Synthesis of [7-15N]-GTPs for RNA structure and dynamics by NMR spectroscopy

Kehinde M Taiwo; Lukasz T Olenginski; Felix Nußbaumer; Hyeyeon Nam; Stefan Hilber; Christoph Kreutz; T Kwaku Dayie

doi:10.1007/s00706-022-02892-1

Synthesis of [7-¹⁵N]-GTPs for RNA structure and dynamics by NMR spectroscopy

Monatsh Chem. 2022;153(3):293-299. doi: 10.1007/s00706-022-02892-1. Epub 2022 Feb 26.

Authors

Kehinde M Taiwo¹, Lukasz T Olenginski¹, Felix Nußbaumer², Hyeyeon Nam^{1

3}, Stefan Hilber², Christoph Kreutz², T Kwaku Dayie¹

Affiliations

¹ Department of Chemistry and Biochemistry, Center for Biomolecular Structure and Organization, University of Maryland, College Park, MD 20742 USA.
² Institute of Organic Chemistry and Center for Molecular Biosciences Innsbruck, University of Innsbruck, Innrain 80/82, 6020 Innsbruck, Austria.
³ Present Address: Center for Cancer Research, National Cancer Institute, Frederick, MD 21702 USA.

Abstract

Several isotope-labeling strategies have been developed for the study of RNA by nuclear magnetic resonance (NMR) spectroscopy. Here, we report a combined chemical and enzymatic synthesis of [7-¹⁵N]-guanosine-5'-triphosphates for incorporation into RNA via T7 RNA polymerase-based in vitro transcription. We showcase the utility of these labels to probe both structure and dynamics in two biologically important RNAs.

Supplementary information: The online version contains supplementary material available at 10.1007/s00706-022-02892-1.

Keywords: Dynamics; Isotopic labeling; Nucleic acids; Spectroscopy.

Abstract

Grants and funding