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Probing RNA dynamics via longitudinal exchange and CPMG relaxation dispersion NMR spectroscopy using a sensitive (13)C-methyl label
The refolding kinetics of bistable RNA sequences were studied in unperturbed equilibrium via (13)C exchange NMR spectroscopy. For this purpose a straightforward labeling technique was elaborated using a 2′-(13)C-methoxy uridine modification, which was prepared by a two-step synthesis and introduced...
Autores principales: | , , , , |
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Formato: | Texto |
Lenguaje: | English |
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Oxford University Press
2011
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3105391/ https://www.ncbi.nlm.nih.gov/pubmed/21252295 http://dx.doi.org/10.1093/nar/gkq1361 |
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author | Kloiber, Karin Spitzer, Romana Tollinger, Martin Konrat, Robert Kreutz, Christoph |
author_facet | Kloiber, Karin Spitzer, Romana Tollinger, Martin Konrat, Robert Kreutz, Christoph |
author_sort | Kloiber, Karin |
collection | PubMed |
description | The refolding kinetics of bistable RNA sequences were studied in unperturbed equilibrium via (13)C exchange NMR spectroscopy. For this purpose a straightforward labeling technique was elaborated using a 2′-(13)C-methoxy uridine modification, which was prepared by a two-step synthesis and introduced into RNA using standard protocols. Using (13)C longitudinal exchange NMR spectroscopy the refolding kinetics of a 20 nt bistable RNA were characterized at temperatures between 298 and 310 K, yielding the enthalpy and entropy differences between the conformers at equilibrium and the activation energy of the refolding process. The kinetics of a more stable 32 nt bistable RNA could be analyzed by the same approach at elevated temperatures, i.e. at 314 and 316 K. Finally, the dynamics of a multi-stable RNA able to fold into two hairpin- and a pseudo-knotted conformation was studied by (13)C relaxation dispersion NMR spectroscopy. |
format | Text |
id | pubmed-3105391 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2011 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-31053912011-06-01 Probing RNA dynamics via longitudinal exchange and CPMG relaxation dispersion NMR spectroscopy using a sensitive (13)C-methyl label Kloiber, Karin Spitzer, Romana Tollinger, Martin Konrat, Robert Kreutz, Christoph Nucleic Acids Res RNA The refolding kinetics of bistable RNA sequences were studied in unperturbed equilibrium via (13)C exchange NMR spectroscopy. For this purpose a straightforward labeling technique was elaborated using a 2′-(13)C-methoxy uridine modification, which was prepared by a two-step synthesis and introduced into RNA using standard protocols. Using (13)C longitudinal exchange NMR spectroscopy the refolding kinetics of a 20 nt bistable RNA were characterized at temperatures between 298 and 310 K, yielding the enthalpy and entropy differences between the conformers at equilibrium and the activation energy of the refolding process. The kinetics of a more stable 32 nt bistable RNA could be analyzed by the same approach at elevated temperatures, i.e. at 314 and 316 K. Finally, the dynamics of a multi-stable RNA able to fold into two hairpin- and a pseudo-knotted conformation was studied by (13)C relaxation dispersion NMR spectroscopy. Oxford University Press 2011-05 2011-01-19 /pmc/articles/PMC3105391/ /pubmed/21252295 http://dx.doi.org/10.1093/nar/gkq1361 Text en © The Author(s) 2011. Published by Oxford University Press. http://creativecommons.org/licenses/by-nc/2.5 This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.5), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | RNA Kloiber, Karin Spitzer, Romana Tollinger, Martin Konrat, Robert Kreutz, Christoph Probing RNA dynamics via longitudinal exchange and CPMG relaxation dispersion NMR spectroscopy using a sensitive (13)C-methyl label |
title | Probing RNA dynamics via longitudinal exchange and CPMG relaxation dispersion NMR spectroscopy using a sensitive (13)C-methyl label |
title_full | Probing RNA dynamics via longitudinal exchange and CPMG relaxation dispersion NMR spectroscopy using a sensitive (13)C-methyl label |
title_fullStr | Probing RNA dynamics via longitudinal exchange and CPMG relaxation dispersion NMR spectroscopy using a sensitive (13)C-methyl label |
title_full_unstemmed | Probing RNA dynamics via longitudinal exchange and CPMG relaxation dispersion NMR spectroscopy using a sensitive (13)C-methyl label |
title_short | Probing RNA dynamics via longitudinal exchange and CPMG relaxation dispersion NMR spectroscopy using a sensitive (13)C-methyl label |
title_sort | probing rna dynamics via longitudinal exchange and cpmg relaxation dispersion nmr spectroscopy using a sensitive (13)c-methyl label |
topic | RNA |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3105391/ https://www.ncbi.nlm.nih.gov/pubmed/21252295 http://dx.doi.org/10.1093/nar/gkq1361 |
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