Centre de RMN à Très Hauts Champs, CNRS/Université Claude Bernard Lyon 1/Ecole Normale Supérieure de Lyon, Villeurbanne, France
Fast magic-angle spinning NMR of membrane proteins
Building on a decade of continuous advances of the community, the recent development of very fast (60 kHz and above) magic-angle spinning (MAS) probes has revolutionised the field of solid-state NMR. Today, rapid “fingerprinting” of proteins by 1H detection is possible with a ten-fold reduction of the required sample amounts with respect to conventional approaches, not only in deuterated molecules but also in fully-protonated substrates. Extensive and robust resonance assignments can be derived rapidly for small-to-medium sized proteins (up to ca. 300 residues), opening the way to the determination of inter-nuclear proximities, relative orientations of secondary structural elements, protein-cofactor interactions, local and global dynamics.
Fast MAS and 1H detection techniques have nowadays been shown to be applicable to membrane-bound systems. This talk reviews the strategies underlying this recent leap forward in sensitivity and resolution, describing its potential for the detailed characterization of membrane proteins in lipid bilayers.
- T. Schubeis, T. Le Marchand, C. Daday, W. Kopec, K.T. Movellan, J. Stanek, T.S. Schwarzer, K Castiglione, B.L. de Groot, G. Pintacuda, and L. B. Andreas (2020) A β-Barrel for Oil Transport through Lipid Membranes: Dynamic NMR Structures of AlkL Proc Natl Acad Sci USA 117, 21014–21
- J. Stanek, T. Schubeis, P. Paluch., P. Güntert, L. Andreas, and G. Pintacuda (2020) Automated backbone NMR resonance assignment of large proteins using redundant linking from a single simultaneous acquisition J Am Chem Soc 142, 5793-5799
- T. Schubeis, T. Le Marchand, L. B. Andreas, G. Pintacuda (2018) 1H magic-angle spinning NMR evolves as a powerful new tool for membrane proteins J Magn Reson 287, 140-152
- J. S. Retel, A. J. Nieuwkoop, M. Hiller, V. A. Higman, E. Barbet-Massin, J. Stanek, L. B. Andreas, W. T. Franks, B. J. van Rossum, K. R. Vinothkumar, L. Handel, G. G. de Palma, B. Bardiaux, G. Pintacuda, L. Emsley, W. Kuhlbrandt, & H. Oschkinat (2017) Structure of outer membrane protein G in lipid bilayers Nat Commun 8, 2073
- D. Lalli, M. Idso, L. B. Andreas, S. Hussain, N. Baxter, S. Han, B. F. Chmelkaand G. Pintacuda (2017) Proton-based structural analysis of a heptahelical transmembrane protein in lipid bilayers J Am Chem Soc 139, 13006-13012
- O. Saurel, I. Iordanov, G. Nars, P. Demange, T. Le Marchand, L. B. Andreas, G. Pintacuda, A. Milon (2017) Local and global dynamics in Klebsiella pneumoniae outer membrane protein A in lipid bilayers probed at atomic resolution J Am Chem Soc 139, 1590-1597
- J. Stanek, L. B. Andreas, K. Jaudzems, D. Cala, D. Lalli, A. Bertarello, T. Schubeis, I. Akopjana, S. Kotelovica, K. Tars, A. Pica, S. Leone, D. Picone, Z.-Q. Xu, N. E. Dixon, D. Martinez, M. Berbon, N. El Mammeri, A. Noubhani, S. Saupe, B. Habenstein, A. Loquet, G. Pintacuda (2016) Backbone and side-chain proton NMR assignment in fully protonated proteins: microcrystals, sedimented assemblies, and amyloid fibrils Angew Chem Int Ed Engl 55, 15504-15509; Angew Chem 128, 15730-15735 (frontispice article)
- L. B. Andreas, K. Jaudzems, J. Stanek, D. Lalli, A. Bertarello, T. Le Marchand, D. Cala-De Paepe, S. Kotelovica, I. Akopjana, B. Knott, S. Wegner, F. Engelke, A. Lesage, L. Emsley, K. Tars, T. Herrmann, and G. Pintacuda (2016) Structure of fully protonated proteins by proton-detected magic-angle spinning NMR Proc Natl Acad Sci USA 113, 9187-9192
Contact: Olivier Saurel (Olivier.Saurel@ipbs.fr)
Link to attend the seminar: https://meet.starleaf.com/4376320706/app
11:00 - 12:00