Structure determination of proteins by solution NMR has become an established method, but challenges increase steeply with the size of proteins. Notably, spectral crowding and signal overlap impair the analysis of cross-peaks in NOESY spectra that provide distance restraints for structural models. An optimal spectral resolution can alleviate overlap but requires prohibitively long experimental time with existing methods. Here we present a time-shared 3D experiment optimized for large proteins that provides 15N and 13C dispersed NOESY spectra in a single measurement. NOESY correlations appear in the detected dimension and hence benefit from the highest resolution achievable of all dimensions without increase in experimental time. By design, this experiment is inherently optimal for non-uniform sampling acquisition when compared to current alternatives. Thus, 15N and 13C dispersed NOESY spectra with ultra-high resolution in all dimensions were acquired in parallel within about 4Â*days instead of 80Â*days for a 52Â*kDa monomeric protein at a concentration of 350Â*μM.
[NMR paper] A 3D time-shared NOESY experiment designed to provide optimal resolution for accurate assignment of NMR distance restraints in large proteins.
A 3D time-shared NOESY experiment designed to provide optimal resolution for accurate assignment of NMR distance restraints in large proteins.
Related Articles A 3D time-shared NOESY experiment designed to provide optimal resolution for accurate assignment of NMR distance restraints in large proteins.
J Biomol NMR. 2014 Nov 9;
Authors: Mishra SH, Harden BJ, Frueh DP
Abstract
Structure determination of proteins by solution NMR has become an established method, but challenges increase steeply with the size of proteins. Notably,...
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11-10-2014 10:59 PM
Time-shared experiments for efficient assignment of triple-selectively labeled proteins
Time-shared experiments for efficient assignment of triple-selectively labeled proteins
Publication date: Available online 30 September 2014
Source:Journal of Magnetic Resonance</br>
Author(s): Frank Löhr , Aisha Laguerre , Christoph Bock , Sina Reckel , Peter J. Connolly , Norzehan Abdul-Manan , Franz Tumulka , Rupert Abele , Jonathan M. Moore , Volker Dötsch</br>
Combinatorial triple-selective labeling facilitates the NMR assignment process for proteins that are subject to signal overlap and insufficient signal-to-noise in standard triple-resonance...
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10-01-2014 02:48 AM
Efficient Acquisition of High-Resolution 4-D Diagonal-Suppressed Methyl-Methyl NOESY for Large Proteins
Efficient Acquisition of High-Resolution 4-D Diagonal-Suppressed Methyl-Methyl NOESY for Large Proteins
Publication year: 2012
Source:Journal of Magnetic Resonance</br>
Jie Wen, Jihui Wu, Pei Zhou</br>
The methyl-methyl NOESYexperimentplays an important role in determiningthe global folds of large proteins. Despite the high sensitivity of this experiment, the analysisof methyl-methyl NOEs is frequently hindered by the limited chemical shift dispersion of methyl groups, particularly methyl protons. Thismakes it difficult to unambiguously assign all of the methyl-methyl...
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03-10-2012 10:54 AM
Time-shared HSQC-NOESY for accurate distance constraints measured at high-field in 15N-13C-ILV methyl labeled proteins
Time-shared HSQC-NOESY for accurate distance constraints measured at high-field in 15N-13C-ILV methyl labeled proteins
Abstract We present a time-shared 3D HSQC-NOESY experiment that enables one to simultaneously record 13C- and 15N-dispersed spectra in Ile, Leu and Val (ILV) methyl-labeled samples. This experiment is designed to delineate the two spectra which would otherwise overlap with one another when acquired together. These spectra display nOe correlations in the detected proton dimension, i.e. with maximum resolution. This is in contrast to NOESY-HSQC types of experiments that...
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[NMR paper] Automated NOESY interpretation with ambiguous distance restraints: the refined NMR so
Automated NOESY interpretation with ambiguous distance restraints: the refined NMR solution structure of the pleckstrin homology domain from beta-spectrin.
http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--linkinghub.elsevier.com-ihub-images-PubMedLink.gif Related Articles Automated NOESY interpretation with ambiguous distance restraints: the refined NMR solution structure of the pleckstrin homology domain from beta-spectrin.
J Mol Biol. 1997 Jun 13;269(3):408-22
Authors: Nilges M, Macias MJ, O'Donoghue SI, Oschkinat H
We have used a...
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[NMR paper] Automated NOESY interpretation with ambiguous distance restraints: the refined NMR so
Automated NOESY interpretation with ambiguous distance restraints: the refined NMR solution structure of the pleckstrin homology domain from beta-spectrin.
http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--linkinghub.elsevier.com-ihub-images-PubMedLink.gif Related Articles Automated NOESY interpretation with ambiguous distance restraints: the refined NMR solution structure of the pleckstrin homology domain from beta-spectrin.
J Mol Biol. 1997 Jun 13;269(3):408-22
Authors: Nilges M, Macias MJ, O'Donoghue SI, Oschkinat H
We have used a...
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08-22-2010 03:03 PM
[NMR paper] Proton NMR studies of a large protein. pH, substrate titrations, and NOESY experiment
Proton NMR studies of a large protein. pH, substrate titrations, and NOESY experiments with perdeuterated yeast phosphoglycerate kinase containing histidine residues.
Related Articles Proton NMR studies of a large protein. pH, substrate titrations, and NOESY experiments with perdeuterated yeast phosphoglycerate kinase containing histidine residues.
J Magn Reson B. 1994 Oct;105(2):157-66
Authors: Pappu KM, Serpersu EH
Fully deuterated yeast phosphoglycerate kinase (PGK) was prepared biosynthetically with only histidine side chains of normal...
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Simultaneous detection of amide and methyl correlations using a time shared NMR experiment: application to binding epitope mapping
Simultaneous detection of amide and methyl correlations using a time shared NMR experiment: application to binding epitope mapping
Peter Würtz, Olli Aitio, Maarit Hellman and Perttu Permi
Journal of Biomolecular NMR; 2007; 39(2) pp 97 - 105
Abstract:
Simultaneous recording of different NMR parameters is an efficient way to reduce the overall experimental time and speed up structural studies of biological macromolecules. This can especially be beneficial in the case of fast NMR-based drug screening applications or for collecting NOE restraints, where prohibitively long data collection...
A 3D time-shared NOESY experiment designed to provide optimal resolution for accurate assignment of NMR distance restraints in large proteins
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