Phenoxy radical reactivity of nucleic acids: practical implications for biotinylation
Recent advances in peroxidase-mediated biotin tyramide (BT) signal amplification technology have resulted in high-resolution and subcellular compartment-specific mapping of protein and RNA localization. Horseradish peroxidase (HRP) in the presence of H O is known to activate phenolic compounds for...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Wiley
2021
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| Online Access: | https://hdl.handle.net/1721.1/133515 |
| _version_ | 1826200100044537856 |
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| author | Wilbanks, Brandon Garcia, Brian Byrne, Shane Dedon, Peter Maher, L James |
| author_facet | Wilbanks, Brandon Garcia, Brian Byrne, Shane Dedon, Peter Maher, L James |
| author_sort | Wilbanks, Brandon |
| collection | MIT |
| description | Recent advances in peroxidase-mediated biotin tyramide (BT) signal amplification technology have resulted in high-resolution and subcellular compartment-specific mapping of protein and RNA localization. Horseradish peroxidase (HRP) in the presence of H O is known to activate phenolic compounds for phenoxy radical reaction with nucleic acids, where biotinylation by BT is a practical example. BT reactivity with RNA and DNA is not understood in detail. We report that BT phenoxy radicals react in a sequence-independent manner with guanosine bases in RNA. In contrast, DNA reactivity with BT cannot be detected by our methods under the same conditions. Remarkably, we show that fluorescein conjugates DNA rapidly and selectively reacts with BT phenoxy radicals, allowing convenient and practical biotinylation of DNA on fluorescein with retention of fluorescence. 2 2 |
| first_indexed | 2024-09-23T11:31:11Z |
| format | Article |
| id | mit-1721.1/133515 |
| institution | Massachusetts Institute of Technology |
| language | English |
| last_indexed | 2024-09-23T11:31:11Z |
| publishDate | 2021 |
| publisher | Wiley |
| record_format | dspace |
| spelling | mit-1721.1/1335152021-10-28T04:15:29Z Phenoxy radical reactivity of nucleic acids: practical implications for biotinylation Wilbanks, Brandon Garcia, Brian Byrne, Shane Dedon, Peter Maher, L James Recent advances in peroxidase-mediated biotin tyramide (BT) signal amplification technology have resulted in high-resolution and subcellular compartment-specific mapping of protein and RNA localization. Horseradish peroxidase (HRP) in the presence of H O is known to activate phenolic compounds for phenoxy radical reaction with nucleic acids, where biotinylation by BT is a practical example. BT reactivity with RNA and DNA is not understood in detail. We report that BT phenoxy radicals react in a sequence-independent manner with guanosine bases in RNA. In contrast, DNA reactivity with BT cannot be detected by our methods under the same conditions. Remarkably, we show that fluorescein conjugates DNA rapidly and selectively reacts with BT phenoxy radicals, allowing convenient and practical biotinylation of DNA on fluorescein with retention of fluorescence. 2 2 2021-10-27T19:53:17Z 2021-10-27T19:53:17Z 2021 2021-08-26T16:32:36Z Article http://purl.org/eprint/type/JournalArticle https://hdl.handle.net/1721.1/133515 en 10.1002/CBIC.202000854 ChemBioChem Creative Commons Attribution-NonCommercial-NoDerivs License http://creativecommons.org/licenses/by-nc-nd/4.0/ application/pdf Wiley Wiley |
| spellingShingle | Wilbanks, Brandon Garcia, Brian Byrne, Shane Dedon, Peter Maher, L James Phenoxy radical reactivity of nucleic acids: practical implications for biotinylation |
| title | Phenoxy radical reactivity of nucleic acids: practical implications for biotinylation |
| title_full | Phenoxy radical reactivity of nucleic acids: practical implications for biotinylation |
| title_fullStr | Phenoxy radical reactivity of nucleic acids: practical implications for biotinylation |
| title_full_unstemmed | Phenoxy radical reactivity of nucleic acids: practical implications for biotinylation |
| title_short | Phenoxy radical reactivity of nucleic acids: practical implications for biotinylation |
| title_sort | phenoxy radical reactivity of nucleic acids practical implications for biotinylation |
| url | https://hdl.handle.net/1721.1/133515 |
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