Journal of Biomolecular NMR

, Volume 57, Issue 1, pp 1–9 | Cite as

4D Non-uniformly sampled C,C-NOESY experiment for sequential assignment of 13C,15N-labeled RNAs

  • Jan Stanek
  • Peter Podbevšek
  • Wiktor Koźmiński
  • Janez Plavec
  • Mirko Cevec
Communication

Abstract

A 4D 13C(aromatic),13C(ribose)-edited NOESY experiment is introduced to improve sequential assignment of non-coding RNA, often hampered by a limited dispersion of 1H and 13C chemical shifts. The 13C-labeling of RNA is fully utilized by inclusion of two 13C evolution periods. These dimensions provide enhanced dispersion of resonances in the 4D spectrum. High spectral resolution is obtained using random non-uniform sampling in three indirect dimensions. The autocorrelation peaks are efficiently suppressed using band-selective pulses. Since the dynamic range of observed resonances is significantly decreased, the reconstruction of the 4D spectrum is greatly simplified. The experiment can replace two conventionally sampled 3D NOESY spectra (either ribose-13C- or aromatic-13C-separated), and remove most ambiguities encountered during sequential walks. The assignment strategy based on a homonuclear and 4D C,C-edited NOESY experiments is proposed and verified on a 34-nt RNA showing typical structure elements.

Keywords

Multidimensional NMR Non-uniform sampling Isotope labeled RNA Resonance assignment 

Supplementary material

10858_2013_9771_MOESM1_ESM.pdf (600 kb)
Supplementary material 1 (PDF 600 kb)

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Copyright information

© Springer Science+Business Media Dordrecht 2013

Authors and Affiliations

  • Jan Stanek
    • 1
  • Peter Podbevšek
    • 2
  • Wiktor Koźmiński
    • 1
  • Janez Plavec
    • 2
    • 3
    • 4
  • Mirko Cevec
    • 2
  1. 1.Faculty of Chemistry, Biological and Chemical Research CentreUniversity of WarsawWarszawaPoland
  2. 2.Slovenian NMR CentreNational Institute of ChemistryLjubljanaSlovenia
  3. 3.EN-FIST Centre of ExcellenceLjubljanaSlovenia
  4. 4.Faculty of Chemistry and Chemical TechnologyUniversity of LjubljanaLjubljanaSlovenia

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