V. Zickermann et al., TRANSFORMATION OF THE CU-A REDOX SITE IN CYTOCHROME-C-OXIDASE INTO A MONONUCLEAR COPPER CENTER, Biochemistry, 36(11), 1997, pp. 3232-3236
Subunit II Of the aa(3) type cytochrome c oxidase contains a binuclear
copper center (CUA) which functions as the entry point for electrons
donated by cytochrome c. We have introduced site-specific mutations in
residues liganding the Cu-A center in the oxidase of the bacterium Pa
racoccus denitrificans; the purified, fully assembled enzyme complexes
were analyzed by various techniques, including EPR, optical spectrosc
opy, and total-reflection X-ray fluorescence spectrometry, to determin
e metal to protein ratios. In the C216S mutant, the binuclear Cu-A Sit
e is transformed into a mononuclear copper center. In contrast to wild
type, the C216S mutant does no longer exhibit the characteristic abso
rption band in the near-infrared region of the optical spectrum that h
as been assigned to Cu-A. These major changes in the CUA Site Of this
mutant correlate with an almost complete loss in catalytic activity.