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.
Okada
T
T
Laboratory of Plant Breeding and Genetics, Graduate School of Agricultural Science, Tohoku University, Sendai, Japan.
Swoboda
I
I
Bhalla
P L
PL
Toriyama
K
K
Singh
M B
MB
D000485:Allergens / Q000235:genetics* / Q000378:metabolism / Q000627:therapeutic use; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001937:Brassica / Q000276:immunology*; D002118:Calcium / Q000378:metabolism; D017931:DNA Primers; D004797:Enzyme-Linked Immunosorbent Assay; D007073:Immunoglobulin E / Q000096:biosynthesis; D007167:Immunotherapy; D051379:Mice; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D011058:Pollen / Q000276:immunology*; D011485:Protein Binding; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism / Q000627:therapeutic use
D000485:Allergens; D017931:DNA Primers; D011994:Recombinant Proteins; D007073:Immunoglobulin E; D002118:Calcium