Juen, MA;
Wunderlich, CH;
Nussbaumer, F;
Tollinger, M;
Kontaxis, G;
Konrat, R;
Hansen, DF;
(2016)
Excited States of Nucleic Acids Probed by Proton Relaxation Dispersion NMR Spectroscopy.
Angewandte Chemie International Edition
, 55
(39)
pp. 12008-12012.
10.1002/anie.201605870.
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Abstract
In this work an improved stable isotope labeling protocol for nucleic acids is introduced. The novel building blocks eliminate/minimize homonuclear 13C and 1H scalar couplings thus allowing proton relaxation dispersion (RD) experiments to report accurately on the chemical exchange of nucleic acids. Using site-specific 2H and 13C labeling, spin topologies are introduced into DNA and RNA that make 1H relaxation dispersion experiments applicable in a straightforward manner. The novel RNA/DNA building blocks were successfully incorporated into two nucleic acids. The A-site RNA was previously shown to undergo a two site exchange process in the micro- to millisecond time regime. Using proton relaxation dispersion experiments the exchange parameters determined earlier could be recapitulated, thus validating the proposed approach. We further investigated the dynamics of the cTAR DNA, a DNA transcript that is involved in the viral replication cycle of HIV-1. Again, an exchange process could be characterized and quantified. This shows the general applicablility of the novel labeling scheme for 1H RD experiments of nucleic acids.
| Type: | Article |
|---|---|
| Title: | Excited States of Nucleic Acids Probed by Proton Relaxation Dispersion NMR Spectroscopy |
| Open access status: | An open access version is available from UCL Discovery |
| DOI: | 10.1002/anie.201605870 |
| Publisher version: | http://dx.doi.org/10.1002/anie.201605870 |
| Language: | English |
| Additional information: | © 2016 The Authors. Published by Wiley-VCH Verlag GmbH & Co. KGaA. This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. |
| Keywords: | Science & Technology, Physical Sciences, Chemistry, Multidisciplinary, Chemistry, DNA, NMR spectroscopy, proton relaxation dispersion, RNA, stable isotope labeling, HIV-1 NUCLEOCAPSID PROTEIN, H-1-NMR CHEMICAL-SHIFTS, RNA DYNAMICS, CTAR DNA, Conformational States, H-1, Time, Nucleotides, Recognition, Chaperone |
| UCL classification: | UCL UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > Div of Biosciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > Div of Biosciences > Structural and Molecular Biology |
| URI: | https://discovery.ucl.ac.uk/id/eprint/1516783 |
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