NMR spectroscopic assignment of backbone and side-chain protons in fully protonated proteins: microcrystals, sedimented assemblies, and amyloid fibrils
Authors
Jan Stanek, Universite de Lyon
Loren Andreas, Universite de Lyon
Kristaps Jaudzems, Institute of Organic Synthesis, Universite de Lyon
Diane Cala, Universite de Lyon
Daniela Lalli, Universite de Lyon
Andrea Bertarello, Universite de Lyon
Tobias Schubeis, Universite de Lyon
Inara Akopjana, Biomedical Research and Study Centre
Svetlana Kotelovica, Biomedical Research and Study Centre
Kaspars Tars, Biomedical Research and Study Centre
Andrea Pica, University of Naples
Serena Leone, University of Naples
D Picone, University of Naples
Zhi-Qiang Xu, University of WollongongFollow
Nicholas E. Dixon, University of WollongongFollow
Denis Martinez, University of Bordeaux
Melanie Berbon, University of Bordeaux
Nadia El Mammeri, University of Bordeaux
Abdelmajid Noubhani, University of Bordeaux
Sven Saupe, University of Bordeaux
Birgit Habenstein, University of Bordeaux
Antoine Loquet, University of Bordeaux
Guido Pintacuda, University of LyonFollow
Abstract
We demonstrate sensitive detection of alpha protons of fully protonated proteins by solid-state NMR spectroscopy with 100-111 kHz magic-angle spinning (MAS). The excellent resolution in the Cα-Hα plane is demonstrated for 5 proteins, including microcrystals, a sedimented complex, a capsid and amyloid fibrils. A set of 3D spectra based on a Cα-Hα detection block was developed and applied for the sequence-specific backbone and aliphatic side-chain resonance assignment using only 500 μg of sample. These developments accelerate structural studies of biomolecular assemblies available in submilligram quantities without the need of protein deuteration.
Publication Details
Stanek, J., Andreas, L. B., Jaudzems, K., Cala, D., Lalli, D., Bertarello, A., Schubeis, T., Akopjana, I., Kotelovica, S., Tars, K., Pica, A., Leone, S., Picone, D., Xu, Z., Dixon, N. E., Martinez, D., Berbon, M., El Mammeri, N., Noubhani, A., Saupe, S., Habenstein, B., Loquet, A. & Pintacuda, G. (2016). NMR spectroscopic assignment of backbone and side-chain protons in fully protonated proteins: microcrystals, sedimented assemblies, and amyloid fibrils. Angewandte Chemie International Edition, 55 (50), 15504-15509.