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Chemo-enzymatic synthesis of 13C- and 19F-labeled uridine-5′-triphosphate for RNA NMR probing

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Abstract

Solution NMR of the magnetically active fluorine-19 (19F) isotope offers tremendous advantages in the study of RNA structure and conformational changes. However, exploiting these advantages requires the synthesis of fluorinated RNA-building blocks, and this comes with certain challenges in overall yield of nucleotides, purity, and cost. Here, we detail a scalable and reliable method to produce [1′,5-13C2, 5-19F, 6-2H]- and [1′,5-13C2, 5-19F, 6-2H, 1,3-15N2]-5-fluorouracil-5′-triphosphate. We further present the steps employed to characterize the labeled nucleotide before its incorporation into RNAs with relevant biological activities.

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Acknowledgements

We are grateful to Professors Philip Deshong, and Lai-Xi Wang (University of Maryland) for use of laboratory space, and Prof Jeff Davis for many useful discussions and comments. We also thank Maryia Svirydava for her help in analyzing samples by mass spectrometry.

Funding

The funding was received by National Institute of General Medical Sciences (Grant no. U54AI50470).

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Author notes

  1. Kehinde M. Taiwo and Owen B. Becette contributed equally.

Authors and Affiliations

  1. Center for Biomolecular Structure and Organization, Department of Chemistry and Biochemistry, University of Maryland, College Park, MD, 20742, USA

    Kehinde M. Taiwo, Owen B. Becette, Guanghui Zong, Bin Chen, Peter Y. Zavalij & Theodore Kwaku Dayie

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  1. Kehinde M. Taiwo
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  2. Owen B. Becette
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  3. Guanghui Zong
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  4. Bin Chen
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  6. Theodore Kwaku Dayie
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Correspondence to Theodore Kwaku Dayie.

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Taiwo, K.M., Becette, O.B., Zong, G. et al. Chemo-enzymatic synthesis of 13C- and 19F-labeled uridine-5′-triphosphate for RNA NMR probing. Monatsh Chem 152, 441–447 (2021). https://doi.org/10.1007/s00706-021-02757-z

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  • DOI: https://doi.org/10.1007/s00706-021-02757-z

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