Spectrophotometric characterization of intermediate redox states of cytochrome oxidase.

The spectrophotometric characteristics of hemes a and a3 in cytochrome oxidase have been examined over the range 380 nm to 900 nm. Difference spectra (relative to the oxidized form) are presented for ferrous, high-spin oxidized, low-spin oxidized, early "pulsed," late "pulsed," and two-peroxide-treated states of the enzyme. Comparisons indicate that the decay product of the initial peroxide complex of the enzyme is identical to a low-spin pulsed form of the enzyme. A high-spin pulsed form of the enzyme persists for several hours to days after preparation.