title: IgE binding studies with large peptides expressed from Der p II cDNA constructs.
abstract: The major mite allergen Der p II shows marked resistance to denaturation and is expressed from cDNA in bacteria with almost all of its IgE binding activity. Despite these properties, the IgE binding activity appears to be dependent on maintaining the complete primary structure. Random fragment libraries of cDNA, able to code for up to 93 of the 129 amino acid residue protein, did not express IgE binding peptides. Large overlapping peptides 1-69, 69-129 and 42-117 expressed as the fusions from the glutathione transferase of pGEX vectors only had binding activity with IgE in 15 out of 57 sera, and this was typically weak. Sera from children with atopic dermatitis bound IgE in seven out of eight cases but this was also weak compared with their strong reactivity to intact recombinant Der p II. The inability of such large peptides to form IgE binding structures suggests that the antigenic determinants of Der p II are highly conformational and restricted.
authors.affiliation: Western Australian Research Institute for Child Health, Princess Margaret Hospital for Children, Subiaco.
authors.forename: K Y
authors.initials: KY
authors.lastname: Chua
authors.forename: W K
authors.initials: WK
authors.lastname: Greene
authors.forename: P
authors.initials: P
authors.lastname: Kehal
authors.forename: W R
authors.initials: WR
authors.lastname: Thomas
mesh_terms: D000485:Allergens / Q000235:genetics*; D000818:Animals; D039741:Antigens, Dermatophagoides; D004247:DNA / Q000235:genetics; D000939:Epitopes / Q000235:genetics; D006801:Humans; D006967:Hypersensitivity / Q000276:immunology; D007073:Immunoglobulin E / Q000378:metabolism; D008925:Mites / Q000235:genetics / Q000276:immunology*; D015183:Restriction Mapping
chemical_list: D000485:Allergens; D039741:Antigens, Dermatophagoides; D000939:Epitopes; D007073:Immunoglobulin E; D004247:DNA