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Determination of Protein ps-ns Motions by High-Resolution Relaxometry

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Protein NMR

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1688))

Abstract

Many of the functions of biomacromolecules can be rationalized by the characterization of their conformational energy landscapes: the structures of the dominant states, transitions between states and motions within states. Nuclear magnetic resonance (NMR) spectroscopy is the technique of choice to study internal motions in proteins. The determination of motions on picosecond to nanosecond timescales requires the measurement of nuclear spin relaxation rates at multiple magnetic fields. High sensitivity and resolution are obtained only at high magnetic fields, so that, until recently, site-specific relaxation rates in biomolecules were only measured over a narrow range of high magnetic fields. This limitation was particularly striking for the quantification of motions on nanosecond timescales, close to the correlation time for overall rotational diffusion. High-resolution relaxometry is an emerging technique to investigate picosecond—nanosecond motions of proteins. This approach uses a high-field NMR spectrometer equipped with a sample shuttle device, which allows for the measurement of the relaxation rate constants at low magnetic fields, while preserving the sensitivity and resolution of a high-field NMR spectrometer. The combined analysis of high-resolution relaxometry and standard high-field relaxation data provides a more accurate description of the dynamics of proteins, in particular in the nanosecond range. The purpose of this chapter is to describe how to perform high-resolution relaxometry experiments and how to analyze the rates measured with this technique.

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Acknowledgments

This work was funded by the European Research Council (ERC) under the European Community Seventh Framework Program (FP7/2007–2013), ERC Grant Agreement 279519 (2F4BIODYN).

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Author notes

  1. Samuel F. Cousin, Pavel Kadeřávek and Nicolas Bolik-Coulon contributed equally to this work.

Authors and Affiliations

  1. Laboratoire des Biomolécules, Département de Chimie, École Normale Supérieure - PSL Research University, UPMC Univ Paris 06, CNRS, 24 rue Lhomond, 75005, Paris, France

    Samuel F. Cousin, Pavel Kadeřávek, Nicolas Bolik-Coulon & Fabien Ferrage

  2. Sorbonne Universités, Ecole Normale Supérieure, CNRS, Laboratoire des Biomolécules (LBM), UPMC Univ Paris 06, Paris, France

    Samuel F. Cousin, Pavel Kadeřávek, Nicolas Bolik-Coulon & Fabien Ferrage

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  1. Samuel F. Cousin
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  2. Pavel Kadeřávek
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  3. Nicolas Bolik-Coulon
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  4. Fabien Ferrage
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Correspondence to Fabien Ferrage .

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Editors and Affiliations

  1. Department of Chemistry and Biochemistry, The City College of New York, New York, New York, USA

    Ranajeet Ghose

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Cousin, S.F., Kadeřávek, P., Bolik-Coulon, N., Ferrage, F. (2018). Determination of Protein ps-ns Motions by High-Resolution Relaxometry. In: Ghose, R. (eds) Protein NMR. Methods in Molecular Biology, vol 1688. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7386-6_9

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  • DOI: https://doi.org/10.1007/978-1-4939-7386-6_9

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