Proteomic analysis of known and candidate rice allergens between non-transgenic and transgenic plants

• Published in: Regulatory toxicology and pharmacology Vol. 59; no. 3; pp. 437 - 444
• Main Authors: Satoh, Rie, Nakamura, Rika, Komatsu, Akira, Oshima, Masahiro, Teshima, Reiko
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier Inc 01.04.2011
• Subjects: Allergen; Allergens - genetics; Allergens - immunology; Allergens - metabolism; Animals; Binding Sites, Antibody; Humans; IgE-binding protein; Immunoglobulin E - metabolism; Non-transgenic; Oryza - genetics; Oryza - immunology; Oryza sativa; Plant Proteins - genetics; Plant Proteins - immunology; Plant Proteins - metabolism; Plants, Genetically Modified - genetics; Plants, Genetically Modified - immunology; Proteomics; Proteomics - methods; Rice; Seeds - genetics; Seeds - immunology; Transgenic; PBS; MALDI-TOF MS/MS; Non-transgenic; IgE; HRP; Transgenic; SDS–PAGE; PBS-T; KSCN; 2D; 1D; Proteomics; IgE-binding protein; Rice; Allergen
• ISSN: 0273-2300; 1096-0295
• DOI: 10.1016/j.yrtph.2011.01.008

Salt-soluble proteins extracted from non-transgenic and transgenic rice were evaluated for the presence of known and potential allergens by proteomic techniques. The salt-soluble proteins were extracted, separated by 1D and 2D electrophoresis, and analyzed by Western blotting. 1D immunoblot analysis with patients’ sera revealed few qualitative differences between the IgE-binding proteins of the non-transgenic and transgenic rice. 1D immunoblot with antigen-specific-animal sera revealed no qualitative or quantitative differences in two known allergens, RAG2 and glyoxalase I, between non-transgenic and transgenic rice. Multiple spots containing known and novel IgE-binding proteins were detected among the salt-soluble proteins of non-transgenic rice by 2D immunoblotting. Two globulin-like proteins, a 52kDa protein and a 63kDa protein, were identified as novel IgE-binding proteins that are candidates for rice allergens. These globulin-like proteins were homologous to Cupin superfamily allergens. Quantitative analysis of 19, 52, and 63kDa globulins with protein-specific-animal sera showed no significant differences in the expression of these proteins between the transgenic rice and non-transgenic rice. These results indicate that none of the known or novel endogeneous IgE-binding proteins detected in this study appear to be altered by genetic modification.