A simple method for measuring signs of 1HN chemical shift differences between ground
Abstract NMR relaxation dispersion spectroscopy is a powerful method for studying protein conformational dynamics whereby visible, ground and invisible, excited conformers interconvert on the millisecond time-scale. In addition to providing kinetics and thermodynamics parameters of the exchange process, the CPMG dispersion experiment also allows extraction of the absolute values of the chemical shift differences between interconverting states,
ê
ê \Updelta ~
w
ê
ê , opening the way for structure determination of excited state conformers. Central to the goal of structural analysis is the availability of the chemical shifts of the excited state that can only be obtained once the signs of
\Updelta ~
w
are known. Herein we describe a very simple method for determining the signs of 1HN
\Updelta ~
w
values based on a comparison of peak positions in the directly detected dimensions of a pair of 1HNâ??15N correlation maps recorded at different static magnetic fields. The utility of the approach is demonstrated for three proteins that undergo millisecond time-scale conformational rearrangements. Although the method provides fewer signs than previously published techniques it does have a number of strengths: (1) Data sets needed for analysis are typically available from other experiments, such as those required for measuring signs of 15N
\Updelta ~
w
values, thus requiring no additional experimental time, (2) acquisition times in the critical detection dimension can be as long as necessary and (3) the signs obtained can be used to cross-validate those from other approaches.
Content Type Journal Article
DOI 10.1007/s10858-010-9418-8
Authors
Guillaume Bouvignies, The University of Toronto Departments of Molecular Genetics, Biochemistry and Chemistry Toronto Ontario M5S 1A8 Canada
Dmitry M. Korzhnev, The University of Toronto Departments of Molecular Genetics, Biochemistry and Chemistry Toronto Ontario M5S 1A8 Canada
Philipp Neudecker, The University of Toronto Departments of Molecular Genetics, Biochemistry and Chemistry Toronto Ontario M5S 1A8 Canada
D. Flemming Hansen, The University of Toronto Departments of Molecular Genetics, Biochemistry and Chemistry Toronto Ontario M5S 1A8 Canada
Matthew H. J. Cordes, The University of Arizona Department of Chemistry and Biochemistry Tucson AZ 85721-0088 USA
Lewis E. Kay, The University of Toronto Departments of Molecular Genetics, Biochemistry and Chemistry Toronto Ontario M5S 1A8 Canada
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Abstract Carr-Purcell-Meiboom-Gill (CPMG) relaxation dispersion NMR spectroscopy has emerged as a powerful tool for quantifying the kinetics and thermodynamics of millisecond exchange processes between a major, populated ground state and one or more minor, low populated and often invisible â??excitedâ?? conformers. Analysis of CPMG data-sets also provides the magnitudes of the chemical shift difference(s) between exchanging states (|Î?Ï?|), that inform on the structural properties of the excited state(s). The sign of Î?Ï? is, however, not available from CPMG data. Here we present...
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A simple and user-friendly method of labeling protein selectively with amino acids in vivo is introduced. This technique does not require the use of transaminase-deficient or auxotrophic strains. By manipulating the product feedback inhibitory loops of the E. coli amino acid metabolic pathways and, if necessary, by using enzyme inhibitors, proteins were labeled efficiently in vivo...
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Chemical shift of protons?
Can someone give me the answers to the chemical shift of the protons in this compound: CH3CH2OCH2CH(CH3)COOH? Of if you can explain how to get the chemical shift it would be quite helpful as the info online does not really help me.
A simple method for measuring signs of 1HN chemical shift differences between ground
Linear Mode
