Proton detection in solid-state NMR has seen a tremendous increase in popularity in the last years. New experimental techniques allow to exploit protons as an additional source of information on structure, dynamics, and protein interactions with their surroundings. In addition, sensitivity is mostly improved and ambiguity in assignment experiments reduced. We show here that, in the solid state, sequential amide-to-amide correlations turn out to be an excellent, complementary way to exploit amide shifts for unambiguous backbone assignment. For a general assessment, we compare amide-to-amide experiments with the more common 13C-shift-based methods. Exploiting efficient CP magnetization transfers rather than less efficient INEPT periods, our results suggest that the approach is very feasible for solid-state NMR.
[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
[NMR paper] Reduced Dimensionality tailored HN(C)N Experiments for Facile Backbone Resonance Assignment of Proteins through Unambiguous Identification of Sequential HSQC Peaks
Reduced Dimensionality tailored HN(C)N Experiments for Facile Backbone Resonance Assignment of Proteins through Unambiguous Identification of Sequential HSQC Peaks
Publication date: Available online 8 October 2013
Source:Journal of Magnetic Resonance</br>
Author(s): Dinesh Kumar</br>
Two novel reduced dimensionality (RD) tailored HN(C)N experiments are proposed to facilitate the backbone resonance assignment of proteins both in terms of its accuracy and speed. These experiments -referred here as (4,3)D-hNCOcaNH and (4,3)D-hNcoCANH- exploit the linear combination of...
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10-09-2013 05:31 AM
Restraints on backbone conformations in solid state NMR studies of uniformly labeled proteins from quantitative amide 15N–15N and carbonyl 13C–13C dipolar recoupling data
Restraints on backbone conformations in solid state NMR studies of uniformly labeled proteins from quantitative amide 15N–15N and carbonyl 13C–13C dipolar recoupling data
May 2012
Publication year: 2012
Source:Journal of Magnetic Resonance, Volume 218</br>
</br>
Recent structural studies of uniformly 15N, 13C-labeled proteins by solid state nuclear magnetic resonance (NMR) rely principally on two sources of structural restraints: (i) restraints on backbone conformation from isotropic 15N and 13C chemical shifts, based on empirical correlations between chemical shifts and...
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02-03-2013 10:13 AM
Restraints on backbone conformations in solid state NMR studies of uniformly labeled proteins from quantitative amide 15N-15N and carbonyl 13C-13C dipolar recoupling data
Restraints on backbone conformations in solid state NMR studies of uniformly labeled proteins from quantitative amide 15N-15N and carbonyl 13C-13C dipolar recoupling data
Publication year: 2012
Source:Journal of Magnetic Resonance</br>
Kan-Nian Hu, Wei Qiang, Guillermo A. Bermejo, Charles D. Schwieters, Robert Tycko</br>
Recent structural studies of uniformly 15N,13C-labeled proteins by solid state nuclear magnetic resonance (NMR) rely principally on two sources of structural restraints: (i) restraints on backbone conformation from isotropic 15N and 13C chemical...
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03-10-2012 10:54 AM
[NMR paper] A combinatorial selective labeling method for the assignment of backbone amide NMR re
A combinatorial selective labeling method for the assignment of backbone amide NMR resonances.
Related Articles A combinatorial selective labeling method for the assignment of backbone amide NMR resonances.
J Am Chem Soc. 2004 Apr 28;126(16):5020-1
Authors: Parker MJ, Aulton-Jones M, Hounslow AM, Craven CJ
A combinatorial selective labeling (CSL) method is presented for the assignment of backbone amide NMR resonances, which has a particular application in the identification of protein-ligand interaction sites. The method builds on the dual...
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11-24-2010 09:51 PM
13C-direct detected NMR experiments for the sequential J-based resonance assignment o
Abstract We present here a set of 13C-direct detected NMR experiments to facilitate the resonance assignment of RNA oligonucleotides. Three experiments have been developed: (1) the (H)CC-TOCSY-experiment utilizing a virtual decoupling scheme to assign the intraresidual ribose 13C-spins, (2) the (H)CPC-experiment that correlates each phosphorus with the C4â?² nuclei of adjacent nucleotides via J(C,P) couplings and (3) the (H)CPC-CCH-TOCSY-experiment that correlates the phosphorus nuclei with the respective C1â?²,H1â?² ribose signals. The experiments were applied to two RNA hairpin structures....
Sequential backbone assignment based on dipolar amide-to-amide correlation experiments
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