Determination of all nOes in 1H-13C-Me-ILV-U-2H-15N proteins with two time-shared experiments

Journal of Biomolecular NMR
Dominique P FruehGerhard Wagner

Abstract

We present two time-shared experiments that enable the characterization of all nOes in 1H-13C-ILV methyl-labelled proteins that are otherwise uniformly deuterated and 15N enriched and possibly selectively protonated for distinct residue types. A 3D experiment simultaneously provides the spectra of a 3D NOESY-HN-TROSY and of a 3D NOESY-HC-PEP-HSQC. Thus, nOes from any protons to methyl or amide protons are dispersed with respect to 15N and 13C chemical shifts, respectively. The single 4D experiment presented here yields simultaneously the four 4D experiments HC-HSQC-NOESY-HC-PEP-HSQC, HC-HSQC-NOESY-HN-TROSY, HN-HSQC-NOESY-HN-TROSY and HN-HSQC-NOESY-HC-PEP-HSQC. This allows for the unambiguous determination of all nOes involving amide and methyl protons. The method was applied to a (1H,13C)-ILV-(1H)-FY-(U-2H,15N) sample of a 37 kDa di-domain of the E. coli enterobactin synthetase module EntF.

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Citations

Nov 13, 2008·Journal of Biomolecular NMR·Chenyun Guo, Vitali Tugarinov
Sep 10, 2009·Journal of the American Chemical Society·Sebastian HillerVladislav Y Orekhov
Jul 29, 2009·Natural Product Reports·Alexander Koglin, Christopher T Walsh
Feb 19, 2014·Progress in Nuclear Magnetic Resonance Spectroscopy·Dominique P Frueh
Jun 18, 2014·Journal of the American Chemical Society·Rasmus LinserGerhard Wagner
May 23, 2009·Chemphyschem : a European Journal of Chemical Physics and Physical Chemistry·Isabella C Felli, Bernhard Brutscher
Mar 14, 2007·Magnetic Resonance in Chemistry : MRC·Miriam Pérez-TrujilloTeodor Parella
Jul 16, 2013·Current Opinion in Structural Biology·Dominique P FruehScott R Nichols
Dec 3, 2014·Journal of Magnetic Resonance·Frank LöhrVolker Dötsch
Jun 1, 2007·Journal of the American Chemical Society·Yingqi XuDaiwen Yang
Nov 2, 2007·Journal of the American Chemical Society·David A SnyderRafael Brüschweiler
Jan 18, 2007·Journal of the American Chemical Society·Remy SounierJérôme Boisbouvier

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