Publication date: Available online 3 May 2016 Source:Journal of Magnetic Resonance
Author(s): Julien Roche, Jinfa Ying, Yang Shen, Dennis A. Torchia, Ad Bax
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 during a delay, integrated into the regular two-dimensional TROSY-HSQC pulse scheme, and compares the obtained intensity with a reference spectrum where 3 J HNH? dephasing is suppressed. The effect of passive 1H? spin flips downscales the apparent 3 J HNH? coupling by a uniform factor that depends approximately linearly on both the duration of the 3 J HNH? dephasing delay and the 1H-1H cross relaxation rate. Using such a correction factor, which accounts for the effects of both inhomogeneity of the radiofrequency field and 1H? spin flips, agreement between prior and newly measured values for the small model protein GB3 is better than 0.3 Hz. Measurement for the HIV protease homodimer (22 kDa) yields 3 J HNH? values that agree to better than 0.7 Hz with predictions made on the basis of a previously parameterized Karplus equation. Although for Gly residues the two individual 3 J HNH? couplings cannot be extracted from a single set of ARTSY-J spectra, the measurement provides valuable ? angle information. Graphical abstract
An encodable lanthanide binding tag with reduced size and flexibility for measuring residual dipolar couplings and pseudocontact shifts in large proteins
An encodable lanthanide binding tag with reduced size and flexibility for measuring residual dipolar couplings and pseudocontact shifts in large proteins
Abstract
Metal ions serve important roles in structural biology applications from long-range perturbations seen in magnetic resonance experiments to electron-dense signatures in X-ray crystallography data; however, the metal ion must be secured in a molecular framework to achieve the maximum benefit. Polypeptide-based lanthanide-binding tags (LBTs) represent one option that can be directly encoded...
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01-04-2016 07:49 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
A PositivelyCharged Liquid Crystalline Medium forMeasuring Residual Dipolar Couplings in Membrane Proteins by NMR
A PositivelyCharged Liquid Crystalline Medium forMeasuring Residual Dipolar Couplings in Membrane Proteins by NMR
Pallavi Thiagarajan-Rosenkranz, Adrian W. Draney, Sean T. Smrt and Justin L. Lorieau
http://pubs.acs.org/appl/literatum/publisher/achs/journals/content/jacsat/0/jacsat.ahead-of-print/jacs.5b07515/20150910/images/medium/ja-2015-07515m_0004.gif
Journal of the American Chemical Society
DOI: 10.1021/jacs.5b07515
http://feeds.feedburner.com/~ff/acs/jacsat?d=yIl2AUoC8zA
http://feeds.feedburner.com/~r/acs/jacsat/~4/Q6RR1Z4u0gw
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09-11-2015 06:48 AM
[NMR paper] A Positively Charged Liquid Crystalline Medium for Measuring Residual Dipolar Couplings in Membrane Proteins by NMR.
A Positively Charged Liquid Crystalline Medium for Measuring Residual Dipolar Couplings in Membrane Proteins by NMR.
A Positively Charged Liquid Crystalline Medium for Measuring Residual Dipolar Couplings in Membrane Proteins by NMR.
J Am Chem Soc. 2015 Sep 8;
Authors: Thiagarajan-Rosenkranz Paltr Uic Edu P, Draney AW, Smrt ST, Lorieau JL
Abstract
Residual Dipolar Couplings (RDCs) are integral to the refinement of membrane protein structures by NMR since they accurately define the orientation of helices and other structural...
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09-09-2015 11:49 AM
Highly Precise Measurementof Kinetic Isotope EffectsUsing 1H-Detected 2D [13C,1H]-HSQC NMR Spectroscopy
Highly Precise Measurementof Kinetic Isotope EffectsUsing 1H-Detected 2D -HSQC NMR Spectroscopy
Kathryn A. Manning, Bharathwaj Sathyamoorthy, Alexander Eletsky, Thomas Szyperski and Andrew S. Murkin
http://pubs.acs.org/appl/literatum/publisher/achs/journals/content/jacsat/0/jacsat.ahead-of-print/ja310353c/aop/images/medium/ja-2012-10353c_0005.gif
Journal of the American Chemical Society
DOI: 10.1021/ja310353c
http://feeds.feedburner.com/~ff/acs/jacsat?d=yIl2AUoC8zA
http://feeds.feedburner.com/~r/acs/jacsat/~4/kfh2aVSYThA
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12-12-2012 08:19 AM
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
Measurement of 1Hâ??15N and 1Hâ??13C residual dipolar couplings in nucleic acids from TROSY intensities
Measurement of 1Hâ??15N and 1Hâ??13C residual dipolar couplings in nucleic acids from TROSY intensities
Abstract Analogous to the recently introduced ARTSY method for measurement of one-bond 1Hâ??15N residual dipolar couplings (RDCs) in large perdeuterated proteins, we introduce methods for measurement of base 13Câ??1H and 15Nâ??1H RDCs in protonated nucleic acids. Measurements are based on quantitative analysis of intensities in 1Hâ??15N and 13Câ??1H TROSY-HSQC spectra, and are illustrated for a 71-nucleotide adenine riboswitch. Results compare favorably with those of conventional...
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09-30-2011 08:01 PM
MQ-HNCO-TROSY for the measurement of scalar and residual dipolar couplings in larger
Abstract We describe a novel pulse sequence, MQ-HNCO-TROSY, for the measurement of scalar and residual dipolar couplings between amide proton and nitrogen in larger proteins. The experiment utilizes the whole 2TN polarization transfer delay for labeling of 15N chemical shift in a constant time manner, which efficiently doubles the attainable resolution in 15N dimension with respect to the conventional HNCO-TROSY experiment. In addition, the accordion principle is employed for measuring (J + D)NHs, and the multiplet components are selected with the generalized version of the TROSY scheme...
ARTSY-J: Convenient and precise measurement of 3JHNH? couplings in medium-size proteins from TROSY-HSQC spectra
Linear Mode
