title: Methyl groups as probes for proteins and complexes in in-cell NMR experiments.
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.
authors.affiliation: Graduate Group in Biophysics, University of California San Francisco, San Francisco, California 94143, USA.
authors.forename: Zach
authors.initials: Z
authors.lastname: Serber
authors.forename: Wesley
authors.initials: W
authors.lastname: Straub
authors.forename: Lorenzo
authors.initials: L
authors.lastname: Corsini
authors.forename: Anson M
authors.initials: AM
authors.lastname: Nomura
authors.forename: Nobuhisa
authors.initials: N
authors.lastname: Shimba
authors.forename: Charles S
authors.initials: CS
authors.lastname: Craik
authors.forename: Paul
authors.initials: P
authors.lastname: Ortiz de Montellano
authors.forename: Volker
authors.initials: V
authors.lastname: Dötsch
mesh_terms: D002247:Carbon Isotopes / Q000737:chemistry; D002477:Cells / Q000737:chemistry*; D004926:Escherichia coli / Q000737:chemistry*; D008715:Methionine / Q000737:chemistry; D008745:Methylation; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D004364:Pharmaceutical Preparations / Q000737:chemistry / Q000378:metabolism; D011506:Proteins / Q000737:chemistry*; D019289:Pyruvic Acid / Q000737:chemistry; D022021:Tacrolimus Binding Proteins / Q000737:chemistry
chemical_list: D002247:Carbon Isotopes; D004364:Pharmaceutical Preparations; D011506:Proteins; D019289:Pyruvic Acid; D008715:Methionine; D022021:Tacrolimus Binding Proteins