Reduction of antigenicity and allergenicity of genetically modified egg white allergen, ovomucoid third domain.
Ovomucoid (Gal d1) is a major allergen in hen egg white, consisting of three tandem domains. In this study, five genetically modified third domain (DIII) mutants, which were substituted single or double amino acids within its IgE and IgG epitopes were compared with those prepared and their antigenicity and allergenicity with native analogue using Western immunoblot and enzyme-linked immunosorbent assay. The replacement of phenylalanine at 37 (F37) position with methionine caused drastical loss of IgG and IgE binding activities of human sera derived from egg allergic patients as well as disruption of the alpha-helix structure which comprises a part of the IgG and IgE epitopes. Substituting glycine at 32 position in conjunction with F37 showed a synergistic effect of decreasing antigenicity. The present study indicated that glycine 32 and phenylalanine 37 have an important role on its antigenicity and allergenicity as well as structural integrity of ovomucoid DIII.
Mine
Yoshinori
Y
Department of Food Science, University of Guelph, Ont., Guelph, Canada N1G2W1. ymine@uoguelph.ca
Sasaki
Erika
E
Zhang
Jie Wei
JW
D000485:Allergens / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D000818:Animals; D000941:Antigens / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D001483:Base Sequence; D015153:Blotting, Western; D002645:Chickens; D017931:DNA Primers; D004797:Enzyme-Linked Immunosorbent Assay; D016297:Mutagenesis, Site-Directed; D010059:Ovomucin / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D017433:Protein Structure, Secondary; D013056:Spectrophotometry, Ultraviolet
D000485:Allergens; D000941:Antigens; D017931:DNA Primers; D010059:Ovomucin