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J Am Chem Soc. 2004 Jun 9;126(22):7119-25.

Methyl groups as probes for proteins and complexes in in-cell NMR experiments.

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Graduate Group in Biophysics, University of California San Francisco, San Francisco, California 94143, USA.


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 [(13)C]methyl groups on methionine and alanine provides excellent sensitivity with low background levels at very low costs.

[Indexed for MEDLINE]

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