Structural characterization of the 60-kDa Bermuda grass pollen isoallergens, a covalent flavoprotein

Our studies suggest a tripartite structure for the 60-kDa allergen of Bermu da grass pollen (BG60) including a short N-terminal segment, a FAD binding domain, and a C-terminal domain. The lower molecular weight isoallergens la ck the N-terminal segment. The higher protease susceptibility and the lower melting temperature of approximately 20 degreesC of the lower molecular we ight isoforms suggest that the N-terminal segment is essential for a compac t structure. Database screening reveals that the protease-digested peptide sequences (similar to 180 residues in total) share 40% identity with the pl ant berberine bridge enzymes. In particular, a 24-residue peptide sequence displays high similarity to a conserved FAD-binding motif. The spectroscopi c and SDS-PAGE analyses suggest that the cofactor FAD is covalently linked to the central domain. Therefore, we conclude that BG60 is identified as th e first flavinylated allergen. (C) 2001 Academic Press.