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Methyl groups as probes for proteins and complexes in in-cell NMR experiments
Authors:Serber Zach  Straub Wesley  Corsini Lorenzo  Nomura Anson M  Shimba Nobuhisa  Craik Charles S  Ortiz de Montellano Paul  Dötsch Volker
Affiliation:Graduate Group in Biophysics, University of California San Francisco, San Francisco, California 94143, USA.
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 [(13)C]methyl groups on methionine and alanine provides excellent sensitivity with low background levels at very low costs.
Keywords:
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