Spectroscopic studies of ascorbate oxidase. Electronic structure of the blue copper sites.
Low-temperature electronic absorption and room temperature circular dichroism (CD) and magnetic circular dichroism (MCD) spectra are reported for ascorbate oxidase. Bands attributable to d-d electronic transitions in the type 1 (blue) coppers (2B2 ground state) have been observed at 5800, 10000, and 12000 cm-1 (1725, 1000, and 835 nm). The three bands are assigned to the transitions 2B2 leads to 2E, 2B2 leads to 2B1, and 2B2 leads to 2A1, respectively, in the slightly flattened tetrahedral blue copper centers. The ligand field theoretical treatment predicts a type 1 copper reduction potential of 417 mV. The MCD spectrum of ascorbate oxidase contains one major band at 14000 cm-1 (710 nm) and a very weak feature at 13 200 cm-1 (550 nm). The intensities of the CD and MCD bands are consistent with two or three type 1 coppers in each enzyme molecule, and the near-infrared spectroscopic results suggest that the electronic structures of the blue coppers are closely similar.
D001204:Ascorbate Oxidase / Q000378:metabolism*; D001665:Binding Sites; D002942:Circular Dichroism; D003300:Copper / Q000032:analysis*; D010088:Oxidoreductases / Q000378:metabolism*; D011485:Protein Binding; D011487:Protein Conformation; D013053:Spectrophotometry; D013696:Temperature
D003300:Copper; D010088:Oxidoreductases; D001204:Ascorbate Oxidase