Data di Pubblicazione:
2014
Citazione:
Rapid proton-detected NMR assignment for proteins with fast magic angle spinning / E. Barbet-Massin, A.J. Pell, J.S. Retel, L.B. Andreas, K. Jaudzems, W..T. Franks, A.J. Nieuwkoop, M. Hiller, V. Higman, P. Guerry, A. Bertarello, M.J. Knight, M. Felletti, T. Le Marchand, S. Kotelovica, I. Akopjana, K. Tars, M. Stoppini, V. Bellotti, M. Bolognesi, S. Ricagno, J.J. Chou, R.G. Griffin, H. Oschkinat, A. Lesage, L. Emsley, T. Herrmann, G. Pintacuda. - In: JOURNAL OF THE AMERICAN CHEMICAL SOCIETY. - ISSN 0002-7863. - 136:35(2014 Sep 03), pp. 12489-12497. [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.
Tipologia IRIS:
01 - Articolo su periodico
Keywords:
Chemistry (all); Catalysis; Biochemistry; Colloid and Surface Chemistry
Elenco autori:
E. Barbet Massin, A.J. Pell, J.S. Retel, L.B. Andreas, K. Jaudzems, W..T. Franks, A.J. Nieuwkoop, M. Hiller, V. Higman, P. Guerry, A. Bertarello, M.J. Knight, M. Felletti, T. Le Marchand, S. Kotelovica, I. Akopjana, K. Tars, M. Stoppini, V. Bellotti, M. Bolognesi, S. Ricagno, J.J. Chou, R.G. Griffin, H. Oschkinat, A. Lesage, L. Emsley, T. Herrmann, G. Pintacuda
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