| Site-specific incorporation of nitroxide spin-labels into 2'-positions of nucleic acids. | |
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MedLine Citation:
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PMID: 17703207 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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A protocol is described for the incorporation of nitroxide spin-labels into specific 2'-sites within nucleic acids. This labeling strategy facilitates the investigation of nucleic acid structure and dynamics using electron paramagnetic resonance (EPR) spectroscopy and macromolecular complex formation using paramagnetic relaxation enhancement NMR spectroscopy. A spin-labeling reagent, 4-isocyanato TEMPO, which can be prepared in one facile step or obtained commercially, is used for postsynthetic modification of site-specifically 2'-amino-modified nucleic acids. This spin-labeling protocol has been applied primarily to RNA, but is also applicable to DNA. Subsequently, EPR spectroscopic analysis of the spin-labeled nucleic acids allows for the measurements of distances, solvent accessibilities and conformation dynamics. Using the spin-labeling strategy described here, spin-labeled samples can be prepared in 2-4 d. |
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Authors:
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Thomas E Edwards; Snorri Th Sigurdsson |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: Nature protocols Volume: 2 ISSN: 1750-2799 ISO Abbreviation: Nat Protoc Publication Date: 2007 |
Date Detail:
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Created Date: 2007-08-17 Completed Date: 2007-12-06 Revised Date: 2008-03-24 |
Medline Journal Info:
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Nlm Unique ID: 101284307 Medline TA: Nat Protoc Country: England |
Other Details:
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Languages: eng Pagination: 1954-62 Citation Subset: IM |
Affiliation:
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University of Iceland, Science Institute, Dunhaga 3, Reykjavik, Iceland. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Cyclic N-Oxides Electron Spin Resonance Spectroscopy / methods* Electrophoresis, Polyacrylamide Gel Nuclear Magnetic Resonance, Biomolecular RNA, Catalytic / chemistry* Spin Labels* |
| Chemical | |
Reg. No./Substance:
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0/Cyclic N-Oxides; 0/RNA, Catalytic; 0/Spin Labels; 0/hammerhead ribozyme; 2564-83-2/TEMPO |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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