13C-direct detected NMR experiments for the sequential J-based resonance assignment o
 

Abstract We present here a set of 13C-direct detected NMR experiments to facilitate the resonance assignment of RNA oligonucleotides. Three experiments have been developed: (1) the (H)CC-TOCSY-experiment utilizing a virtual decoupling scheme to assign the intraresidual ribose 13C-spins, (2) the (H)CPC-experiment that correlates each phosphorus with the C4â?² nuclei of adjacent nucleotides via J(C,P) couplings and (3) the (H)CPC-CCH-TOCSY-experiment that correlates the phosphorus nuclei with the respective C1â?²,H1â?² ribose signals. The experiments were applied to two RNA hairpin structures. The current set of 13C-direct detected experiments allows direct and unambiguous assignment of the majority of the hetero nuclei and the identification of the individual ribose moieties following their sequential assignment. Thus, 13C-direct detected NMR methods constitute useful complements to the conventional 1H-detected approach for the resonance assignment of oligonucleotides that is often hindered by the limited chemical shift dispersion. The developed methods can also be applied to large deuterated RNAs.

• Content Type Journal Article
• DOI 10.1007/s10858-010-9429-5
• Authors
  • Christian Richter, Johann Wolfgang Goethe-University Frankfurt Institute for Organic Chemistry and Chemical Biology, Center for Biomolecular Magnetic Resonance Max-von-Laue-Str. 7 60438 Frankfurt/M Germany
  • Helena Kovacs, Bruker BioSpin AG Industriestrasse 26 8117 Fällanden Switzerland
  • Janina Buck, Johann Wolfgang Goethe-University Frankfurt Institute for Organic Chemistry and Chemical Biology, Center for Biomolecular Magnetic Resonance Max-von-Laue-Str. 7 60438 Frankfurt/M Germany
  • Anna Wacker, Johann Wolfgang Goethe-University Frankfurt Institute for Organic Chemistry and Chemical Biology, Center for Biomolecular Magnetic Resonance Max-von-Laue-Str. 7 60438 Frankfurt/M Germany
  • Boris Fürtig, Johann Wolfgang Goethe-University Frankfurt Institute for Organic Chemistry and Chemical Biology, Center for Biomolecular Magnetic Resonance Max-von-Laue-Str. 7 60438 Frankfurt/M Germany
  • Wolfgang Bermel, Bruker BioSpin GmbH Silberstreifen 76287 Rheinstetten Germany
  • Harald Schwalbe, Johann Wolfgang Goethe-University Frankfurt Institute for Organic Chemistry and Chemical Biology, Center for Biomolecular Magnetic Resonance Max-von-Laue-Str. 7 60438 Frankfurt/M Germany

• • Journal Journal of Biomolecular NMR
  • Online ISSN 1573-5001
  • Print ISSN 0925-2738
  • Journal Volume Volume 47
  • Journal Issue Volume 47, Number 4

Source: Journal of Biomolecular NMR