Determination of All NOes in 1H-13C-Me-ILV-U-2H-15N Proteins with Two Time-shared Experiments

Overview

Journal J Biomol NMR

Publisher Springer

Specialties Molecular Biology
Nuclear Medicine

Date 2006 Mar 1

PMID 16505962

Citations 15

Authors

Dominique P Frueh

David A Vosburg

Christopher T Walsh

Gerhard Wagner

Affiliations

Soon will be listed here.

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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