Attachment of Nitroxide Spin Labels to Nucleic Acids for EPR
| Autor: | Olga Romainczyk, Joachim W. Engels, Xavier Elduque |
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| Rok vydání: | 2012 |
| Předmět: |
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Molecular Nitroxide mediated radical polymerization Pyrimidine Oligonucleotide Chemistry Electron Spin Resonance Spectroscopy Sonogashira coupling DNA General Medicine Site-directed spin labeling Combinatorial chemistry law.invention Cyclic N-Oxides chemistry.chemical_compound Organophosphorus Compounds law Nucleic Acids Fluorescence Resonance Energy Transfer Nucleic acid Nucleic Acid Conformation RNA Spin Labels Electron paramagnetic resonance Solid-Phase Synthesis Techniques Macromolecule |
| Zdroj: | Current Protocols in Nucleic Acid Chemistry |
| ISSN: | 1934-9289 1934-9270 |
| DOI: | 10.1002/0471142700.nc0717s49 |
| Popis: | In addition to X-ray, NMR, and FRET, electron paramagnetic resonance (EPR) can be applied to elucidate the structure of different macromolecular systems and determine local surroundings of paramagnetic centers in DNA and RNA. This technique permits structural characterization as well as dynamic structural changes of macromolecular systems. To do so, free radicals with good stability must be introduced. Here, the site-directed spin labeling of DNA and RNA based on the Sonogashira cross-coupling reaction is described. First, the appropriate building blocks, either 5-iodo-substituted pyrimidine deoxy- or ribo-nucleoside phosphoramidites are prepared and incorporated by solid-phase synthesis. Following this, the protected oligonucleotides are “on column” reacted with the acetylenic nitroxide spin labels and subsequently purified. Applications of this technique for duplexes, hairpins, and riboswitches in vitro and in cell are described. Curr. Protoc. Nucleic Acid Chem. 49:7.17.1-7.17.40. © 2012 by John Wiley & Sons, Inc. Keywords: site-directed spin labeling (SDSL); nitroxide; on-column synthesis; Sonogashira cross coupling |
| Databáze: | OpenAIRE |
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