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

Authors
  • Stanek, Jan1
  • Podbevšek, Peter2
  • Koźmiński, Wiktor1
  • Plavec, Janez2, 3, 4
  • Cevec, Mirko2
  • 1 University of Warsaw, Faculty of Chemistry, Biological and Chemical Research Centre, Żwirki i Wigury 101, Warszawa, 02-089, Poland , Warszawa (Poland)
  • 2 National Institute of Chemistry, Slovenian NMR Centre, Hajdrihova ulica 19, Ljubljana, 1000, Slovenia , Ljubljana (Slovenia)
  • 3 EN-FIST Centre of Excellence, Dunajska cesta 156, Ljubljana, 1000, Slovenia , Ljubljana (Slovenia)
  • 4 University of Ljubljana, Faculty of Chemistry and Chemical Technology, Aškerčeva cesta 5, Ljubljana, 1000, Slovenia , Ljubljana (Slovenia)
Type
Published Article
Journal
Journal of Biomolecular NMR
Publisher
Springer-Verlag
Publication Date
Aug 21, 2013
Volume
57
Issue
1
Pages
1–9
Identifiers
DOI: 10.1007/s10858-013-9771-5
Source
Springer Nature
Keywords
License
Yellow

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.

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