Peak overlap in crowded regions of two-dimensional spectra prevents characterization of dynamics for many sites of interest in globular and intrinsically disordered proteins. We present new three-dimensional pulse sequences for measurement of Carr-Purcell-Meiboom-Gill relaxation dispersions at backbone nitrogen and carbonyl positions. To alleviate increase in the measurement time associated with the additional spectral dimension, we use non-uniform sampling in combination with two distinct methods of spectrum reconstruction: compressed sensing and co-processing with multi-dimensional decomposition. The new methodology was validated using disordered protein CD79A from B-cell receptor and an SH3 domain from Abp1p in exchange between its free form and bound to a peptide from the protein Ark1p. We show that, while providing much better resolution, the 3D NUS experiments give the similar accuracy and precision of the dynamic parameters to ones obtained using traditional 2D experiments. Furthermore, we show that jackknife resampling of the spectra yields robust estimates of peak intensities errors, eliminating the need for recording duplicate data points.
[NMR paper] Measurement of Ligand-Target Residence Times by (1)H Relaxation Dispersion NMR Spectroscopy.
Measurement of Ligand-Target Residence Times by (1)H Relaxation Dispersion NMR Spectroscopy.
Related Articles Measurement of Ligand-Target Residence Times by (1)H Relaxation Dispersion NMR Spectroscopy.
J Med Chem. 2016 Dec 08;59(23):10788-10793
Authors: Moschen T, Grutsch S, Juen MA, Wunderlich CH, Kreutz C, Tollinger M
Abstract
A ligand-observed (1)H NMR relaxation experiment is introduced for measuring the binding kinetics of low-molecular-weight compounds to their biomolecular targets. We show that this approach, which does...
nmrlearner
Journal club
0
12-11-2016 06:23 AM
Quantitative measurement of exchange dynamics in proteins via 13 C relaxation dispersion of 13 CHD 2 -labeled samples
Quantitative measurement of exchange dynamics in proteins via 13 C relaxation dispersion of 13 CHD 2 -labeled samples
Abstract
Methyl groups have emerged as powerful probes of protein dynamics with timescales from picoseconds to seconds. Typically, studies involving high molecular weight complexes exploit 13CH3- or 13CHD2-labeling in otherwise highly deuterated proteins. The 13CHD2 label offers the unique advantage of providing 13C, 1H and 2H spin probes, however a disadvantage has been the lack of an experiment to record 13C...
nmrlearner
Journal club
0
06-02-2016 02:11 AM
[NMR paper] High resolution NMR study of T? magnetic relaxation dispersion. IV. Proton relaxation in amino acids and Met-enkephalin pentapeptide.
High resolution NMR study of T? magnetic relaxation dispersion. IV. Proton relaxation in amino acids and Met-enkephalin pentapeptide.
http://www.bionmr.com//www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--link.aip.org-jhtml-linkto.gif Related Articles High resolution NMR study of T? magnetic relaxation dispersion. IV. Proton relaxation in amino acids and Met-enkephalin pentapeptide.
J Chem Phys. 2014 Oct 21;141(15):155101
Authors: Pravdivtsev AN, Yurkovskaya AV, Vieth HM, Ivanov KL
Abstract
Nuclear Magnetic Relaxation Dispersion...
nmrlearner
Journal club
0
10-06-2015 10:39 PM
[NMR paper] The Energetics of a Three-State Protein Folding System Probed by High-Pressure Relaxation Dispersion NMR Spectroscopy.
The Energetics of a Three-State Protein Folding System Probed by High-Pressure Relaxation Dispersion NMR Spectroscopy.
Related Articles The Energetics of a Three-State Protein Folding System Probed by High-Pressure Relaxation Dispersion NMR Spectroscopy.
Angew Chem Int Ed Engl. 2015 Sep 14;54(38):11157-11161
Authors: Tugarinov V, Libich DS, Meyer V, Roche J, Clore GM
Abstract
The energetic and volumetric properties of a three-state protein folding system, comprising a metastable triple mutant of the Fyn SH3 domain, have been...
nmrlearner
Journal club
0
09-10-2015 02:01 PM
[Question from NMRWiki Q&A forum] T2 relaxation (CPMG) measurement of 13C high resolution spectra
T2 relaxation (CPMG) measurement of 13C high resolution spectra
Hi all, my question is how to perform a T2 measurement of 13C liquid NMR spectra in BRUKER AVIII ? What sequence to use?
I’ve been try the sequence cpmg, just like in this linkhttp://nmr.chinanmr.cn/guide/tutorials/1Dclas/t2h1.html
My problem is that I want to decouple the spectra… Does anyone know what sequence should I use?
Any help will be appreciated. Thank you
nmrlearner
News from other NMR forums
0
01-30-2014 05:27 AM
Measurement of protein unfolding/refolding kinetics and structural characterization of hidden intermediates by NMR relaxation dispersion [Biophysics and Computational Biology]
Measurement of protein unfolding/refolding kinetics and structural characterization of hidden intermediates by NMR relaxation dispersion
Meinhold, D. W., Wright, P. E....
Date: 2011-05-31
Detailed understanding of protein function and malfunction hinges on the ability to characterize transiently populated states and the transitions between them. Here, we use 15N, , and 13CO NMR R2 relaxation dispersion to investigate spontaneous unfolding and refolding events of native apomyoglobin. Above pH 5.0, dispersion is dominated by processes involving fluctuations of the F-helix region, which...
nmrlearner
Journal club
0
05-31-2011 11:41 PM
Measurement of protein unfolding/refolding kinetics and structural characterization of hidden intermediates by NMR relaxation dispersion.
Measurement of protein unfolding/refolding kinetics and structural characterization of hidden intermediates by NMR relaxation dispersion.
Measurement of protein unfolding/refolding kinetics and structural characterization of hidden intermediates by NMR relaxation dispersion.
Proc Natl Acad Sci U S A. 2011 May 11;
Authors: Meinhold DW, Wright PE
Detailed understanding of protein function and malfunction hinges on the ability to characterize transiently populated states and the transitions between them. Here, we use (15)N, , and (13)CO NMR R(2)...
nmrlearner
Journal club
0
05-13-2011 02:40 PM
Measurement of carbonyl chemical shifts of excited protein states by relaxation dispersion NMR spectroscopy: comparison between uniformly and selectively 13C labeled samples
Measurement of carbonyl chemical shifts of excited protein states by relaxation dispersion NMR spectroscopy: comparison between uniformly and selectively 13C labeled samples
Patrik Lundström, D. Flemming Hansen and Lewis E. Kay
Journal of Biomolecular NMR; 2008; 42(1); pp 35 - 47
Abstract: Carr–Purcell–Meiboom–Gill (CPMG) relaxation dispersion nuclear magnetic resonance (NMR) spectroscopy has emerged as a powerful method for quantifying chemical shifts of excited protein states. For many applications of the technique that involve the measurement of relaxation rates of carbon...
Measurement of protein backbone 13 CO and 15 N relaxation dispersion at high resolution
Linear Mode
