Thermodynamic characterization of the PR-10 allergens Bet v 1, Api g 1 and Dau c 1 and pH-dependence of nApi g 1 and nDau c 1

Natural and recombinant Bet v 1, the major birch pollen allergen, and homologous allergens, Api g 1 and Dau c 1, from celery and carrot, respectively, were studied by CD spectroscopy under conditions of varying denaturant concentration, pH and temperature to determine fundamental thermodynamic parameters for conformational stability. Thermodynamic studies increase basic knowledge regarding differences between birch pollen-related allergens and are of importance in choosing processing conditions. The conformational stability determined from guanidine hydrochloride denaturation curves was similar for rBet v 1.0101 and rApi g 1.0101. Conformational responses to chaotropic salt were different for recombinant allergens from different species, but were similar for the natural isoform mixtures. The conformational stabilities of nApi g 1 and nDau c 1, were shown to be similar to rBet v 1.2801 at pH > 4.4 [Mogensen, J. E., Ipsen, H., Holm, J., & Otzen, D. E. (2004). Elimination of a misfolded folding intermediate by a single point mutation. Biochemistry, 43(12), 3357-3367], but nApi g and nDau c 1 were stable to heating at lower pH-values.