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Improved method for unambiguous amino acid side‐chain 1 H and 13 C resonance assignment
Author(s) -
Löhr Frank,
Betz Marco,
Rüterjans Heinz
Publication year - 2004
Publication title -
magnetic resonance in chemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.483
H-Index - 72
eISSN - 1097-458X
pISSN - 0749-1581
DOI - 10.1002/mrc.1346
Subject(s) - chemistry , resonance (particle physics) , side chain , stereochemistry , atomic physics , organic chemistry , physics , polymer
Side‐chain proton and carbon‐13 resonance assignments of [ 13 C; 15 N]‐enriched proteins usually rely on combinations of several multi‐dimensional experiments. Here, we describe a four‐dimensional pulse sequence, H(C)C–COSY–TOCSY–(CACO)NH, which provides the information required to assign completely aliphatic side‐chain resonance frequencies. As in widely used HCC(CO)NH–TOCSY experiments, problems due to spectral crowding are alleviated by exploiting the dispersion of backbone amide 1 H and 15 N signals. The modification introduced here allows signals from different side‐chains to be distinguished even in the case of overlap in the 1 H N – 15 N plane of the spectra. For illustration, the new method is applied to two proteins with molecular masses of 11 and 23 kDa. Copyright © 2004 John Wiley & Sons, Ltd.