In silico identification of IgE-binding epitopes of osmotin protein.
The identification of B-cell epitopes is an important step to study the antigen- antibody interactions for diagnosis and therapy. The present study aimed to identify B- cell epitopes of osmotin using bioinformatic tools and further modify these regions to study the allergenic property. B-cell epitop...
| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2013-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC3548786?pdf=render |
| _version_ | 1811268742735200256 |
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| author | Prerna Sharma Shailendra Nath Gaur Naveen Arora |
| author_facet | Prerna Sharma Shailendra Nath Gaur Naveen Arora |
| author_sort | Prerna Sharma |
| collection | DOAJ |
| description | The identification of B-cell epitopes is an important step to study the antigen- antibody interactions for diagnosis and therapy. The present study aimed to identify B- cell epitopes of osmotin using bioinformatic tools and further modify these regions to study the allergenic property. B-cell epitopes were predicted based on amino acid physicochemical properties. Three single point mutations M1, M2, and M3 and a multiple point mutant (M123) were selected to disrupt the IgE binding. These mutants were cloned, expressed and proteins purified to homogeneity. The IgE binding of the purified proteins was evaluated by ELISA and ELISA inhibition with patients' sera. Three regions of osmotin M1 (57-70 aa), M2 (72-85 aa) and M3 (147-165 aa) were identified as potential antibody recognition sites using in silico tools. The sequence similarity search of the predicted epitopes of osmotin using Structural Database of Allergenic proteins (SDAP) showed similarity with known allergens from tomato, kiwifruit, bell pepper, apple, mountain cedar and cypress. Mutants M1, M2 and M3 showed up to 72%, 60% and 76% reduction, respectively in IgE binding whereas M123 showed up to 90% reduction with patients' sera. The immunoblot of M123 mutant showed 40% reduction in spot density as compared to osmotin. All mutants showed decreased inhibition potency with M123 exhibiting lowest potency of 32% with osmotin positive pooled patients' sera. The three B- cell epitopes of osmotin predicted by in silico method correlated with the experimental approach. The mutant M123 showed a reduction of 90% in IgE binding. The present method may be employed for prediction of B- cell epitopes of allergenic proteins. |
| first_indexed | 2024-04-12T21:28:22Z |
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| id | doaj.art-be615e25d97f4f6d9d552648b4ed8423 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-04-12T21:28:22Z |
| publishDate | 2013-01-01 |
| publisher | Public Library of Science (PLoS) |
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| series | PLoS ONE |
| spelling | doaj.art-be615e25d97f4f6d9d552648b4ed84232022-12-22T03:16:07ZengPublic Library of Science (PLoS)PLoS ONE1932-62032013-01-0181e5475510.1371/journal.pone.0054755In silico identification of IgE-binding epitopes of osmotin protein.Prerna SharmaShailendra Nath GaurNaveen AroraThe identification of B-cell epitopes is an important step to study the antigen- antibody interactions for diagnosis and therapy. The present study aimed to identify B- cell epitopes of osmotin using bioinformatic tools and further modify these regions to study the allergenic property. B-cell epitopes were predicted based on amino acid physicochemical properties. Three single point mutations M1, M2, and M3 and a multiple point mutant (M123) were selected to disrupt the IgE binding. These mutants were cloned, expressed and proteins purified to homogeneity. The IgE binding of the purified proteins was evaluated by ELISA and ELISA inhibition with patients' sera. Three regions of osmotin M1 (57-70 aa), M2 (72-85 aa) and M3 (147-165 aa) were identified as potential antibody recognition sites using in silico tools. The sequence similarity search of the predicted epitopes of osmotin using Structural Database of Allergenic proteins (SDAP) showed similarity with known allergens from tomato, kiwifruit, bell pepper, apple, mountain cedar and cypress. Mutants M1, M2 and M3 showed up to 72%, 60% and 76% reduction, respectively in IgE binding whereas M123 showed up to 90% reduction with patients' sera. The immunoblot of M123 mutant showed 40% reduction in spot density as compared to osmotin. All mutants showed decreased inhibition potency with M123 exhibiting lowest potency of 32% with osmotin positive pooled patients' sera. The three B- cell epitopes of osmotin predicted by in silico method correlated with the experimental approach. The mutant M123 showed a reduction of 90% in IgE binding. The present method may be employed for prediction of B- cell epitopes of allergenic proteins.http://europepmc.org/articles/PMC3548786?pdf=render |
| spellingShingle | Prerna Sharma Shailendra Nath Gaur Naveen Arora In silico identification of IgE-binding epitopes of osmotin protein. PLoS ONE |
| title | In silico identification of IgE-binding epitopes of osmotin protein. |
| title_full | In silico identification of IgE-binding epitopes of osmotin protein. |
| title_fullStr | In silico identification of IgE-binding epitopes of osmotin protein. |
| title_full_unstemmed | In silico identification of IgE-binding epitopes of osmotin protein. |
| title_short | In silico identification of IgE-binding epitopes of osmotin protein. |
| title_sort | in silico identification of ige binding epitopes of osmotin protein |
| url | http://europepmc.org/articles/PMC3548786?pdf=render |
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