Measurement of protein backbone (CO)-C-13 and N-15 relaxation dispersion at high resolution

• Published in: Journal of biomolecular NMR 2017, Vol.69 (1), p.1
• Main Authors: Mayzel, Maxim, Ahlner, Alexandra, Lundström, Patrik, Orekhov, Vladislav Y.
• Format: Article
• Language: English
• Subjects: Biochemistry & Molecular Biology; Biochemistry and Molecular Biology; Biokemi och molekylärbiologi; chemical-shifts; Conformational exchange; decomposition; Dynamics; Fysikalisk kemi; IDP; invisible excited-states; multidimensional nmr; NMR; nmr-spectroscopy; NUS; Physical Chemistry; pulses; sequence; Spectroscopy; Target acquisition
• ISSN: 1573-5001; 0925-2738
• DOI: 10.1007/s10858-017-0127-4

Peak overlap in crowded regions of two-dimensional spectra prevents characterization of dynamics for many sites of interest in globular and intrinsically disordered proteins. We present new three-dimensional pulse sequences for measurement of Carr-Purcell-Meiboom-Gill relaxation dispersions at backbone nitrogen and carbonyl positions. To alleviate increase in the measurement time associated with the additional spectral dimension, we use non-uniform sampling in combination with two distinct methods of spectrum reconstruction: compressed sensing and co-processing with multi-dimensional decomposition. The new methodology was validated using disordered protein CD79A from B-cell receptor and an SH3 domain from Abp1p in exchange between its free form and bound to a peptide from the protein Ark1p. We show that, while providing much better resolution, the 3D NUS experiments give the similar accuracy and precision of the dynamic parameters to ones obtained using traditional 2D experiments. Furthermore, we show that jackknife resampling of the spectra yields robust estimates of peak intensities errors, eliminating the need for recording duplicate data points.