Simultaneous H-1- or H-2-, N-15- and multiple-band-selective C-13-decoupling during acquisition in C-13-detected experiments with proteins and oligonucleotides

Significant resolution improvement in C-13,C-13-TOCSY spectra of uniformly deuterated and C-13,N-15-labeled protein and C-13,N-15-labeled RNA samples is achieved by introduction of multiple-band-selective C-13-homodecoupling applied simultaneously with H-1- or H-2- and N-15-decoupling at all stages of multidimensional experiments including signal acquisition period. The application of single, double or triple band-selective C-13-decoupling in 2D-[C-13,C-13]-TOCSY experiments during acquisition strongly simplifies the homonuclear splitting pattern. The technical aspects of complex multiple-band homonuclear decoupling and hardware requirements are discussed. The use of this technique (i) facilitates the resonance assignment process as it reduces signal overlap in homonuclear C-13-spectra and (ii) possibly improves the signal-to-noise ratio through multiplet collapse. It can be applied in any C-13-detected experiment.