Sequential assignments in uniformly 13C- and 15N-labelled RNAs: the HC(N,P) and HC(N,P)-CCH-TOCSY experiments

J Biomol NMR. 1996 May;7(3):251-5. doi: 10.1007/BF00202042.

Abstract

An approach for the simultaneous acquisition of HCN and HCP as well as HCN-CCH-TOCSY and HCP-CCH-TOCSY triple resonance data sets for 13C-/15N-labelled RNAs is presented. The new HCN-CCH-TOCSY scheme unambiguously links all sugar resonances to the base nitrogen. In addition, simultaneous acquisition of HCN-CCH-TOCSY and HCP-CCH-TOCSY data sets provides sequential and base-type information in a single experiment, thereby saving data acquisition time as well as providing complementary data sets that are useful in clarifying ambiguous assignments. Virtually complete sequence-specific phosphate-ribose 1H, 31P, and base 15N1,9, assignments as well as partial 13C assignments could be obtained in a single experiment for a 0.5-mM sample of a 19-mer ribonucleotide.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Carbon Isotopes
  • Data Interpretation, Statistical
  • Magnetic Resonance Spectroscopy / methods*
  • Molecular Sequence Data
  • Molecular Structure
  • Nitrogen Isotopes
  • RNA / chemistry*

Substances

  • Carbon Isotopes
  • Nitrogen Isotopes
  • RNA