Simultaneous measurement of protein one-bond and two-bond nitrogen-carbon coupling constants using an internally referenced quantitative J-correlated [15N,1H]-TROSY-HNC experiment

Wienk, Hans L.J. and Martinez, Mitcheell M. and Yalloway, Gary N. and Schmidt, Jürgen M. and Perez, Carlos S and Rüterjans, Heinz and Löhr, Frank (2003) Simultaneous measurement of protein one-bond and two-bond nitrogen-carbon coupling constants using an internally referenced quantitative J-correlated [15N,1H]-TROSY-HNC experiment. Journal of Biomolecular NMR, 25 (2). pp. 133-145. ISSN 0925-2738 (Print) 1573-5001 (Online). (The full text of this publication is not currently available from this repository. You may be able to access a copy if URLs are provided)

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Official URL
http://dx.doi.org/10.1023/A:1022233103990

Abstract

A quantitative J-correlation pulse sequence is described that allows simultaneous determination of one-bond and two-bond nitrogen-carbon coupling constants for protonated or deuterated proteins. Coupling constants are calculated from volume ratios between cross peaks and reference axial peaks observed in a single 3D spectrum. Accurate backbone 1JNC', 1JNCα, and 2JNCα coupling constants are obtained for the two [15N;13C]-labeled, medium-sized proteins flavodoxin and xylanase and for the [2H;15N;13C]-labeled, large protein DFPase. A dependence of onebond and two-bond JNCα values on protein backbone ψ torsion angles is readily apparent, in agreement with previously found correlations. In addition, the experiment is performed on isotropic as well as aligned protein to measure associated 15N-13C residual dipolar couplings.

Item Type: Article
Uncontrolled keywords: DFPase, flavodoxin, perdeuteration, residual dipolar coupling, scalar coupling, TROSY, xylanase
Subjects: Q Science
Q Science > QD Chemistry
Q Science > QP Physiology (Living systems)
Q Science > QC Physics
Q Science > QP Physiology (Living systems) > QP506 Molecular biology
Divisions: Faculties > Science Technology and Medical Studies > School of Biosciences > Protein Science Group
Depositing User: Jurgen M Schmidt
Date Deposited: 30 Sep 2009 15:56
Last Modified: 06 Jun 2014 11:33
Resource URI: https://kar.kent.ac.uk/id/eprint/22839 (The current URI for this page, for reference purposes)
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