NDLI: Aromatic$ ^{19}$ F–$ ^{13}$ C TROSY—[$ ^{19}$ F,$ ^{13}$ C]‐Pyrimidine Labeling for NMR Spectroscopy of RNA

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Aromatic$ ^{19}$ F–$ ^{13}$ C TROSY—[$ ^{19}$ F,$ ^{13}$ C]‐Pyrimidine Labeling for NMR Spectroscopy of RNA

Content Provider	Scilit
Author	Nußbaumer, Felix Plangger, Raphael Roeck, Manuel Kreutz, Christoph
Copyright Year	2020
Description	Journal: Angewandte Chemie International Edition 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 30nt human immunodeficiency virus trans activation response (HIV TAR) 2 RNA, the 61nt human hepatitis B virus ε (hHBV ε) RNA, the 49nt SAM VI riboswitch aptamer domain from B. angulatum, the 29nt apical stem loop of the pre‐microRNA (miR) 21 and the 59nt full length pre‐miR 21. The main stimulus to introduce the aromatic 19F‐13C‐spin topology into RNA comes from a work of Boeszoermenyi et al., in which the dipole‐dipole interaction and the chemical shift anisotropy relaxation mechanisms cancel each other leading to advantageous TROSY properties shown for aromatic protein sidechains. This aromatic 13C‐19F labeling scheme is now transferred to RNA. We provide a protocol for the resonance assignment by solid phase synthesis based on diluted [5‐19F, 5‐13C]/[5‐19F] pyrimidine labeling. For the 61nt hHBV ε we find a beneficial 19F‐13C TROSY enhancement, which should be even more pronounced in larger RNAs and will facilitate the NMR studies of larger RNAs. The [19F, 13C]‐labeling of the SAM VI aptamer domain and the pre‐miR 21 further opens the possibility to use the biorthogonal stable isotope reporter nuclei in in vivo NMR to observe ligand binding and microRNA processing in a biological relevant setting.
Related Links	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7540360/pdf
Ending Page	17069
Page Count	8
Starting Page	17062
ISSN	14337851
e-ISSN	15213773
DOI	10.1002/anie.202006577
Journal	Angewandte Chemie International Edition
Issue Number	39
Volume Number	59
Language	English
Publisher	Wiley-Blackwell
Publisher Date	2020-06-18
Access Restriction	Open
Subject Keyword	Journal: Angewandte Chemie International Edition Stable Isotope Labeling
Content Type	Text
Resource Type	Article
Subject	Chemistry Catalysis

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