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

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-.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-0014-5574-F Version Permalink: http://hdl.handle.net/11858/00-001M-0000-0028-DD43-3
Genre: Journal Article

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 Creators:
Ban, D.1, Author              
Mazur, A.1, Author              
Giao Carneiro, M.1, Author              
Sabo, T. M.1, Author              
Giller, K.1, Author              
Koharudin, L. M. I., Author
Becker, S.1, Author              
Gronenborn, A. M., Author
Griesinger, C.1, Author              
Lee, D.1, Author              
Affiliations:
1Department of NMR-based Structural Biology, MPI for biophysical chemistry, Max Planck Society, ou_578567              

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Free keywords: Relaxation dispersion; CT-CPMG; R-1 rho; Kinetics
 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.

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Language(s): eng - English
 Dates: 2013-08-152013-09
 Publication Status: Published in print
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 Rev. Method: Peer
 Identifiers: DOI: 10.1007/s10858-013-9769-
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Title: Journal of Biomolecular NMR
Source Genre: Journal
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Pages: - Volume / Issue: 57 (1) Sequence Number: - Start / End Page: 73 - 82 Identifier: -