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Enhanced accuracy of kinetic information from CT-CPMG experiments by transverse rotating-frame spectroscopy.

MPS-Authors
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Ban,  D.
Department of NMR-based Structural Biology, MPI for biophysical chemistry, Max Planck Society;

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Mazur,  A.
Department of NMR-based Structural Biology, MPI for biophysical chemistry, Max Planck Society;

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Giao Carneiro,  M.
Department of NMR-based Structural Biology, MPI for biophysical chemistry, Max Planck Society;

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Sabo,  T. M.
Department of NMR-based Structural Biology, MPI for biophysical chemistry, Max Planck Society;

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Giller,  K.
Department of NMR-based Structural Biology, MPI for biophysical chemistry, Max Planck Society;

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Becker,  S.
Department of NMR-based Structural Biology, MPI for biophysical chemistry, Max Planck Society;

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Griesinger,  C.
Department of NMR-based Structural Biology, MPI for biophysical chemistry, Max Planck Society;

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Lee,  D.
Department of NMR-based Structural Biology, MPI for biophysical chemistry, Max Planck Society;

Fulltext (public)

1836786.pdf
(Publisher version), 656KB

Supplementary Material (public)
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Citation

Ban, D., Mazur, A., Giao Carneiro, M., Sabo, T. M., Giller, K., Koharudin, L. M. I., et al. (2013). Enhanced accuracy of kinetic information from CT-CPMG experiments by transverse rotating-frame spectroscopy. Journal of Biomolecular NMR, 57(1), 73-82. doi:10.1007/s10858-013-9769-.


Cite as: http://hdl.handle.net/11858/00-001M-0000-0014-5574-F
Abstract
Micro-to-millisecond motions of proteins transmit pivotal signals for protein function. A powerful technique for the measurement of these motions is nuclear magnetic resonance spectroscopy. One of the most widely used methodologies for this purpose is the constant-time Carr-Purcell-Meiboom-Gill (CT-CPMG) relaxation dispersion experiment where kinetic and structural information can be obtained at atomic resolution. Extraction of accurate kinetics determined from CT-CPMG data requires refocusing frequencies that are much larger than the nuclei's exchange rate between states. We investigated the effect when fast processes are probed by CT-CPMG experiments via simulation and show that if the intrinsic relaxation rate is not known a priori the extraction of accurate kinetics is hindered. Errors on the order of 50 % in the exchange rate are attained when processes become fast, but are minimized to 5 % with a priori information. To alleviate this shortcoming, we developed an experimental scheme probing with large amplitude spin-lock fields, which specifically contains the intrinsic proton longitudinal Eigenrelaxation rate. Our approach was validated with ubiquitin and the Oscillatoria agardhii agglutinin (OAA). For OAA, an underestimation of 66 % in the kinetic rates was observed if is not included during the analysis of CT-CPMG data and result in incorrect kinetics and imprecise amplitude information. This was overcome by combining CT-CPMG with measured with a high power R-1 rho experiment. In addition, the measurement of removes the ambiguities in choosing between different models that describe CT-CPMG data.