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CPMG Relaxation Dispersion

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Protein Dynamics

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1084))

Abstract

NMR relaxation is sensitive to molecular and internal motion of proteins. 15N longitudinal relaxation rate (R 1), transverse relaxation rate (R 2), and {1H}-15N Nuclear Overhauser Effect (NOE) experiments are often performed to globally elucidate protein dynamics, primarily on the sub-nanosecond timescale. In contrast, constant relaxation time R 2 dispersion experiments are applied to characterize protein equilibrium conformations that interconvert on the millisecond timescale. Information on local conformational equilibria of proteins provides important insights about protein energy landscapes and is useful to interpret molecular recognition mechanisms as well. Here, we describe a protocol for performing 15N Carr–Purcell–Meiboom–Gill (CPMG) R 2 dispersion measurements in solution, including protein preparation, step-by-step experimental parameter settings, and the first step of data analysis.

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Acknowledgments

The authors would like to thank Stefan Bagby and Dennis A. Torchia for critical reading of the manuscript. This project was supported by University of Pittsburgh.

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Authors and Affiliations

  1. Department of Structural Biology, University of Pittsburgh, Pittsburgh, PA, USA

    Rieko Ishima

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  1. Rieko Ishima
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Editors and Affiliations

  1. Department of Bioinformatics and Genomic, University of North Carolina at Charlott, Charlotte, North Carolina, USA

    Dennis R. Livesay

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Ishima, R. (2014). CPMG Relaxation Dispersion. In: Livesay, D. (eds) Protein Dynamics. Methods in Molecular Biology, vol 1084. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-658-0_2

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  • DOI: https://doi.org/10.1007/978-1-62703-658-0_2

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-657-3

  • Online ISBN: 978-1-62703-658-0

  • eBook Packages: Springer Protocols

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