title: Characterization of the copper(II) binding site in the pink copper binding protein CusF by electron paramagnetic resonance spectroscopy.
abstract: Electron paramagnetic resonance (EPR) spectroscopy has been used to structurally characterize the copper-binding site in CusF protein from Escherichia coli. The EPR spectra indicate a single type II copper center with parameters typical for nitrogen and oxygen ligands (A(parallel) approximately 200 G, g(parallel) approximately 2.186, g(perpendicular) approximately 2.051). The pulsed EPR data show that one of the ligands to Cu2+ is an imidazole ring of a histidine residue. The remote amino nitrogen of this imidazole ring is readily observed by electron spin-echo envelope modulation spectroscopy, while the imino nitrogen that is directly coordinated to the Cu2+ ion is observed by pulsed electron-nuclear double resonance (ENDOR). In addition, the ENDOR spectra reveal the presence of one more nitrogen ligand that was assigned to be a deprotonated peptide nitrogen. Apart from the two nitrogen ligands, it has been established that there are two nearby hydroxyl protons, although whether these belong to a single equatorial water ligand or two equatorial hydroxide ligands is not known.
authors.affiliation: Department of Chemistry, University of Arizona, Tucson, AZ 85721, USA.
authors.forename: Andrei V
authors.initials: AV
authors.lastname: Astashkin
authors.forename: Arnold M
authors.initials: AM
authors.lastname: Raitsimring
authors.forename: F Ann
authors.initials: FA
authors.lastname: Walker
authors.forename: Christopher
authors.initials: C
authors.lastname: Rensing
authors.forename: Megan M
authors.initials: MM
authors.lastname: McEvoy
mesh_terms: D001665:Binding Sites; D002352:Carrier Proteins / Q000737:chemistry* / Q000378:metabolism*; D003300:Copper / Q000378:metabolism*; D004578:Electron Spin Resonance Spectroscopy; D004926:Escherichia coli / Q000737:chemistry*; D006639:Histidine / Q000737:chemistry; D008024:Ligands; D009584:Nitrogen / Q000737:chemistry; D011522:Protons
chemical_list: D002352:Carrier Proteins; D008024:Ligands; D011522:Protons; C023548:copper-binding protein; D006639:Histidine; D003300:Copper; D009584:Nitrogen