Novel three-dimensional 1H-13C-31P triple resonance experiments for sequential backbone correlations in nucleic acids.
Backbone-driven assignment methods that utilize covalent connectivities have greatly facilitated spectral assignments of proteins. In nucleic acids, 1H-13C-31P correlations could play a similar role, and several related experiments (HCP) have recently been presented for backbone-driven sequential assignments in RNA. The three-dimensional extension of 1H-31P Het-Cor (P,H-COSY-H,C-HMQC) and Het-TOCSY (P,H-TOCSY-H,C-HMQC) experiments presented here complements HCP experiments as tools for spectral assignments and extraction of dihedral angle constraints. By relying on 1H-31P rather than 13C-31P couplings to generate cross peaks, the strongest connectivities are observed in different spectral regions, increasing the likelihood of resolving spectral overlap. In addition, semiquantitative estimates of 1H-31P and 13C-31P couplings provide dihedral angle constraints for RNA structure determination.
Varani
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G
MRC Laboratory of Molecular Biology, Cambridge, U.K.
Aboul-ela
F
F
Allain
F
F
Gubser
C C
CC
D001483:Base Sequence; D002247:Carbon Isotopes; D006859:Hydrogen; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D008969:Molecular Sequence Data; D015394:Molecular Structure; D009696:Nucleic Acids / Q000737:chemistry*; D010759:Phosphorus Isotopes; D012313:RNA / Q000737:chemistry
D002247:Carbon Isotopes; D009696:Nucleic Acids; D010759:Phosphorus Isotopes; D012313:RNA; D006859:Hydrogen