Rapid isolation, characterization, and glycan analysis of Cup a 1, the major all_ergen of Arizona cypress (Cupressus arizonica) pollen

• DOI: 10.1034/j.1398-9995.2001.103125.x
• Publication Date: 2003-06-06T14:03:39Z
• Authors: C. Alisi; C. Afferni; P. Iacovacci; B. Barletta; R. Tinghino; C. Butteroni; E. M. R. Puggioni; I. B. H. Wilson; R. Federico; M. E. Schininà; R. Ariano; G. Di Felice; C. Pini
• Keywords: Cypress; Oligosaccharide

A rapid method for the purification of major 43-kDa allergen Cupressus arizonica pollen, Cup a 1, was developed.The salient feature wash pollen in acidic buffer, followed by an extraction proteins and their chromatography. Immunoblotting, ELISA, lectin binding were tested on both crude extract purified 1. Biochemical analyses performed to assess 1 isoelectric point, its partial amino-acid sequence, glycan composition.Immunochemical analysis confirmed that allergenic reactivity is maintained after process. Partial sequencing indicated high degree homology between from Cupressaceae Taxodiaceae families displaying similar molecular mass. The protein shows one band with point 5.2. Nineteen out 33 sera (57%) patients allergic cypress demonstrated significant MALDI-TOF mass spectrometry presence three N-linked oligosaccharide structures: GnGnXF(3) (i.e., horseradish peroxidase-type substituted two nonreducing N-acetylglucosamine residues), GGnXF(3)/GnGXF(3) GnGnXF galactose residue), (GF)GnXF(3)/Gn(GF)XF(3) (with Lewisa epitope arm) molar ratio 67:8:23.The process allowed some fine studies properties structure, as well evaluation IgE native conditions. similarities sequences complex stuctures could explain cross-reactivity among families.