title: Simultaneous measurement of intra- and intermolecular NOEs in differentially labeled protein-ligand complexes.
abstract: A new NOE strategy is presented that allows the simultaneous observation of intermolecular and intramolecular NOEs between an unlabeled ligand and a 13C,15N-labeled protein. The method uses an adiabatic 13C inversion pulse optimized to an empirically observed relationship between 1 J(CH) and carbon chemical shift to selectively invert the protein protons (attached to 13C). Two NOESY data sets are recorded where the intermolecular and intramolecular NOESY cross peaks have either equal or opposite signs, respectively. Addition and subtraction yield two NOESY spectra which contain either NOEs within the labeled protein (or unlabeled ligand) or along the binding interface. The method is demonstrated with an application to the B12-binding subunit of Glutamate Mutase from Clostridium tetanomorphum complexed with the B12-nucleotide loop moiety of the natural cofactor adenosylcobalamin (Coenzyme B12).
authors.affiliation: Institute of Organic Chemistry, University of Innsbruck, Austria.
authors.forename: C
authors.initials: C
authors.lastname: Eichmüller
authors.forename: W
authors.initials: W
authors.lastname: Schüler
authors.forename: R
authors.initials: R
authors.lastname: Konrat
authors.forename: B
authors.initials: B
authors.lastname: Kräutler
mesh_terms: D001665:Binding Sites; D002247:Carbon Isotopes; D003013:Clostridium / Q000201:enzymology; D003038:Cobamides / Q000737:chemistry; D019751:Intramolecular Transferases / Q000737:chemistry; D008024:Ligands; D046911:Macromolecular Substances; D008958:Models, Molecular; D009587:Nitrogen Isotopes; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D011487:Protein Conformation; D021122:Protein Subunits; D011506:Proteins / Q000737:chemistry*
chemical_list: D002247:Carbon Isotopes; D003038:Cobamides; D008024:Ligands; D046911:Macromolecular Substances; D009587:Nitrogen Isotopes; D021122:Protein Subunits; D011506:Proteins; D019751:Intramolecular Transferases; C021570:methylaspartate mutase; C000913:cobamamide