A TROSY-based NMR experiment is described for simultaneous measurement of the 15N longitudinal relaxation rate constant R1 and the {1H}â??15N nuclear Overhauser enhancement. The experiment is based on the observation that the TROSY mixing pulse sequence element symmetrically exchanges 1H and 15N magnetizations. The accuracy of the proposed technique is validated by comparison to independent measurements of both relaxation parameters for the protein ubiquitin. The simultaneous experiment is approximately 20â??33% shorter than conventional sequential measurements.
[NMR paper] ARTSY-J: Convenient and precise measurement of 3JHNH? couplings in medium-size proteins from TROSY-HSQC spectra
ARTSY-J: Convenient and precise measurement of 3JHNH? couplings in medium-size proteins from TROSY-HSQC spectra
Publication date: Available online 3 May 2016
Source:Journal of Magnetic Resonance</br>
Author(s): Julien Roche, Jinfa Ying, Yang Shen, Dennis A. Torchia, Ad Bax</br>
A new and convenient method, named ARTSY-J, is introduced that permits extraction of the 3 J HNH? couplings in proteins from the relative intensities in a pair of 15N-1H TROSY-HSQC spectra. The pulse scheme includes 3 J HNH? dephasing of the narrower TROSY 1HN-{15N} doublet component...
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05-05-2016 03:15 AM
Nitrogen-detected TROSY yields comparable sensitivity to proton-detected TROSY for non-deuterated, large proteins under physiological salt conditions
Nitrogen-detected TROSY yields comparable sensitivity to proton-detected TROSY for non-deuterated, large proteins under physiological salt conditions
Abstract
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...
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01-23-2016 03:35 PM
Accurate measurement of 3 J HNHα couplings in small or disordered proteins from WATERGATE-optimized TROSY spectra
Accurate measurement of 3 J HNHα couplings in small or disordered proteins from WATERGATE-optimized TROSY spectra
Abstract
Provided that care is taken in adjusting the WATERGATE element of a 1Hâ??15N TROSY-HSQC experiment, such that neither the water magnetization nor the 1Hα protons are inverted by its final 180° pulse, 3JHNHα couplings can be measured directly from splittings in the 1H dimension of the spectrum. With band-selective 1H decoupling, very high 15N resolution can be achieved. A complete set of 3JHNHα values, ranging from 3.4 to...
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12-10-2015 05:49 PM
[NMR paper] MAS solid state NMR of proteins: simultaneous (15)N- (13)CA and (15)N- (13)CO dipolar recoupling via low-power symmetry-based RF pulse schemes.
MAS solid state NMR of proteins: simultaneous (15)N- (13)CA and (15)N- (13)CO dipolar recoupling via low-power symmetry-based RF pulse schemes.
MAS solid state NMR of proteins: simultaneous (15)N- (13)CA and (15)N- (13)CO dipolar recoupling via low-power symmetry-based RF pulse schemes.
J Biomol NMR. 2015 Feb 25;
Authors: Herbst C, Bellstedt P, Görlach M, Ramachandran R
Abstract
The generation of efficient RN n (?)s,(?)k symmetry-based low-power RF pulse schemes for simultaneous (15)N-(13)CA and (15)N-(13)CO dipolar recoupling...
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02-26-2015 11:11 PM
MAS solid state NMR of proteins: simultaneous 15 Nâ?? 13 CA and 15 Nâ?? 13 CO dipolar recoupling via low-power symmetry-based RF pulse schemes
MAS solid state NMR of proteins: simultaneous 15 Nâ?? 13 CA and 15 Nâ?? 13 CO dipolar recoupling via low-power symmetry-based RF pulse schemes
Abstract
The generation of efficient RN n νs,νk symmetry-based low-power RF pulse schemes for simultaneous 15Nâ??13CA and 15Nâ??13CO dipolar recoupling is demonstrated. The method involves mixing schemes employing phase and amplitude-modulated dual band-selective 180° pulses as basic â??Râ?? element and tailoring of the RF field-modulation...
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02-25-2015 05:56 PM
[NMR paper] Synthesis of uniformly deuterated n-dodecyl-?-D-maltoside (d39 -DDM) for solubilization of membrane proteins in TROSY NMR experiments.
Synthesis of uniformly deuterated n-dodecyl-?-D-maltoside (d39 -DDM) for solubilization of membrane proteins in TROSY NMR experiments.
Related Articles Synthesis of uniformly deuterated n-dodecyl-?-D-maltoside (d39 -DDM) for solubilization of membrane proteins in TROSY NMR experiments.
J Labelled Comp Radiopharm. 2014 Dec 9;
Authors: Hiruma-Shimizu K, Kalverda AP, Henderson PJ, Homans SW, Patching SG
Abstract
This work reports the first synthesis of uniformly deuterated n-dodecyl-?-D-maltoside (d39 -DDM). DDM is a mild non-ionic...
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12-11-2014 11:22 PM
Measurement of 15N relaxation rates in perdeuterated proteins by TROSY-based methods
Measurement of 15N relaxation rates in perdeuterated proteins by TROSY-based methods
<div class="Abstract" lang="en">Abstract <div class="normal">While extracting dynamics parameters from backbone 15N relaxation measurements in proteins has become routine over the past two decades, it is increasingly recognized that accurate quantitative analysis can remain limited by the potential presence of systematic errors associated with the measurement of 15N R1 and R2 or R1Ï? relaxation rates as well as heteronuclear 15N-{1H} NOE values. We show that systematic errors in such measurements can...
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06-16-2012 06:01 AM
Simultaneous measurement of 1Hâ??15N and Methyl 1Hmâ??13Cm residual dipolar couplings in large proteins
Simultaneous measurement of 1Hâ??15N and Methyl 1Hmâ??13Cm residual dipolar couplings in large proteins
Abstract A two-dimensional TROSY-based SIM-13Cmâ??1Hm/1Hâ??15N NMR experiment for simultaneous measurements of methyl 1 D CH and backbone amide 1 D NH residual dipolar couplings (RDC) in {U-; Ileδ1-; Leu,Val-}-labeled samples of large proteins is described. Significant variation in the alignment tensor of the 82-kDa enzyme Malate synthase G is observed as a function of only slight changes in experimental conditions. The SIM-13Cmâ??1Hm/1Hâ??15N data sets provide convenient means...
TROSY pulse sequence for simultaneous measurement of the 15 N R 1 and { 1 H}â?? 15 N NOE in deuterated proteins
Linear Mode
