
|Data di pubblicazione:||21-dic-2012|
|Digital Object Identifier (DOI):||http://dx.doi.org/10.1016/j.jprot.2012.07.010|
|Appare nelle tipologie:||01 - Articolo su periodico|