title: Synthesis of hypoallergenic derivatives of the major allergen Fag t 1 from tartary buckwheat via sequence restructuring.
abstract: Fag t 1, a legumin-type protein, is the major allergen in tartary buckwheat. In the current study, three recombinant derivatives of Fag t 1, designated as Fag t 1-rs1, Fag t 1-rs2, and Fag t 1-rs3, were constructed via rational design and genetic engineering. However, because of the loss of their native-like folds, the Fag t 1 derivatives failed to bind IgE, and their allergenic activities were reduced. The recombinant hypoallergenic variants are promising vaccine candidates for specific immunotherapy of buckwheat allergy. The unfolding of the Fag t 1 structure reduced its high resistance to gastrointestinal proteolysis and strongly reduced its IgE reactivity. The derivatives showed a more than 90% reduction in allergenic activity compared with rFag t 1. These results suggest that the structure-dependent stability of 11S seed storage proteins is directly related to digestive stability and allergenic potential. Therefore, the destruction of the native conformation is the appropriate strategy to reduce the allergenicity of the cupin family food allergens.
authors.affiliation: Key Laboratory of Chemical Biology and Molecular Engineering of Ministry of Education, Institute of Biotechnology, Shanxi University, Taiyuan 030006, PR China.
authors.forename: Zhenhuang
authors.initials: Z
authors.lastname: Yang
authors.forename: Yuying
authors.initials: Y
authors.lastname: Li
authors.forename: Chen
authors.initials: C
authors.lastname: Li
authors.forename: Zhuanhua
authors.initials: Z
authors.lastname: Wang
mesh_terms: D000485:Allergens / Q000737:chemistry* / Q000276:immunology; D000595:Amino Acid Sequence; D001483:Base Sequence; D015153:Blotting, Western; D002942:Circular Dichroism; D017931:DNA Primers; D004063:Digestion; D004797:Enzyme-Linked Immunosorbent Assay; D019612:Fagopyrum / Q000276:immunology*; D006801:Humans; D008969:Molecular Sequence Data; D013056:Spectrophotometry, Ultraviolet
chemical_list: D000485:Allergens; D017931:DNA Primers