Related Articles(13)C (?) decoupling during direct observation of carbonyl resonances in solution NMR of isotopically enriched proteins.
J Biomol NMR. 2014 Aug 17;
Authors: Ying J, Li F, Lee JH, Bax A
Abstract
Direct detection of (13)C can be advantageous when studying uniformly enriched proteins, in particular for paramagnetic proteins or when hydrogen exchange with solvent is fast. A scheme recently introduced for long-observation-window band-selective homonuclear decoupling in solid state NMR, LOW-BASHD (Struppe et al. in J Magn Reson 236:89-94, 2013) is shown to be effective for (13)C(?) decoupling during direct (13)C' observation in solution NMR experiments too. For this purpose, adjustment of the decoupling pulse parameters and delays is demonstrated to be important for increasing spectral resolution, to reduce three-spin effects, and to decrease the intensity of decoupling side-bands. LOW-BASHD then yields (13)C' line widths comparable to those obtained with the popular IPAP method, while enhancing sensitivity by ca 35*%. As a practical application of LOW-BASHD decoupling, requiring quantitative intensity measurement over a wide dynamic range, the impact of lipid binding on the (13)C'-detected NCO spectrum of the intrinsically disordered protein ?-synuclein is compared with that on the (1)H-detected (1)H-(15)N HSQC spectrum. Results confirm that synuclein's "dark state" behavior is not caused by paramagnetic relaxation or rapid hydrogen exchange.
PMID: 25129622 [PubMed - as supplied by publisher]
13Cα decoupling during direct observation of carbonyl resonances in solution NMR of isotopically enriched proteins
13Cα decoupling during direct observation of carbonyl resonances in solution NMR of isotopically enriched proteins
Abstract
Direct detection of 13C can be advantageous when studying uniformly enriched proteins, in particular for paramagnetic proteins or when hydrogen exchange with solvent is fast. A scheme recently introduced for long-observation-window band-selective homonuclear decoupling in solid state NMR, LOW-BASHD (Struppe et al. in J Magn Reson 236:89â??94, 2013) is shown to be effective for 13Cα decoupling during direct 13Câ?² observation in...
[NMR paper] Long-Observation-Window Band-Selective Homonuclear Decoupling: Increased Sensitivity and Resolution in Solid-State NMR Spectroscopy of Proteins
Long-Observation-Window Band-Selective Homonuclear Decoupling: Increased Sensitivity and Resolution in Solid-State NMR Spectroscopy of Proteins
Publication date: Available online 13 September 2013
Source:Journal of Magnetic Resonance</br>
Author(s): Jochem O. Struppe , Chen Yang , Yachong Wang , Roy V. Hernandez , Lisa M. Shamansky , Leonard J. Mueller</br>
Sensitivity and resolution are the two fundamental obstacles to extending solid-state nuclear magnetic resonance to even larger protein systems. Here, a novel long-observation-window band-selective...
nmrlearner
Journal club
0
09-13-2013 12:05 PM
Three-dimensional triple-resonance NMR Spectroscopy of isotopically enriched proteins
Three-dimensional triple-resonance NMR Spectroscopy of isotopically enriched proteins
Publication year: 2011
Source: Journal of Magnetic Resonance, Volume 213, Issue 2, December 2011, Pages 423-441</br>
Lewis E.*Kay, Mitsuhiko*Ikura, Rolf*Tschudin, Ad*Bax</br>
Four new and complementary three-dimensional triple-resonance experiments are described for obtaining complete backboneH,C, andN resonance assignments of proteins uniformly enriched withC andN. The new methods all rely onH detection and use multiple magnetization transfers through well-resolved one-bondJcouplings. Therefore, the...
nmrlearner
Journal club
0
12-11-2011 07:57 AM
Mammalian production of an isotopically enriched outer domain of the HIV-1 gp120 glycoprotein for NMR spectroscopy
Mammalian production of an isotopically enriched outer domain of the HIV-1 gp120 glycoprotein for NMR spectroscopy
Abstract NMR spectroscopic characterization of the structure or the dynamics of proteins generally requires the production of samples isotopically enriched in 15N, 13C, or 2H. The bacterial expression systems currently in use to obtain isotopic enrichment, however, cannot produce a number of eukaryotic proteins, especially those that require post-translational modifications such as N-linked glycosylation for proper folding or activity. Here, we report the use of an...
nmrlearner
Journal club
0
06-15-2011 02:31 AM
Direct observation of minimum-sized amyloid fibrils using solution NMR spectroscopy.
Direct observation of minimum-sized amyloid fibrils using solution NMR spectroscopy.
Related Articles Direct observation of minimum-sized amyloid fibrils using solution NMR spectroscopy.
Protein Sci. 2010 Oct 8;
Authors: Yoshimura Y, Sakurai K, Lee YH, Ikegami T, Chatani E, Naiki H, Goto Y
It is challenging to investigate the structure and dynamics of amyloid fibrils at the residue and atomic resolution due to their high molecular weight and heterogeneous properties. Here, we employed solution nuclear magnetic resonance (NMR) spectroscopy to...
nmrlearner
Journal club
0
10-12-2010 02:52 PM
[NMR paper] Multidimensional triple resonance NMR spectroscopy of isotopically uniformly enriched
Multidimensional triple resonance NMR spectroscopy of isotopically uniformly enriched proteins: a powerful new strategy for structure determination.
Related Articles Multidimensional triple resonance NMR spectroscopy of isotopically uniformly enriched proteins: a powerful new strategy for structure determination.
Ciba Found Symp. 1991;161:108-19; discussion 119-35
Authors: Bax A, Ikura M, Kay LE, Barbato G, Spera S
A procedure is described that affords complete 1H, 13C and 15N resonance assignment in proteins of up to about 25 kDa. The new...
nmrlearner
Journal club
0
08-21-2010 11:16 PM
Narrow carbonyl resonances in proton-diluted proteins facilitate NMR assignments in t
Abstract HNCO/HNCACO type correlation experiments are an alternative for assignment of backbone resonances in extensively deuterated proteins in the solid-state, given the fact that line widths on the order of 14â??17 Hz are achieved in the carbonyl dimension without the need of high power decoupling. The achieved resolution demonstrates that MAS solid-state NMR on extensively deuterated proteins is able to compete with solution-state NMR spectroscopy if proteins are investigated with correlation times Ï? c that exceed 25 ns.
Content Type Journal Article
DOI...