Abstract
Solid-state nuclear magnetic resonance (SSNMR) has become a prominent method in biology and is suitable for the characterization of insoluble proteins and protein aggregates such as amyloid fibrils, membrane–lipid complexes, and precipitated proteins. Often, the initial and the most critical step is to obtain spectroscopic assignments, that is, to determine chemical shifts of individual atoms. The procedures for SSNMR spectroscopic assignments are now well established for small microcrystalline proteins, where high signal-to-noise can be obtained. The sensitivity of the experiments and spectral resolution decrease with the increasing molecular weight, which makes setting SSNMR experiments in large proteins a much more challenging and demanding procedure. Here, we describe the protocol for the most common set of 3D magic angle spinning (MAS) SSNMR experiments. While the procedures described in the text are well known to SSNMR practitioners, we hope they will be of interest to scientists interested in extending their repertoire of biophysical techniques.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
McDermott A, Polenova T (2007) Solid state NMR: new tools for insight into enzyme function. Curr Opin Struct Biol 17:617–622
Ramamoorthy A (2009) Beyond NMR spectra of antimicrobial peptides: dynamical images at atomic resolution and functional insights. Solid State Nucl Magn Reson 35:201–207
Renault M, Cukkemane A, Baldus M (2010) Solid-state NMR spectroscopy on complex biomolecules. Angew Chem Int Ed Engl 49:8346–8357
Rienstra CM, Tucker-Kellogg L, Jaroniec CP, Hohwy M, Reif B, McMahon MT, Tidor B, Lozano-Perez T, Griffin RG (2002) De novo determination of peptide structure with solid-state magic-angle spinning NMR spectroscopy. Proc Natl Acad Sci USA 99:10260–10265
Castellani F, van Rossum B, Diehl A, Schubert M, Rehbein K, Oschkinat H (2002) Structure of a protein determined by solid-state magic-angle-spinning NMR spectroscopy. Nature 420:98–102
Pauli J, Baldus M, van Rossum B, de Groot H, Oschkinat H (2001) Backbone and side-chain C-13 and N-15 signal assignments of the alpha-spectrin SH3 domain by magic angle spinning solid-state NMR at 17.6 tesla. Chembiochem 2:272–281
Igumenova TI, McDermott AE, Zilm KW, Martin RW, Paulson EK, Wand AJ (2004) Assignments of carbon NMR resonances for microcrystalline ubiquitin. J Am Chem Soc 126:6720–6727
Bockmann A, Lange A, Galinier A, Luca S, Giraud N, Juy M, Heise H, Montserret R, Penin F, Baldus M (2003) Solid state NMR sequential resonance assignments and conformational analysis of the 2 × 10.4 kDa dimeric form of the Bacillus subtilis protein Crh. J Biomol NMR 27:323–339
Li Y, Berthold DA, Gennis RB, Rienstra CM (2008) Chemical shift assignment of the transmembrane helices of DsbB, a 20-kDa integral membrane enzyme, by 3D magic-angle spinning NMR spectroscopy. Protein Sci 17:199–204
Schuetz A, Wasmer C, Habenstein B, Verel R, Greenwald J, Riek R, Bockmann A, Meier BH (2010) Protocols for the sequential solid-state NMR spectroscopic assignment of a uniformly labeled 25 kDa protein: HET-s(1-227). Chembiochem 11:1543–1551
Sperling LJ, Berthold DA, Sasser TL, Jeisy-Scott V, Rienstra CM (2010) Assignment strategies for large proteins by magic-angle spinning NMR: the 21-kDa disulfide-bond-forming enzyme DsbA. J Mol Biol 399:268–282
Helmus JJ, Surewicz K, Nadaud PS, Surewicz WK, Jaroniec CP (2008) Molecular conformation and dynamics of the Y145Stop variant of human prion protein. Proc Natl Acad Sci USA 105:6284–6289
Shi L, Kawamura I, Jung KH, Brown LS, Ladizhansky V (2011) Conformation of a seven-helical transmembrane photosensor in the lipid environment. Angew Chem Int Ed Engl 50(6):1302–1305
Tycko R, Hu KN (2010) A Monte Carlo/simulated annealing algorithm for sequential resonance assignment in solid state NMR of uniformly labeled proteins with magic-angle spinning. J Magn Reson 205:304–314
Moseley HN, Sperling LJ, Rienstra CM (2010) Automated protein resonance assignments of magic angle spinning solid-state NMR spectra of beta1 immunoglobulin binding domain of protein G (GB1). J Biomol NMR 48:123–128
Shi L, Ahmed MAM, Zhang W, Whited G, Brown LS, Ladizhansky V (2009) Three-dimensional solid-state NMR study of a seven-helical integral membrane proton pump—structural insights. J Mol Biol 386:1078–1093
Shi L, Lake EM, Ahmed MA, Brown LS, Ladizhansky V (2009) Solid-state NMR study of proteorhodopsin in the lipid environment: secondary structure and dynamics. Biochim Biophys Acta 1788:2563–2574
Sperling LJ, Nieuwkoop AJ, Lipton AS, Berthold DA, Rienstra CM (2010) High resolution NMR spectroscopy of nanocrystalline proteins at ultra-high magnetic field. J Biomol NMR 46:149–155
Pines A, Gibby MG, Waugh JS (1973) Proton-enhanced NMR of dilute spins in solids. J Chem Phys 59:569–590
Morcombe CR, Zilm KW (2003) Chemical shift referencing in MAS solid state NMR. J Magn Reson 162:479–486
Andrew ER, Bradbury A, Eades RG, Wynn VT (1966) Resonant rotational broadening of nuclear magnetic resonance spectra. Phys Lett 21:505–506
Raleigh DP, Levitt MH, Griffin RG (1988) Rotational resonance in solid-state NMR. Chem Phys Lett 146:71–76
Fung BM, Khitrin AK, Ermolaev K (2000) An improved broadband decoupling sequence for liquid crystals and solids. J Magn Reson 142:97–101
Baldus M, Petkova AT, Herzfeld J, Griffin RG (1998) Cross polarization in the tilted frame: assignment and spectral simplification in heteronuclear spin systems. Mol Phys 95:1197–1207
Takegoshi K, Nakamura S, Terao T (2001) C-13-H-1 dipolar-assisted rotational resonance in magic-angle spinning NMR. Chem Phys Lett 344:631–637
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2012 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Shi, L., Ladizhansky, V. (2012). Magic Angle Spinning Solid-State NMR Experiments for Structural Characterization of Proteins. In: Uversky, V., Dunker, A. (eds) Intrinsically Disordered Protein Analysis. Methods in Molecular Biology, vol 895. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-927-3_12
Download citation
DOI: https://doi.org/10.1007/978-1-61779-927-3_12
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-61779-926-6
Online ISBN: 978-1-61779-927-3
eBook Packages: Springer Protocols