Rapid Proton-Detected NMR Assignment for Proteins with Fast Magic Angle Spinning
Emeline Barbet-Massin
(1)
,
Andrew J. Pell
(2)
,
Joren S. Retel
(3)
,
Loren B Andreas
(2, 4, 5)
,
Kristaps Jaudzems
(6)
,
W. Trent Franks
(3)
,
Andrew J Nieuwkoop
(3)
,
Matthias Hiller
(3)
,
Victoria Higman
(3)
,
Paul Guerry
(2)
,
Andrea Bertarello
(2)
,
Michael J. Knight
(2)
,
Michèle Felletti
(2)
,
Tanguy Le Marchand
(2)
,
Svetlana Kotelovica
(7)
,
Inara Akopjana
(7)
,
Kaspars Tars
(7)
,
M. Stoppini
(8)
,
Vittorio Bellotti
(8, 9)
,
Martino Bolognesi
(10)
,
Stefano Ricagno
(10)
,
James J Chou
(11)
,
Robert G Griffin
(4, 5)
,
Hartmut Oschkinat
(3)
,
Anne Lesage
(12)
,
Lyndon Emsley
(12)
,
Torsten Herrmann
,
Guido Pintacuda
(1)
1
Biological Solid-State NMR Methods - Méthodes de RMN à l'état solide en biologie
2 ISA - Institut des Sciences Analytiques
3 FMP - Leibniz Forschungsinstitut für Molekulare Pharmakolgie = Leibniz Institute for Molecular Pharmacology [Berlin, Allemagne]
4 MIT, Dept Chem
5 MIT - Francis Bitter Magnet Lab
6 Latvian Institute of Organic Synthesis [Riga, Lituania]
7 Biomedical Research & Study Centre
8 Dept Mol Med
9 UCL, Ctr Amyloidosis & Acute Phase Prot
10 Department of Biomolecular Sciences and Biotechnology
11 School of Medicine
12 Solid-State NMR Methods for Materials - Méthodes de RMN à l'état solide pour les matériaux
2 ISA - Institut des Sciences Analytiques
3 FMP - Leibniz Forschungsinstitut für Molekulare Pharmakolgie = Leibniz Institute for Molecular Pharmacology [Berlin, Allemagne]
4 MIT, Dept Chem
5 MIT - Francis Bitter Magnet Lab
6 Latvian Institute of Organic Synthesis [Riga, Lituania]
7 Biomedical Research & Study Centre
8 Dept Mol Med
9 UCL, Ctr Amyloidosis & Acute Phase Prot
10 Department of Biomolecular Sciences and Biotechnology
11 School of Medicine
12 Solid-State NMR Methods for Materials - Méthodes de RMN à l'état solide pour les matériaux
Andrea Bertarello
- Fonction : Auteur
- PersonId : 772433
- ORCID : 0000-0003-3705-1760
Martino Bolognesi
- Fonction : Auteur
- PersonId : 908158
Hartmut Oschkinat
- Fonction : Auteur
- PersonId : 925444
- ORCID : 0000-0002-4384-9544
Anne Lesage
- Fonction : Auteur
- PersonId : 960699
Lyndon Emsley
- Fonction : Auteur
- PersonId : 960691
Torsten Herrmann
- Fonction : Auteur
- PersonId : 742391
- IdHAL : torsten-herrmann
- ORCID : 0000-0003-2115-4781
Guido Pintacuda
- Fonction : Auteur
- PersonId : 960700
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.