Elucidation of IgE-binding epitopes of Ani s 1: the major Anisakis simplex allergen.
Ani s 1, the major allergen of Anisakis simplex, is expected to be a useful antigen in allergen-specific immunotherapy of Anisakis allergy. To avoid side-effects such as anaphylactic shock in immunotherapy, it is desirable to use not native allergens but hypoallergenic mutants devoid of IgE-binding epitopes. This study was hence aimed to elucidate IgE-binding epitopes of Ani s 1 toward the development of hypoallergenic Ani s 1 mutants. Mapping experiments using 32 peptides (P1-32) revealed that major IgE-binding epitopes are included in P24 (region 116-130) and P28 (region 136-150). Furthermore, Ala-scanning and truncation experiments established that eight (underlined in the sequence 116-ELFAREYEGVCKSGK-130) and nine amino acid residues (underlined in the sequence 136-RGSGWMMTILGKSCD-150) are crucial for the IgE-binding of P24 and P28, respectively. The determined crucial residues of P24 and P28 were assumed to be involved in electrostatic and hydrophobic interactions with IgE, respectively.
Kobayashi
Yukihiro
Y
Department of Food Science and Technology, Tokyo University of Marine Science and Technology, Tokyo, Japan.
Ikeda
Kaori
K
Shiomi
Kazuo
K
D000485:Allergens / Q000737:chemistry / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D017129:Anisakiasis / Q000175:diagnosis / Q000276:immunology; D017163:Anisakis / Q000276:immunology*; D002135:Calcium-Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D004797:Enzyme-Linked Immunosorbent Assay; D018604:Epitope Mapping; D015801:Helminth Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D006801:Humans; D006969:Hypersensitivity, Immediate / Q000175:diagnosis / Q000276:immunology; D007073:Immunoglobulin E / Q000378:metabolism*; D008969:Molecular Sequence Data; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D014581:Urticaria / Q000175:diagnosis / Q000276:immunology
D000485:Allergens; D002135:Calcium-Binding Proteins; D015801:Helminth Proteins; D010455:Peptides; D011994:Recombinant Proteins; C412511:troponin-like protein, Anisakis simplex; D007073:Immunoglobulin E