Data requirements for reliable chemical shift assignments in deuterated proteins.
Date of Original Version
Abstract or Description
The information required for chemical shift assignments in large deuterated proteins was investigated using a Monte Carlo approach (Hitchens et al., 2002). In particular, the consequences of missing amide resonances on the reliability of assignments derived from C alpha and CO or from C alpha and C beta chemical shifts was investigated. Missing amide resonances reduce both the number of correct assignments as well as the confidence in these assignments. More significantly, a number of undetectable errors can arise when as few as 9% of the amide resonances are missing from the spectra. However, the use of information from residue specific labeling as well as local and long-range distance constraints improves the reliability and extent of assignment. It is also shown that missing residues have only a minor effect on the assignment of protein-ligand complexes using C alpha and CO chemical shifts and C alpha inter-residue connectivity, provided that the known chemical shifts of the unliganded protein are utilized in the assignment process.
Journal of biomolecular NMR, 25, 1, 11-23.