Dynamics-Function Analysis in Catalytic RNA Using NMR Spin Relaxation and Conformationally Restricted Nucleotides

Methods Mol Biol. 2021:2167:183-202. doi: 10.1007/978-1-0716-0716-9_11.

Abstract

A full understanding of biomolecular function requires an analysis of both the dynamic properties of the system of interest and the identification of those dynamics that are required for function. We describe NMR methods based on metabolically directed specific isotope labeling for the identification of molecular disorder and/or conformational transitions on the RNA backbone ribose groups. These analyses are complemented by the use of synthetic covalently modified nucleotides constrained to a single sugar pucker, which allow functional assessment of dynamics by selectively removing a minor conformer identified by NMR from the structural ensemble.

Keywords: Conformational restriction; Dynamics-function; NMR dynamics; Noncoding RNA; Sugar pucker.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Carbon Isotopes / chemistry
  • Enzyme Assays / methods*
  • Kinetics
  • Nuclear Magnetic Resonance, Biomolecular / methods*
  • Nucleic Acid Conformation*
  • Nucleotides
  • Quantitative Structure-Activity Relationship
  • RNA, Catalytic / chemistry*
  • RNA, Catalytic / genetics*
  • RNA, Untranslated / chemistry
  • RNA, Untranslated / genetics
  • Ribose / chemistry*

Substances

  • Carbon Isotopes
  • Nucleotides
  • RNA, Catalytic
  • RNA, Untranslated
  • Ribose