Enhanced Resolution and Coherence Lifetimes in the Solid-State NMR Spectroscopy of Perdeuterated Proteins under Ultrafast Magic-Angle Spinning

We investigate the combined effect of perdeuteration and fast magic-angle spinning on the resolution and sensitivity of proton-detected protein NMR spectra and on coherence lifetimes. With 60 kHz spinning of a microcrystalline α-spectrin SH3 sample at a field strength of 23 T, a regime is attained where there is no substantial difference in resolution between perdeuterated samples with 10 or 100% protons at the exchangeable sites.1H resolution is then limited by inhomogeneous contributions. Upon fast spinning, the most dramatic line narrowing effects are observed for residues in the loop or bend regions of the protein, probably due to the removal of destructive dynamics effects. This investigation paves the way for using samples with 100% protons at the exchangeable sites in structure determination protocols, since all backbone amide sites can now contribute to the signal.

[1]  Ad Bax,et al.  Methodological advances in protein NMR , 1993 .

[2]  P. Schmieder,et al.  High resolution 1H-detected solid-state NMR spectroscopy of protein aliphatic resonances: access to tertiary structure information. , 2010, Journal of the American Chemical Society.

[3]  B. Reif,et al.  Ultrahigh resolution in proton solid-state NMR spectroscopy at high levels of deuteration. , 2006, Angewandte Chemie.

[4]  Martin Blackledge,et al.  Anisotropic collective motion contributes to nuclear spin relaxation in crystalline proteins. , 2010, Journal of the American Chemical Society.

[5]  P. Hodgkinson.  High-Resolution 1H NMR Spectroscopy of Solids , 2011 .

[6]  U. Fink,et al.  Proton-detected scalar coupling based assignment strategies in MAS solid-state NMR spectroscopy applied to perdeuterated proteins. , 2008, Journal of magnetic resonance.

[7]  D. Kern,et al.  Dynamic personalities of proteins , 2007, Nature.

[8]  Hartmut Oschkinat,et al.  Optimum levels of exchangeable protons in perdeuterated proteins for proton detection in MAS solid-state NMR spectroscopy , 2010, Journal of biomolecular NMR.

[9]  C. Rienstra,et al.  Proton-detected solid-state NMR spectroscopy of fully protonated proteins at 40 kHz magic-angle spinning. , 2007, Journal of the American Chemical Society.

[10]  Beat H. Meier,et al.  Amyloid Fibrils of the HET-s(218–289) Prion Form a β Solenoid with a Triangular Hydrophobic Core , 2008, Science.

[11]  Lyndon Emsley,et al.  Through-Bond Carbon−Carbon Connectivities in Disordered Solids by NMR , 1999 .

[12]  S. Grzesiek,et al.  Measurement of site-specific 13C spin-lattice relaxation in a crystalline protein. , 2010, Journal of the American Chemical Society.

[13]  I. Bertini,et al.  High-resolution and sensitivity through-bond correlations in ultra-fast magic angle spinning (MAS) solid-state NMR , 2011 .

[14]  A. McDermott.  Structure and dynamics of membrane proteins by magic angle spinning solid-state NMR. , 2009, Annual review of biophysics.

[15]  John S. Waugh,et al.  NMR in rotating solids , 1979 .

[16]  U. Fink,et al.  Assignment of dynamic regions in biological solids enabled by spin-state selective NMR experiments. , 2010, Journal of the American Chemical Society.

[17]  W. T. Franks,et al.  Sensitivity and resolution in proton solid-state NMR at intermediate deuteration levels: quantitative linewidth characterization and applications to correlation spectroscopy. , 2006, Journal of magnetic resonance.

[18]  E. Salager,et al.  Enhanced sensitivity in high-resolution 1H solid-state NMR spectroscopy with DUMBO dipolar decoupling under ultra-fast MAS , 2009 .

[19]  R G Griffin,et al.  1H-1H MAS correlation spectroscopy and distance measurements in a deuterated peptide. , 2001, Journal of magnetic resonance.

[20]  B. Reif,et al.  Quantitative measurement of differential 15NHα/βT2 relaxation rates in a perdeuterated protein by MAS solid‐state NMR spectroscopy , 2007 .

