Abstract
Protein motions over various time scales are crucial for protein function. NMR relaxation dispersion experiments play a key role in explaining these motions. However, the study of slow conformational changes with lowly populated states remained elusive. The recently developed exchange-mediated saturation transfer experiments allow the detection and characterization of such motions, but require extensive measurement time. Here we show that, by making use of Fourier transform, the total acquisition time required to measure an exchange-mediated saturation transfer profile can be reduced by twofold in case that one applies linear prediction. In addition, we demonstrate that the analytical solution for R1ρ experiments can be used for fitting the exchange-mediated saturation transfer profile. Furthermore, we show that simultaneous analysis of exchange-mediated saturation transfer profiles with two different radio-frequency field strengths is required for accurate and precise characterization of the exchange process and the exchanging states.
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Acknowledgments
The authors thank L. M. I. Koharudin and A. M. Gronenborn for kindly providing 15N-labeled OAA sample. We also thank G. Bouvignies and L. E. Kay for providing a software to validate our in-house software for fitting CEST profiles. This work was supported by the James Graham Brown Foundation, the Max Planck Society and the EU (ERC Grant Agreement Number 233227 to CG).
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Carneiro, M.G., Reddy, J.G., Griesinger, C. et al. Speeding-up exchange-mediated saturation transfer experiments by Fourier transform. J Biomol NMR 63, 237–244 (2015). https://doi.org/10.1007/s10858-015-9985-9
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DOI: https://doi.org/10.1007/s10858-015-9985-9