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.
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McIntosh, L.P., Brun, E. & Kay, L.E. Stereospecific assignment of the NH2 resonances from the primary amides of asparagine and glutamine side chains in isotopically labeled proteins. J Biomol NMR 9, 306–312 (1997). https://doi.org/10.1023/A:1018635110491
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DOI: https://doi.org/10.1023/A:1018635110491