title: Immunological characterization of recombinant soy protein allergen produced by Escherichia coli expression system.
abstract: To elucidate the molecular mechanism of the allergenicity of soybean P34 protein recognized as the most allergenic protein in soybean, the protein was expressed in Escherichia coli transformed with a plasmid carrying P34 cDNA. SDS-PAGE pattern showed that the molecular weight of the recombinant P34 was approximately 2 kDa less than that of the native soybean P34. The difference in the molecular mass between these two proteins could be due to the native P34 in soybean being glycosylated at position Asn(170), whereas the recombinant protein generated in E. coli lacks this post-translational modification. Immunoblot analysis showed that both soybean and recombinant P34 proteins cross-reacted not only with polyclonal and monoclonal antibodies produced against P34 and crude soybean protein but also with patients' sera. The results suggest that the recombinant P34 is immunologically reactive, indicating that both proteins have similar epitope structures. Thus, the recombinant P34 produced by the E. coli expression system can be used as a standard allergen for molecular design to reduce the allergenic structure.
authors.affiliation: Department of Biological Chemistry, Yamaguchi University, Yamaguchi 753, Japan, and Research Institute for Food Science, Kyoto University, Uji, Kyoto 611, Japan.
authors.forename: E E
authors.initials: EE
authors.lastname: Babiker
authors.forename: H
authors.initials: H
authors.lastname: Azakami
authors.forename: T
authors.initials: T
authors.lastname: Ogawa
authors.forename: A
authors.initials: A
authors.lastname: Kato
mesh_terms: D000485:Allergens / Q000276:immunology*; D000818:Animals; D003876:Dermatitis, Atopic / Q000276:immunology; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000378:metabolism*; D006801:Humans; D051379:Mice; D008809:Mice, Inbred C3H; D011817:Rabbits; D011994:Recombinant Proteins / Q000276:immunology; D030262:Soybean Proteins / Q000276:immunology*
chemical_list: D000485:Allergens; D011994:Recombinant Proteins; D030262:Soybean Proteins