Engineering of hypoallergenic mutants of the Brassica pollen allergen, Bra r 1, for immunotherapy

The Brassica pollen allergen Bra r 1 belongs to a new family of Ca²⁺-binding proteins, characterized by the presence of two potential EF-hand calcium-binding domains. Disruption of these EF-hand motifs by amino acid substitutions demonstrated that both domains of Bra r 1 constitute functional Ca²⁺-binding sites. Calcium-binding deficient mutants displayed significantly reduced IgE-binding activity. Injection of these mutated Bra r 1 variants into a murine model system showed that mouse IgG raised against the mutants recognized native Bra r 1 in Brassica pollen extracts suggesting the potential use of the engineered allergens for effective immunotherapy. Copyright (C) 1998 Federation of European Biochemical Societies.