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

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

Abstract

Heteronucleus-detected dipolar based correlation spectroscopy is established for assignments of 1H, 13C, and 15N resonances and structural analysis in fully protonated proteins. We demonstrate that 13C 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 1H dimension. The availability of 1H isotropic chemical shifts measured with the heteronucleus-detected fast-MAS experiments presented here is essential for the accurate determination of the 1H CSA tensors, which provide very useful structural probe. Finally, our results indicate that 13C detection in fast-MAS HETCOR experiments may be advantageous compared with 1H detection as it yields datasets of significantly higher resolution in the 13C dimension than the 1H detected HETCOR versions.



Source: Journal of Biomolecular NMR