Abstract
Recent methyl adiabatic relaxation dispersion experiments provide examination of conformational dynamics across a very wide timescale (102–105 s−1) and, particularly, provide insight into the hydrophobic core of proteins and allosteric effects associated with modulators. The experiments require efficient decoupling of 1H and 13C spin interactions, and some artifacts have been discovered, which are associated with the design of the proton decoupling scheme. The experimental data suggest that the original design is valid; however, pulse sequences with either no proton decoupling or proton decoupling with imperfect pulses can potentially exhibit complications in the experiments. Here, we demonstrate that pulse imperfections in the proton decoupling scheme can be dramatically alleviated by using a single composite π pulse and provide pure single-exponential relaxation data. It allows the opportunity to access high-quality methyl adiabatic relaxation dispersion data by removing the cross-correlation between dipole–dipole interaction and chemical shift anisotropy. The resulting high-quality data is illustrated with the binding of an allosteric modulator (G2BR) to the ubiquitin conjugating enzyme Ube2g2.
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Acknowledgements
The authors gratefully acknowledge the use of the Biophysics Resource, Structural Biophysics Laboratory, and the assistance of Dr. Sergey Tarasov and Ms. Marzena Dyba. The research was supported by the Intramural Research Program of the National Cancer Institute, Projects ZIA BC 011131 and ZIA BC 011132. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.
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Chao, FA., Khago, D. & Byrd, R.A. Achieving pure spin effects by artifact suppression in methyl adiabatic relaxation experiments. J Biomol NMR 74, 223–228 (2020). https://doi.org/10.1007/s10858-020-00312-2
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DOI: https://doi.org/10.1007/s10858-020-00312-2