Rapid Proton-Detected NMR Assignment for Proteins with Fast Magic Angle Spinning
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Griffin_Rapid proton.pdf
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Author(s)
Barbet-Massin, Emeline
Pell, Andrew J.
Retel, Joren S.
Jaudzems, Kristaps
Franks, W. Trent
Nieuwkoop, Andrew J.
Hiller, Matthias
Higman, Victoria
Guerry, Paul
Bertarello, Andrea
Date Issued
August 2014
Journal
Journal of the American Chemical Society
Publisher
American Chemical Society (ACS)
Citation
Barbet-Massin, Emeline, Andrew J. Pell, Joren S. Retel, Loren B. Andreas, Kristaps Jaudzems, W. Trent Franks, Andrew J. Nieuwkoop, et al. “Rapid Proton-Detected NMR Assignment for Proteins with Fast Magic Angle Spinning.” Journal of the American Chemical Society 136, no. 35 (September 3, 2014): 12489-12497.
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Author's final manuscript
Abstract
Using a set of six [superscript 1]H-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 [superscript 2]H/[superscript 1]H exchange, high magnetic fields, and high-spinning frequencies (ω[subscript r]/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 [superscript 13]C/[superscript 15]N-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.
MIT Department
Massachusetts Institute of Technology. Department of Chemistry
Francis Bitter Magnet Laboratory (Massachusetts Institute of Technology)
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DOI of Published Version
https://doi.org/10.1021/ja507382j
