Combining methods for speeding up multi-dimensional acquisition. Sparse sampling and fast pulsing methods for unfolded proteins

Resonance assignment of intrinsically disordered proteins is made difficult by the extensive spectral overlaps. High-resolution 3D and 4D spectra are thus essential for this purpose. We have adapted the series of 3D BEST-experiments proposed by Lescop et al. [E. Lescop, P. Schanda, B. Brutscher, A set of BEST triple-resonance experiments for time-optimized protein resonance assignment, J. Magn. Reson. 187 (2007) 163-169] to the case of unfolded proteins. Longer acquisitions in the indirect dimensions are obtained by implementing semi-constant time evolution and sparse sampling. Using maximum entropy reconstruction for the indirect dimensions, the artifact intensity due to sparse sampling can be reduced to a level similar to the other sources of noise. The reduction of the sampled increments and the shorter duration of individual transients makes it possible to record a 4D experiment with reasonable resolution in less than 60 h.