Rapid Proton-Detected NMR Assignment for Proteins with Fast Magic Angle Spinning
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Author
Barbet-Massin, Emeline
Pell, Andrew J.
Retel, Joren S.
Andreas, Loren B.
Jaudzems, Kristaps
Franks, W. Trent
Nieuwkoop, Andrew J.
Hiller, Matthias
Higman, Victoria
Guerry, Paul
Bertarello, Andrea
Knight, Michael J.
Felletti, Michele
Le Marchand, Tanguy
Kotelovica, Svetlana
Akopjana, Inara
Tars, Kaspars
Stoppini, Monica
Bellotti, Vittorio
Bolognesi, Martino
Ricagno, Stefano
Chou, James
J.
Griffin, Robert G.
Oschkinat, Hartmut
Lesage, Anne
Emsley, Lyndon
Herrmann, Torsten
Pintacuda, Guido
Note: Order does not necessarily reflect citation order of authors.
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https://doi.org/10.1021/ja507382jMetadata
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Barbet-Massin, E., A. J. Pell, J. S. Retel, L. B. Andreas, K. Jaudzems, W. T. Franks, A. J. Nieuwkoop, et al. 2014. “Rapid Proton-Detected NMR Assignment for Proteins with Fast Magic Angle Spinning.” Journal of the American Chemical Society 136 (35): 12489-12497. doi:10.1021/ja507382j. http://dx.doi.org/10.1021/ja507382j.Abstract
Using a set of six 1H-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 2H/1H exchange, high magnetic fields, and high-spinning frequencies (ωr/2π ≥ 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 13C/15N-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.Other Sources
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4156866/pdf/Terms of Use
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