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Article Dans Une Revue Journal of the American Chemical Society Année : 2014

Rapid Proton-Detected NMR Assignment for Proteins with Fast Magic Angle Spinning

Andrea Bertarello
M. Stoppini
  • Fonction : Auteur
James J Chou
  • Fonction : Auteur
Torsten Herrmann

Résumé

Using a set of six H-1-detected triple-resonance NMR experiments, we establish a method for sequence-specific backbone resonance assignment of magic angle spinning (MAS) nuclear magnetic resonance (NMR) spectra of 5-30 kDa proteins. The approach relies on perdeuteration, amide H-2/H-1 exchange, high magnetic fields, and high-spinning frequencies (omega(r)/2 pi >= 60 kHz) and yields high-quality NMR data, enabling the use of automated analysis. The method is validated with five examples of proteins in different condensed states, including two microcrystalline proteins, a sedimented virus capsid, and two membrane-embedded systems. In comparison to contemporary C-13/N-15-based methods, this approach facilitates and accelerates the MAS NMR assignment process, shortening the spectral acquisition times and enabling the use of unsupervised state-of-the-art computational data analysis protocols originally developed for solution NMR.

Dates et versions

hal-01070782 , version 1 (02-10-2014)

Identifiants

Citer

Emeline Barbet-Massin, Andrew J. Pell, Joren S. Retel, Loren B Andreas, Kristaps Jaudzems, et al.. Rapid Proton-Detected NMR Assignment for Proteins with Fast Magic Angle Spinning. Journal of the American Chemical Society, 2014, 136 (35), pp.12489-12497. ⟨10.1021/ja507382j⟩. ⟨hal-01070782⟩
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