An Approach to the Structure Determination of Larger Proteins Using Triple Resonance NMR Experiments in Conjunction with Random Fractional Deuteration

Abstract

A combination of simulation and experiment is used to demonstrate that the sensitivity of a family of 3D/4D NMR experiments used to assign resonances and to obtain structural restraints in proteins is improved by partial random deuteration; the improvement increases as the correlation time of the protein becomes longer. The results suggest that deuteration at a level of ∼50% optimizes the sensitivity of experiments which are used to assign sidechain 1H and 13C resonances by correlating them with the resonances from backbone nuclei. In addition, this level of deuteration is also a good compromise for recording NOESY experiments. Using this approach, it should be possible to determine structures of larger proteins.