[21]  Roberta Pierattelli,et al.  Fast acquisition of multi-dimensional spectra in solid-state NMR enabled by ultra-fast MAS. , 2009, Journal of magnetic resonance.

[22]  B. Meier,et al.  Low-power XiX decoupling in MAS NMR experiments. , 2003, Journal of magnetic resonance.

[23]  B. Reif,et al.  Direct observation of millisecond to second motions in proteins by dipolar CODEX NMR spectroscopy. , 2009, Journal of the American Chemical Society.

[24]  Ronald Kühne,et al.  Solid-state NMR and SAXS studies provide a structural basis for the activation of αB-crystallin oligomers , 2010, Nature Structural &Molecular Biology.

[25]  B. Reif,et al.  TROSY effects in MAS solid-state NMR , 2008 .

[26]  B. Reif,et al.  1H detected 1H,15N correlation spectroscopy in rotating solids. , 2003, Journal of magnetic resonance.

[27]  Y. Ishii,et al.  Sensitivity enhancement in 13C solid-state NMR of protein microcrystals by use of paramagnetic metal ions for optimizing 1H T1 relaxation , 2007 .

[28]  R. Griffin,et al.  High-resolution solid-state NMR structure of a 17.6 kDa protein. , 2010, Journal of the American Chemical Society.

[29]  B. Reif,et al.  Deuterated peptides and proteins in MAS solid-state NMR , 2006 .

[30]  M. Schubert,et al.  Structure of a protein determined by solid-state magic-angle-spinning NMR spectroscopy , 2002, Nature.

[31]  Uwe Fink,et al.  Structure calculation from unambiguous long-range amide and methyl 1H-1H distance restraints for a microcrystalline protein with MAS solid-state NMR spectroscopy. , 2011, Journal of the American Chemical Society.

[32]  P. Schanda,et al.  Quantitative analysis of protein backbone dynamics in microcrystalline ubiquitin by solid-state NMR spectroscopy. , 2010, Journal of the American Chemical Society.

[33]  C. Rienstra,et al.  High resolution 13C-detected solid-state NMR spectroscopy of a deuterated protein , 2010, Journal of biomolecular NMR.

[34]  R. Tycko.  Solid-state NMR studies of amyloid fibril structure. , 2011, Annual review of physical chemistry.

[35]  U. Fink,et al.  Quantitative analysis of backbone motion in proteins using MAS solid-state NMR spectroscopy , 2009, Journal of biomolecular NMR.

[36]  Robert G Griffin,et al.  Proton assisted recoupling at high spinning frequencies. , 2009, The journal of physical chemistry. B.

[37]  I. Bertini,et al.  Solid-state NMR spectroscopy of a paramagnetic protein: assignment and study of human dimeric oxidized CuII-ZnII superoxide dismutase (SOD). , 2007, Angewandte Chemie.

[38]  Y. Ishii,et al.  Enhanced sensitivity and resolution in (1)H solid-state NMR spectroscopy of paramagnetic complexes under very fast magic angle spinning. , 2005, Journal of the American Chemical Society.

[39]  A. Lesage.  Recent advances in solid-state NMR spectroscopy of spin I=1/2 nuclei. , 2009, Physical chemistry chemical physics : PCCP.

[40]  Kurt W Zilm,et al.  Sensitive high resolution inverse detection NMR spectroscopy of proteins in the solid state. , 2003, Journal of the American Chemical Society.

[41]  M. Blackledge,et al.  Quantitative analysis of backbone dynamics in a crystalline protein from nitrogen-15 spin-lattice relaxation. , 2005, Journal of the American Chemical Society.

[42]  P. Schanda,et al.  Direct detection of (3h)J(NC') hydrogen-bond scalar couplings in proteins by solid-state NMR spectroscopy. , 2009, Angewandte Chemie.

[43]  Matthias Huber,et al.  A proton-detected 4D solid-state NMR experiment for protein structure determination. , 2011, Chemphyschem : a European journal of chemical physics and physical chemistry.

[44]  B. Reif,et al.  Microsecond time scale mobility in a solid protein as studied by the 15N R(1rho) site-specific NMR relaxation rates. , 2010, Journal of the American Chemical Society.

[45]  O. Pongs,et al.  A structural link between inactivation and block of a K+ channel , 2008, Nature Structural &Molecular Biology.