Fast multidimensional NMR spectroscopy by spin-state selective off-resonance decoupling (SITAR).

Spin-state selective off-resonance decoupling (SITAR) is applied to the amide proton-to-nitrogen-to-alpha-carbon correlation (HNCA) triple-resonance experiment by measuring the 15N chemical shift during the acquisition simultaneously with the 1H chemical shift. The simultaneous detection of both 1H and 15N chemical shifts in SITAR reduces the dimensionality of the HNCA-type experiment from three dimensions to two dimensions with a 15N chemical shift resolution of approximately 0.4 ppm. This enables the recording of triple-resonance experiments in several minutes. SITAR is furthermore applied to the amide proton-to-nitrogen-to-alpha-carbon-and-beta-carbon correlation (HNCACB) triple-resonance experiment and the 15N-resolved [1H,1H]-nuclear Overhauser enhancement spectroscopy (NOESY) experiment with similar success. The accompanied peak crowding and chemical shift degeneracy of the amide protons in the SITAR two-dimensional (2D) spectra, which are inherent properties of pseudo-dimensional experiments, are resolved by local correlation of the two sub-spectra. With this procedure a 13C--1H strip for each 15N--1H moiety is generated resulting in a three-dimensional (3D) strip list known from the conventional 3D spectra. The quality of the strip list in terms of peak crowding and chemical shift degeneracy is comparable to their corresponding 3D counterparts. An analysis-software within the CARA package is presented, which generates, visualizes and manages the SITAR spectra, the corresponding strip lists and the assignment process. Copyright 2006 John Wiley & Sons, Ltd.