Direct detection of the TROSY component of proton-attached 15N nuclei (15N-detected TROSY) yields high quality spectra with high field magnets, by taking advantage of the slow 15N transverse relaxation. The slow transverse relaxation and narrow line width of the 15N-detected TROSY resonances are expected to compensate for the inherently low 15N sensitivity. However, the sensitivity of 15N-detected TROSY in a previous report was one-order of magnitude lower than in the conventional 1H-detected version. This could be due to the fact that the previous experiments were performed at low salt (0â??50Â*mM), which is advantageous for 1H-detected experiments. Here, we show that the sensitivity gap between 15N and 1H becomes marginal for a non-deuterated, large protein (Ï? cÂ*=Â*35Â*ns) at a physiological salt concentration (200Â*mM). This effect is due to the high salt tolerance of the 15N-detected TROSY. Together with the previously reported benefits of the 15N-detected TROSY, our results provide further support for the significance of this experiment for structural studies of macromolecules when using high field magnets near and above 1Â*GHz.
[NMR paper] Nitrogen detected TROSY at high field yields high resolution and sensitivity for protein NMR.
Nitrogen detected TROSY at high field yields high resolution and sensitivity for protein NMR.
Related Articles Nitrogen detected TROSY at high field yields high resolution and sensitivity for protein NMR.
J Biomol NMR. 2015 Oct 23;
Authors: Takeuchi K, Arthanari H, Shimada I, Wagner G
Abstract
Detection of (15)N in multidimensional NMR experiments of proteins has sparsely been utilized because of the low gyromagnetic ratio (?) of nitrogen and the presumed low sensitivity of such experiments. Here we show that selecting the TROSY...
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10-27-2015 12:33 PM
Nitrogen detected TROSY at high field yields high resolution and sensitivity for protein NMR
Nitrogen detected TROSY at high field yields high resolution and sensitivity for protein NMR
Abstract
Detection of 15N in multidimensional NMR experiments of proteins has sparsely been utilized because of the low gyromagnetic ratio (γ) of nitrogen and the presumed low sensitivity of such experiments. Here we show that selecting the TROSY components of proton-attached 15N nuclei (TROSY 15NH) yields high quality spectra in high field magnets (>600Â*MHz) by taking advantage of the slow 15N transverse relaxation and compensating for the inherently low...
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10-24-2015 05:49 AM
Sensitivity and resolution of proton detected spectra of a deuterated protein at 40 and 60Â*kHz magic-angle-spinning
Sensitivity and resolution of proton detected spectra of a deuterated protein at 40 and 60Â*kHz magic-angle-spinning
Abstract
The use of small rotors capable of very fast magic-angle spinning (MAS) in conjunction with proton dilution by perdeuteration and partial reprotonation at exchangeable sites has enabled the acquisition of resolved, proton detected, solid-state NMR spectra on samples of biological macromolecules. The ability to detect the high-gamma protons, instead of carbons or nitrogens, increases sensitivity. In order to achieve...
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[NMR paper] Perspectives for sensitivity enhancement in proton-detected solid-state NMR of highly deuterated proteins by preserving water magnetization.
Perspectives for sensitivity enhancement in proton-detected solid-state NMR of highly deuterated proteins by preserving water magnetization.
Related Articles Perspectives for sensitivity enhancement in proton-detected solid-state NMR of highly deuterated proteins by preserving water magnetization.
J Biomol NMR. 2015 Jan 30;
Authors: Chevelkov V, Xiang S, Giller K, Becker S, Lange A, Reif B
Abstract
In this work, we show how the water flip-back approach that is widely employed in solution-state NMR can be adapted to...
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01-31-2015 04:16 PM
Perspectives for sensitivity enhancement in proton-detected solid-state NMR of highly deuterated proteins by preserving water magnetization
Perspectives for sensitivity enhancement in proton-detected solid-state NMR of highly deuterated proteins by preserving water magnetization
Abstract
In this work, we show how the water flip-back approach that is widely employed in solution-state NMR can be adapted to proton-detected MAS solid-state NMR of highly deuterated proteins. The scheme allows to enhance the sensitivity of the experiment by decreasing the recovery time of the proton longitudinal magnetization. The method relies on polarization transfer from non-saturated water to the protein...
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01-30-2015 12:15 PM
[NMR paper] Proton-detected MAS NMR experiments based on dipolar transfers for backbone assignment of highly deuterated proteins.
Proton-detected MAS NMR experiments based on dipolar transfers for backbone assignment of highly deuterated proteins.
Related Articles Proton-detected MAS NMR experiments based on dipolar transfers for backbone assignment of highly deuterated proteins.
J Magn Reson. 2014 Mar 4;242C:180-188
Authors: Chevelkov V, Habenstein B, Loquet A, Giller K, Becker S, Lange A
Abstract
Proton-detected solid-state NMR was applied to a highly deuterated insoluble, non-crystalline biological assembly, the Salmonella typhimurium type iii secretion system...
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03-29-2014 01:00 PM
Proton-detected MAS NMR experiments based on dipolar transfers for backbone assignment of highly deuterated proteins
Proton-detected MAS NMR experiments based on dipolar transfers for backbone assignment of highly deuterated proteins
Publication date: Available online 4 March 2014
Source:Journal of Magnetic Resonance</br>
Author(s): Veniamin Chevelkov , Birgit Habenstein , Antoine Loquet , Karin Giller , Stefan Becker , Adam Lange</br>
Proton-detected solid-state NMR was applied to a highly deuterated insoluble, non-crystalline biological assembly, the Salmonella typhimurium type iii secretion system (T3SS) needle. Spectra of very high resolution and sensitivity were obtained...
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03-04-2014 06:37 PM
TROSY NMR Spectroscopy of Large Soluble Proteins.
TROSY NMR Spectroscopy of Large Soluble Proteins.
TROSY NMR Spectroscopy of Large Soluble Proteins.
Top Curr Chem. 2011 Sep 17;
Authors: Xu Y, Matthews S
Abstract
Solution nuclear magnetic resonance spectroscopy is usually only used to study proteins with molecular weight not exceeding about 50 kDa. This size limit has been lifted significantly in recent years, thanks to the development of labelling methods and the application of transverse-relaxation optimized spectroscopy (TROSY). In particular, methyl-specific labelling and...
Nitrogen-detected TROSY yields comparable sensitivity to proton-detected TROSY for non-deuterated, large proteins under physiological salt conditions
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