2'-Alkynyl spin-labelling is a minimally perturbing tool for DNA structural analysis
| Autor: | Tom Brown, Andrew N. Lane, Janet E. Lovett, Edward A. Anderson, Marius M. Haugland, Denis Ptchelkine, Jack S Hardwick, Frank R. Beierlein, Afaf H. El-Sagheer |
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| Přispěvatelé: | BBSRC, The Wellcome Trust, The Royal Society, University of St Andrews. Biomedical Sciences Research Complex, University of St Andrews. School of Physics and Astronomy |
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
Nitroxide mediated radical polymerization
Magnetic Resonance Spectroscopy AcademicSubjects/SCI00010 Biology Molecular Dynamics Simulation 010402 general chemistry Crystallography X-Ray 01 natural sciences law.invention 03 medical and health sciences Molecular dynamics Chemical Biology and Nucleic Acid Chemistry law Genetics QD Spectroscopy Spin (physics) Electron paramagnetic resonance 030304 developmental biology 0303 health sciences Base Sequence Oligonucleotide Electron Spin Resonance Spectroscopy DAS Nuclear magnetic resonance spectroscopy DNA QD Chemistry 0104 chemical sciences Chemical physics Nucleic acid Spin Labels |
| Zdroj: | Nucleic Acids Research |
| Popis: | Funding: Engineering and Physical Sciences Research Council [EP/M019195/1]; Engineering and Physical Sciences Research Council Studentship (to J.S.H.); Biotechnology and Biological Sciences Research Council [BB/J001694/2, BB/R021848/1]; ADTBio; University of Kentucky and NCI Cancer Center Support Grant [P30 CA177558]; The Carmen L. Buck Endowment; Emerging Fields Initiative of the Friedrich-Alexander-University of Erlangen-Nuremberg [Grant title ‘Chemistry in Live Cells’]; Wellcome Trust [099149/Z/12/Z]; Royal Society, University Research Fellowship (to J.E.L.). Funding for open access charge: University of Oxford. The determination of distances between specific points in nucleic acids is essential to understanding their behaviour at the molecular level. The ability to measure distances of 2–10 nm is particularly important: deformations arising from protein binding commonly fall within this range, but the reliable measurement of such distances for a conformational ensemble remains a significant challenge. Using several techniques, we show that electron paramagnetic resonance (EPR) spectroscopy of oligonucleotides spin-labelled with triazole-appended nitroxides at the 2′ position offers a robust and minimally perturbing tool for obtaining such measurements. For two nitroxides, we present results from EPR spectroscopy, X-ray crystal structures of B-form spin-labelled DNA duplexes, molecular dynamics simulations and nuclear magnetic resonance spectroscopy. These four methods are mutually supportive, and pinpoint the locations of the spin labels on the duplexes. In doing so, this work establishes 2′-alkynyl nitroxide spin-labelling as a minimally perturbing method for probing DNA conformation. Publisher PDF |
| Databáze: | OpenAIRE |
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