The nature of CuA in cytochrome c oxidase
Tom H. Stevens, Craig T. Martin, Hsin Wang, Gary W. Brudvig, Charles P. Scholes, Sunney I. Chan, Journal of Biological Chemistry 257, 12106-12113, 1982
The isolation and purification of yeast cytochrome c oxidase is described. Characterization of the purified protein indicates that it is spectroscopically identical with cytochrome c oxidase isolated from beef heart. Preparations of isotopically substituted yeast cytochrome c oxidase are obtained incorporating [1,3-15N2]histidine or [beta,beta-2H2]cysteine. Electron paramagnetic resonance and electron nuclear double resonance spectra of the isotopically substituted proteins identify unambiguously at least 1 cysteine and 1 histidine as ligands to CuA and suggest that substantial spin density is delocalized onto a cysteine sulfur in the oxidized protein to render the site Cu(I)–S.