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

• Published in: Nature (London) Vol. 420; no. 6911; pp. 98 - 102
• Main Authors: Oschkinat, Hartmut, Castellani, Federica, van Rossum, Barth, Diehl, Annette, Schubert, Mario, Rehbein, Kristina
• Format: Journal Article
• Language: English
• Published: London Nature Publishing 07.11.2002; Nature Publishing Group
• Subjects: Analytical chemistry; Animals; Biological and medical sciences; Brain; Carbon sources; Chemistry; Chickens; Crystals; Diffusion; Exact sciences and technology; Fundamental and applied biological sciences. Psychology; Glycerol; Magnetic Resonance Spectroscopy - methods; Membranes; Models, Molecular; Molecular biophysics; NMR; Nuclear magnetic resonance; Protein Conformation; Proteins; Protons; Resonance; Spectrin - chemistry; Spectrometric and optical methods; Spin Labels; src Homology Domains; Structure in molecular biology; Tridimensional structure; Magic angle; NMR spectrometer; Solid state; Spinning particle; Structure; Protein
• ISSN: 0028-0836; 1476-4687
• DOI: 10.1038/nature01070

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 systems or membrane proteins. 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) domain 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.