Base-type-selective high-resolution 13C edited NOESY for sequential assignment of large RNAs

J Biomol NMR. 2001 Feb;19(2):141-51. doi: 10.1023/a:1008340210079.

Abstract

Extensive spectral overlap presents a major problem for the NMR study of large RNAs. Here we present NMR techniques for resolution enhancement and spectral simplification of fully 13C labelled RNA. High-resolution 1H-13C correlation spectra are obtained by combining TROSY-type experiments with multiple-band-selective homonuclear 13C decoupling. An additional C-C filter sequence performs base-type-selective spectral editing. Signal loss during the filter is significantly reduced because of TROSY-type spin evolution. These tools can be inserted in any 13C-edited multidimensional NMR experiment. As an example we have chosen the 13C-edited NOESY which is a crucial experiment for sequential resonance assignment of RNA. Application to a 33-nucleotide RNA aptamer and a 76-nucleotide tRNA illustrates the potential of this new methodology.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Carbon Isotopes
  • Hydrogen / chemistry
  • Magnetic Resonance Spectroscopy / methods*
  • Models, Molecular
  • Molecular Sequence Data
  • Nucleic Acid Conformation
  • RNA / chemistry*
  • RNA, Transfer / chemistry

Substances

  • Carbon Isotopes
  • RNA
  • Hydrogen
  • RNA, Transfer