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Structure of a protein determined by solid-state magic-angle-spinning NMR spectroscopy

Abstract

The determination of a representative set of protein structures is a chief aim in structural genomics. Solid-state NMR may have a crucial role in structural investigations of those proteins that do not easily form crystals or are not accessible to solution NMR, such as amyloid systems1 or membrane proteins2,3,4. Here we present a protein structure determined by solid-state magic-angle-spinning (MAS) NMR. Almost complete 13C and 15N resonance assignments for a micro-crystalline preparation of the α-spectrin Src-homology 3 (SH3) domain5 formed the basis for the extraction of a set of distance restraints. These restraints were derived from proton-driven spin diffusion (PDSD) spectra of biosynthetically site-directed, labelled samples obtained from bacteria grown using [1,3-13C]glycerol or [2-13C]glycerol as carbon sources. This allowed the observation of long-range distance correlations up to 7 Å. The calculated global fold of the α-spectrin SH3 domain is based on 286 inter-residue 13C–13C and six 15N–15N restraints, all self-consistently obtained by solid-state MAS NMR. This MAS NMR procedure should be widely applicable to small membrane proteins that can be expressed in bacteria.

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Figure 1: Labelling patterns and NMR spectra for the different α-spectrin SH3 domain preparations.
Figure 2: Assignment strategy.
Figure 3: Solid-state structure of the α-spectrin SH3 domain.

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Acknowledgements

H. de Groot is acknowledged for access to the high-field NMR facility in Leiden. The authors thank R. Kühne, P. Schmieder, G. Krause and C. Glaubitz for discussions, and L. Ball, K. Heuer and K. Zierler for carefully reading the manuscript.

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Correspondence to Hartmut Oschkinat.

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Castellani, F., van Rossum, B., Diehl, A. et al. Structure of a protein determined by solid-state magic-angle-spinning NMR spectroscopy. Nature 420, 99–102 (2002). https://doi.org/10.1038/nature01070

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