2020
DOI: 10.1002/anie.202006577
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Aromatic 19F–13C TROSY—[19F, 13C]‐Pyrimidine Labeling for NMR Spectroscopy of RNA

Abstract: We present the access to [5‐19F, 5‐13C]‐uridine and ‐cytidine phosphoramidites for the production of site‐specifically modified RNAs up to 65 nucleotides (nts). The amidites were used to introduce [5‐19F, 5‐13C]‐pyrimidine labels into five RNAs—the 30 nt human immunodeficiency virus trans activation response (HIV TAR) 2 RNA, the 61 nt human hepatitis B virus ϵ (hHBV ϵ) RNA, the 49 nt SAM VI riboswitch aptamer domain from B. angulatum, the 29 nt apical stem loop of the pre‐microRNA (miRNA) 21 and the 59 nt full… Show more

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Cited by 24 publications
(18 citation statements)
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References 49 publications
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“…In general, 19 F based experiment can thus be expected to outperform more traditional measurements at the elevated temperatures or at a higher pH. Finally, recent advances in labeling techniques have made RNA and DNA accessible for 19 F NMR (Sochor et al 2016;Nußbaumer et al 2020;Baranowski et al 2020), thereby further expanding the applicability of the presented methodologies. In summary, our approach will strengthen the use of 19 F NMR to accurately quantify dynamic processes in a broad range of biological systems.…”
Section: Discussionmentioning
confidence: 99%
“…In general, 19 F based experiment can thus be expected to outperform more traditional measurements at the elevated temperatures or at a higher pH. Finally, recent advances in labeling techniques have made RNA and DNA accessible for 19 F NMR (Sochor et al 2016;Nußbaumer et al 2020;Baranowski et al 2020), thereby further expanding the applicability of the presented methodologies. In summary, our approach will strengthen the use of 19 F NMR to accurately quantify dynamic processes in a broad range of biological systems.…”
Section: Discussionmentioning
confidence: 99%
“…19 However since that time, Aryl 19 F- 13 C TROSY methods have been reported for biopolymers, 36 and have now been applied to ligand binding. 50 However, only a limited set of fluorinated amino acids and nucleic acids have been used, in part due to the requirement of specific 13 C and 19 F labeling. Improved access to these building blocks may augment the types of biopolymers which can be studied by this approach.…”
Section: F Iloe Nmr Experiments Discussedmentioning
confidence: 99%
“… 36 This has led to the characterization of proteins as large as 180 kDa, and has more recently been applied to 13 C-labeled fluorouracil in RNA including ligand binding studies. 50 Conversely, CF 3 -TROSY may be limited to lower field strengths where the CSA effects are negligible. At high field strengths, TROSY effects were not observed due to the high CSA of the CF 3 group.…”
Section: Introductionmentioning
confidence: 99%
“…Even with these innovations, spectral crowding and signal overlap still limit the effective analysis of large (> 30 nucleotide, nt) RNAs [18]. To tackle this problem, we and others have developed labeling technologies to incorporate stable isotopes (i.e., 2 H, 13 C, 19 F, and 15 N) into RNA to benefit NMR structure and dynamics measurements [7,[19][20][21][22][23][24][25][26][27].…”
Section: Introductionmentioning
confidence: 99%