Methyl Groups as Probes for Proteins and Complexes in In-Cell NMR Experiments
- 8 May 2004
- journal article
- research article
- Published by American Chemical Society (ACS) in Journal of the American Chemical Society
- Vol. 126 (22) , 7119-7125
- https://doi.org/10.1021/ja049977k
Abstract
Studying protein components of large intracellular complexes by in-cell NMR has so far been impossible because the backbone resonances are unobservable due to their slow tumbling rates. We describe a methodology that overcomes this difficulty through selective labeling of methyl groups, which possess more favorable relaxation behavior. Comparison of different in-cell labeling schemes with three different proteins, calmodulin, NmerA, and FKBP, shows that selective labeling with [13C]methyl groups on methionine and alanine provides excellent sensitivity with low background levels at very low costs.Keywords
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