Rapid proton-detected NMR assignment for proteins with fast magic angle spinning

Emeline Barbet-Massin, Andrew J. Pell, Joren S. Retel, Loren B. Andreas, Kristaps Jaudzems, W. Trent Franks, Andrew J. Nieuwkoop, Matthias Hiller, Victoria Higman, Paul Guerry, Andrea Bertarello, Michael J. Knight, Michele Felletti, Tanguy Le Marchand, Svetlana Kotelovica, Inara Akopjana, Kaspars Tars, Monica Stoppini, Vittorio Bellotti, Martino BolognesiStefano Ricagno, James J. Chou, Robert G. Griffin, Hartmut Oschkinat, Anne Lesage, Lyndon Emsley, Torsten Herrmann, Guido Pintacuda*

*Corresponding author for this work

Research output: Contribution to journalArticle (Academic Journal)peer-review

165 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)12489-12497
Number of pages9
JournalJournal of the American Chemical Society
Volume136
Issue number35
DOIs
Publication statusPublished - 3 Sep 2014

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