Determination of all NOes in 1H–13C–Me-ILV-U−2H–15N Proteins with Two Time-Shared Experiments

Authors

Dominique P. Frueh, Harvard University
David A. Vosburg, Harvey Mudd CollegeFollow
Christopher T. Walsh, Harvard University
Gerhard Wagner, Harvard University

Document Type

Article

Department

Chemistry (HMC)

Publication Date

2006

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.

Rights Information

© 2006 Springer

DOI

10.1007/s10858-005-5338-4

Recommended Citation

“Determination of all nOes in 1H-13C-Me-ILV-U-2H-15N proteins with two time-shared experiments,” Frueh, D.P.; Vosburg, D.A.; Walsh, C.T.; Wagner, G. J. Biomol. NMR 2006, 34, 31-40. DOI: 10.1007/s10858-005-5338-4