Journal of Biomolecular NMR

, Volume 9, Issue 3, pp 306–312

Stereospecific assignment of the NH2 resonances from the primary amides of asparagine and glutamine side chains in isotopically labeled proteins

  • Lawrence P. McIntosh
  • Emmanuel Brun
  • Lewis E. Kay
Article

DOI: 10.1023/A:1018635110491

Cite this article as:
McIntosh, L.P., Brun, E. & Kay, L.E. J Biomol NMR (1997) 9: 306. doi:10.1023/A:1018635110491

Abstract

An HMQC-based pulse scheme is presented for the stereospecific assignment of asparagineand glutamine side-chain amide protons. The approach makes use of the recently developedquantitative-J correlation spectroscopy [Bax, A. et al. (1994) Methods Enzymol., 239,79–105] to distinguish the E and Z primary amide protons and, as such, eliminates theneed for assignments derived from more time-consuming and potentially ambiguous NOEmethods. An application of this method to a uniformly 15N,13C-labeled cellulose-bindingdomain is presented. When used in combination with a NOESY-HSQC experiment, thepredominant χ2 dihedral angles of two asparagine side chains in this protein can also bedefined.

Primary amide Stereospecific assignment HMQC spectroscopy Dihedral angle 

Copyright information

© Kluwer Academic Publishers 1997

Authors and Affiliations

  • Lawrence P. McIntosh
    • 1
  • Emmanuel Brun
    • 1
  • Lewis E. Kay
    • 2
  1. 1.Protein Engineering Network Centers of Excellence, Department of Biochemistry and Molecular Biology and Department of ChemistryUniversity of British ColumbiaVancouverCanada
  2. 2.Protein Engineering Network Centers of Excellence and Departments of Medical Genetics, Biochemistry and ChemistryUniversity of TorontoTorontoCanada

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