Fast magic angle spinning NMR with heteronucleus detection for resonance assignments and structural characterization of fully protonated proteins

Heteronucleus-detected dipolar based correlation spectroscopy is established for assignments of H-1, C-13, and N-15 resonances and structural analysis in fully protonated proteins. We demonstrate that C-13 detected 3D experiments are highly efficient and permit assignments of the majority of backbone resonances, as shown in an 89-residue dynein light chain 8, LC8 protein. With these experiments, we have resolved many ambiguities that were persistent in our previous studies using moderate MAS frequencies and lacking the H-1 dimension. The availability of H-1 isotropic chemical shifts measured with the heteronucleus-detected fast-MAS experiments presented here is essential for the accurate determination of the H-1 CSA tensors, which provide very useful structural probe. Finally, our results indicate that C-13 detection in fast-MAS HETCOR experiments may be advantageous compared with H-1 detection as it yields datasets of significantly higher resolution in the C-13 dimension than the H-1 detected HETCOR versions.