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Immunoproteomic and two-dimensional difference gel electrophoresis analysis of Arabidopsis dehydration response element-binding protein 1A (DREB1A)-transgenic potato.
- Source :
-
Biological & pharmaceutical bulletin [Biol Pharm Bull] 2010; Vol. 33 (8), pp. 1418-25. - Publication Year :
- 2010
-
Abstract
- To produce crops that are more tolerant to stresses such as heat, cold, and salt, transgenic plants have been produced those express stress-associated proteins. In this study, we used immunoproteomic and two-dimensional difference gel electrophoresis (2D-DIGE) methods to investigate the allergenicity of transgenic potatoes expressing Arabidopsis DREB1A (dehydration responsive element-binding protein 1A), driven by the rd29A promoter or the 35S promoter. Immunoproteomic analysis using sera from potato-allergic patients revealed several immunoglobulin E (IgE)-binding protein spots. The patterns of protein binding were almost the same between transgenic and non-transgenic potatoes. The IgE-binding proteins in potato were identified as patatin precursors, a segment of serine protease inhibitor 2, and proteinase inhibitor II by matrix assisted laser desorption/ionization-time of flight (MALDI-TOF) MS/MS. 2D-DIGE analysis revealed several differences in protein expression between non-transgenic potato and transgenic potato; those showing increased expression in transgenic potatoes were identified as precursors of patatin, a major potato allergen, and those showing decreased expression in transgenic potatoes were identified as lipoxygenase and glycogen (starch) synthase. These results suggested that transgenic potatoes may express slightly higher levels of allergens, but their IgE-binding patterns were almost the same as those of control potatoes. Further research on changes in protein expressions in response to environmental factors is required to confirm whether the differences observed in this study are due to gene transfection, rather than environmental factors.
- Subjects :
- Allergens genetics
Allergens immunology
Carboxylic Ester Hydrolases genetics
Carboxylic Ester Hydrolases immunology
Electrophoresis, Gel, Two-Dimensional
Electrophoresis, Polyacrylamide Gel
Food Hypersensitivity blood
Humans
Immunoblotting
Plant Proteins genetics
Plant Proteins immunology
Protein Binding
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
Transfection
Transgenes
Arabidopsis Proteins genetics
Food Hypersensitivity immunology
Immunoglobulin E blood
Plants, Genetically Modified adverse effects
Plants, Genetically Modified genetics
Plants, Genetically Modified immunology
Solanum tuberosum adverse effects
Solanum tuberosum genetics
Solanum tuberosum immunology
Transcription Factors genetics
Subjects
Details
- Language :
- English
- ISSN :
- 1347-5215
- Volume :
- 33
- Issue :
- 8
- Database :
- MEDLINE
- Journal :
- Biological & pharmaceutical bulletin
- Publication Type :
- Academic Journal
- Accession number :
- 20686241
- Full Text :
- https://doi.org/10.1248/bpb.33.1418