title: Resonance Raman spectroscopy of amicyanin, a blue copper protein from Paracoccus denitrificans.
abstract: The copper binding site of amicyanin from Paracoccus denitrificans has been examined by resonance Raman spectroscopy. The pattern of vibrational modes is clearly similar to those of the blue copper proteins azurin and plastocyanin. Intense resonance-enhanced peaks are observed at 377, 392, and 430 cm-1 as well as weaker overtones and combination bands in the high frequency region. Most of the peaks below 500 cm-1 shift 0.5-1.5 cm-1 to lower energy when the protein is exposed to D2O. Based on the pattern of conserved amino acids, the axial type EPR spectrum, and the resonance Raman spectrum, it is proposed that the copper binding site in amicyanin contains a Cu(II) ion in a distorted trigonal planar geometry with one cysteine and two histidine ligands and an axial methionine ligand at a considerably longer distance. Furthermore, the presence of multiple intense Raman peaks in the 400 cm-1 region which are sensitive to deuterium substitution leads to the conclusion that the Cu-S stretch is coupled with internal ligand vibrational modes and that the sulfur of the cysteine ligand is likely to be hydrogen-bonded to the polypeptide backbone.
authors.affiliation: Department of Chemical and Biological Sciences, Oregon Graduate Center, Beaverton 97006-1999.
authors.forename: K D
authors.initials: KD
authors.lastname: Sharma
authors.forename: T M
authors.initials: TM
authors.lastname: Loehr
authors.forename: J
authors.initials: J
authors.lastname: Sanders-Loehr
authors.forename: M
authors.initials: M
authors.lastname: Husain
authors.forename: V L
authors.initials: VL
authors.lastname: Davidson
mesh_terms: D001400:Azurin; D001426:Bacterial Proteins; D001665:Binding Sites; D003300:Copper / Q000378:metabolism; D003545:Cysteine / Q000378:metabolism; D003903:Deuterium; D004578:Electron Spin Resonance Spectroscopy; D008667:Metalloproteins / Q000378:metabolism; D010231:Paracoccus denitrificans / Q000032:analysis*; D010970:Plastocyanin; D013059:Spectrum Analysis, Raman
chemical_list: D001426:Bacterial Proteins; D008667:Metalloproteins; C017079:mauC protein, Methylobacterium extorquens; D001400:Azurin; D003300:Copper; D010970:Plastocyanin; D003903:Deuterium; D003545:Cysteine