TEDOR with adiabatic inversion pulses: Resonance assignments of 13C/15N labelled RNAs

J Biomol NMR. 2005 Jan;31(1):49-57. doi: 10.1007/s10858-004-6066-x.

Abstract

We have examined via numerical simulations the performance characteristics of different 15N RF pulse schemes employed in the transferred echo double resonance (TEDOR) experimental protocol for generating 13C-15N dipolar chemical shift correlation spectra of isotopically labelled biological systems at moderate MAS frequencies (omega(r) approximately 10 kHz). With an 15N field strength of approximately 30-35 kHz that is typically available in 5 mm triple resonance MAS NMR probes, it is shown that a robust TEDOR sequence with significant tolerance to experimental imperfections sa as H1 inhomogeneity and resonance offsets can be effectively implemented using adiabatic heteronuclear dipolar recoupling pulse schemes. TEDOR-based 15N-13C and 15N-13C-13C chemical shift correlation experiments were carried out for obtaining 13C and 15N resonance assignments of an RNA composed of 97 (CUG) repeats which has been implicated in the neuromuscular disease myotonic dystrophy.

MeSH terms

  • Carbon Isotopes
  • Magnetic Resonance Spectroscopy / methods*
  • Nitrogen Isotopes
  • RNA / chemistry*
  • Time Factors

Substances

  • Carbon Isotopes
  • Nitrogen Isotopes
  • RNA