Simple and Robust Study of Backbone Dynamics of Crystalline Proteins Employing ¹H-¹⁵N Dipolar Coupling Dispersion

We report a new solid-state multidimensional NMR approach based on the cross-polarization with variable-contact pulse sequence [ Paluch , P. ; Pawlak , T. ; Amoureux , J.-P. ; Potrzebowski , M. J. J. Magn. Reson. 233 , 2013 , 56 ], with ¹H inverse detection and very fast magic angle spinning (ν_R = 60 kHz), dedicated to the measurement of local molecular motions of ¹H-¹⁵N vectors. The introduced three-dimensional experiments, ¹H-¹⁵N-¹H and hCA(N)H, are particularly useful for the study of molecular dynamics of proteins and other complex structures. The applicability and power of this methodology have been revealed by employing as a model sample the GB-1 small protein doped with Na₂CuEDTA. The results clearly prove that the dispersion of ¹H-¹⁵N dipolar coupling constants well correlates with higher order structure of the protein. Our approach complements the conventional studies and offers a fast and reasonably simple method.