title: The ligand geometry of copper determines the stability of amicyanin.
abstract: Solution differential scanning calorimetry (DSC) of oxidized amicyanin, a Type I copper protein, at pH 7.5 reveals two thermal transitions. The major transition at 67.7 degrees C corresponds to the disruption of the Cys(92) thiolate to Cu(II) charge transfer as evidenced by a corresponding temperature-dependent loss of amicyanin visible absorbance. A minor transition at 75.5 degrees C describes the further irreversible protein unfolding. Reduced amicyanin exhibits a pH-dependent change of the copper ligand geometry. At pH 8.5 where the Type I tetrahedral geometry is maintained, DSC reveals two thermal transitions with T(m) values similar to that of oxidized amicyanin. At pH 6.2 where the Cu(I) coordination is trigonal planar, reduced amicyanin exhibits a single thermal transition with a lower T(m) of 64.0 degrees C. Apoamicyanin, from which copper has been removed, also exhibits a single thermal transition but with a much lower T(m) of 51.8 degrees C. Thus, the thermal stability of amicyanin is dictated both by the presence or absence of copper and its ligand geometry, but not its redox state. The physiological relevance of these data is discussed.
authors.affiliation: Department of Biochemistry, The University of Mississippi Medical Center, Jackson, MS 39216-4505, USA.
authors.forename: John K
authors.initials: JK
authors.lastname: Ma
authors.forename: G Reid
authors.initials: GR
authors.lastname: Bishop
authors.forename: Victor L
authors.initials: VL
authors.lastname: Davidson
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry* / Q000302:isolation & purification; D002152:Calorimetry, Differential Scanning; D003300:Copper / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D008024:Ligands; D008958:Models, Molecular; D010084:Oxidation-Reduction; D010231:Paracoccus denitrificans / Q000737:chemistry*; D011487:Protein Conformation; D013053:Spectrophotometry; D013816:Thermodynamics
chemical_list: D001426:Bacterial Proteins; D008024:Ligands; D003300:Copper