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Solid State Nucl Magn Reson. 2017 Oct;87:86-95. doi: 10.1016/j.ssnmr.2017.04.002. Epub 2017 Apr 14.

Protein conformational dynamics studied by 15N and 1H R relaxation dispersion: Application to wild-type and G53A ubiquitin crystals.

Author information

1
Université Grenoble Alpes, IBS, F-38044 Grenoble, France; CEA, Institut de Biologie Structurale, F-38044 Grenoble, France; CNRS, Institut de Biologie Structurale, F-38044 Grenoble, France.
2
Ludwig-Maximilians-Universität, Department Chemie, D-81377 München, Germany.
3
Université Grenoble Alpes, IBS, F-38044 Grenoble, France; CEA, Institut de Biologie Structurale, F-38044 Grenoble, France; CNRS, Institut de Biologie Structurale, F-38044 Grenoble, France. Electronic address: paul.schanda@ibs.fr.

Abstract

Solid-state NMR spectroscopy can provide site-resolved information about protein dynamics over many time scales. Here we combine protein deuteration, fast magic-angle spinning (~45-60kHz) and proton detection to study dynamics of ubiquitin in microcrystals, and in particular a mutant in a region that undergoes microsecond motions in a β-turn region in the wild-type protein. We use 15N R relaxation measurements as a function of the radio-frequency (RF) field strength, i.e. relaxation dispersion, to probe how the G53A mutation alters these dynamics. We report a population-inversion of conformational states: the conformation that in the wild-type protein is populated only sparsely becomes the predominant state. We furthermore explore the potential to use amide-1H R relaxation to obtain insight into dynamics. We show that while quantitative interpretation of 1H relaxation remains beyond reach under the experimental conditions, due to coherent contributions to decay, one may extract qualitative information about flexibility.

KEYWORDS:

Fast MAS; Protein dynamics; Proton detection; Proton relaxation; Solid-state NMR; Spin relaxation; β-turn

PMID:
28438365
PMCID:
PMC5531261
DOI:
10.1016/j.ssnmr.2017.04.002
[Indexed for MEDLINE]
Free PMC Article

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