caa a22 4500 445325410 CHVBK 20180317142718.0 cr unu---uuuuu 170323e20110901xx s 000 0 eng 10.1007/s10858-011-9544-y doi (NATIONALLICENCE)springer-10.1007/s10858-011-9544-y Measurement of 1H-15N and 1H-13C residual dipolar couplings in nucleic acids from TROSY intensities [Elektronische Daten] [Jinfa Ying, Jinbu Wang, Alex Grishaev, Ping Yu, Yun-Xing Wang, Ad Bax] Analogous to the recently introduced ARTSY method for measurement of one-bond 1H-15N residual dipolar couplings (RDCs) in large perdeuterated proteins, we introduce methods for measurement of base 13C-1H and 15N-1H RDCs in protonated nucleic acids. Measurements are based on quantitative analysis of intensities in 1H-15N and 13C-1H TROSY-HSQC spectra, and are illustrated for a 71-nucleotide adenine riboswitch. Results compare favorably with those of conventional frequency-based measurements in terms of completeness and convenience of use. The ARTSY method derives the size of the coupling from the ratio of intensities observed in two TROSY-HSQC spectra recorded with different dephasing delays, thereby minimizing potential resonance overlap problems. Precision of the RDC measurements is limited by the signal-to-noise ratio, S/N, achievable in the 2D TROSY-HSQC reference spectrum, and is approximately given by 30/(S/N) Hz for 15N-1H and 65/(S/N) Hz for 13C-1H. The signal-to-noise ratio of both 1H-15N and 1H-13C spectra greatly benefits when water magnetization during the experiments is not perturbed, such that rapid magnetization transfer from bulk water to the nucleic acid, mediated by rapid amino and hydroxyl hydrogen exchange coupled with 1H-1H NOE transfer, allows for fast repetition of the experiment. RDCs in the mutated helix 1 of the riboswitch are compatible with nucleotide-specifically modeled, idealized A-form geometry and a static orientation relative to the helix 2/3 pair, which differs by ca 6° relative to the X-ray structure of the native riboswitch. Springer Science+Business Media B.V. (outside the USA), 2011 ARTSY nationallicence BEST nationallicence A-form RNA nationallicence Quantitative J correlation nationallicence TROSY nationallicence RDC nationallicence RNA nationallicence DNA nationallicence Ying Jinfa Laboratory of Chemical Physics, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Building 5, Room 126, 20892-0520, Bethesda, MD, USA aut Wang Jinbu Structural Biophysics Laboratory, National Cancer Institute, National Institutes of Health, 21702, Frederick, MD, USA aut Grishaev Alex Laboratory of Chemical Physics, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Building 5, Room 126, 20892-0520, Bethesda, MD, USA aut Yu Ping Structural Biophysics Laboratory, National Cancer Institute, National Institutes of Health, 21702, Frederick, MD, USA aut Wang Yun-Xing Structural Biophysics Laboratory, National Cancer Institute, National Institutes of Health, 21702, Frederick, MD, USA aut Bax Ad Laboratory of Chemical Physics, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Building 5, Room 126, 20892-0520, Bethesda, MD, USA aut Journal of Biomolecular NMR Springer Netherlands 51/1-2(2011-09-01), 89-103 0925-2738 51:1-2<89 2011 51 10858 https://doi.org/10.1007/s10858-011-9544-y text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s10858-011-9544-y text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Ying Jinfa Laboratory of Chemical Physics, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Building 5, Room 126, 20892-0520, Bethesda, MD, USA aut NATIONALLICENCE 700 1- Wang Jinbu Structural Biophysics Laboratory, National Cancer Institute, National Institutes of Health, 21702, Frederick, MD, USA aut NATIONALLICENCE 700 1- Grishaev Alex Laboratory of Chemical Physics, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Building 5, Room 126, 20892-0520, Bethesda, MD, USA aut NATIONALLICENCE 700 1- Yu Ping Structural Biophysics Laboratory, National Cancer Institute, National Institutes of Health, 21702, Frederick, MD, USA aut NATIONALLICENCE 700 1- Wang Yun-Xing Structural Biophysics Laboratory, National Cancer Institute, National Institutes of Health, 21702, Frederick, MD, USA aut NATIONALLICENCE 700 1- Bax Ad Laboratory of Chemical Physics, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Building 5, Room 126, 20892-0520, Bethesda, MD, USA aut NATIONALLICENCE 773 0- Journal of Biomolecular NMR Springer Netherlands 51/1-2(2011-09-01), 89-103 0925-2738 51:1-2<89 2011 51 10858 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer