Exact Distances and Internal Dynamics of Perdeuterated Ubiquitin from NOE Buildups

It is proposed to convert nuclear Overhauser effects (NOEs) into relatively precise distances for detailed structural studies of proteins. To this purpose, it is demonstrated that the measurement of NOE buildups between amide protons in perdeuterated human ubiquitin using a designed 15N-resolved HMQC-NOESY experiment enables the determination of 1HN−1HN distances up to 5 A with high accuracy and precision. These NOE-derived distances have an experimental random error of ∼0.07 A, which is smaller than the pairwise rmsd (root-mean-square deviation) of 0.24 A obtained with corresponding distances extracted from either an NMR or an X-ray structure (pdb codes: 1D3Z and 1UBQ), and also smaller than the pairwise rmsd between distances from X-ray and NMR structures (0.15 A). Because the NOE contains both structural and dynamical information, a comparison between the 3D structures and NOE-derived distances may also give insights into through-space dynamics. It appears that the extraction of motional information fr...