Solid-Phase Chemical Synthesis of Stable Isotope-Labeled RNA to Aid Structure and Dynamics Studies by NMR Spectroscopy

Owen Becette; Lukasz T Olenginski; Theodore K Dayie

doi:10.3390/molecules24193476

Solid-Phase Chemical Synthesis of Stable Isotope-Labeled RNA to Aid Structure and Dynamics Studies by NMR Spectroscopy

Molecules. 2019 Sep 25;24(19):3476. doi: 10.3390/molecules24193476.

Authors

Owen Becette¹, Lukasz T Olenginski², Theodore K Dayie³

Affiliations

¹ Center for Biomolecular Structure and Organization, Department of Chemistry and Biochemistry, University of Maryland, College Park, MD 20742, USA. obecette@umd.edu.
² Center for Biomolecular Structure and Organization, Department of Chemistry and Biochemistry, University of Maryland, College Park, MD 20742, USA. lolengin@umd.edu.
³ Center for Biomolecular Structure and Organization, Department of Chemistry and Biochemistry, University of Maryland, College Park, MD 20742, USA. dayie@umd.edu.

Abstract

RNA structure and dynamic studies by NMR spectroscopy suffer from chemical shift overlap and line broadening, both of which become worse as RNA size increases. Incorporation of stable isotope labels into RNA has provided several solutions to these limitations. Nevertheless, the only method to circumvent the problem of spectral overlap completely is the solid-phase chemical synthesis of RNA with labeled RNA phosphoramidites. In this review, we summarize the practical aspects of this methodology for NMR spectroscopy studies of RNA. These types of investigations lie at the intersection of chemistry and biophysics and highlight the need for collaborative efforts to tackle the integrative structural biology problems that exist in the RNA world. Finally, examples of RNA structure and dynamic studies using labeled phosphoramidites are highlighted.

Keywords: NMR spectroscopy; RNA; dynamics; phosphoramidite; solid-phase synthesis; stable isotope; structure.

Publication types

Review

MeSH terms

Isotope Labeling*
Molecular Dynamics Simulation*
Molecular Structure
Nuclear Magnetic Resonance, Biomolecular
Nucleic Acid Conformation*
RNA / chemical synthesis
RNA / chemistry*
Solid-Phase Synthesis Techniques*

Substances

RNA

Grants and funding

1808705/National Science Foundation