Bovine heart cytochrome c oxidase preparations contain high affinity binding sites for magnesium as well as for zinc, copper, and heme iron.

Metal contents of six bovine heart cytochrome c oxidase preparations were determined by inductively coupled plasma atomic emission spectrometry (ICP-AES). Magnesium is present in consistent amounts reflected by Fe/Mg and Mg/Zn ratios of 2.06 +/- 0.18 and 1.10 +/- 0.14, respectively. More copper than iron is always present; the average Cu/Fe atom ratio is 1.27 +/- 0.10. Calcium is found in significant but variable amounts. Mg, Zn, Fe, and Cu are each bound with high affinity as shown by a resistance to removal upon dialysis against various media. The Cu in excess of Fe, the Mg, and the Zn may each have catalytic and/or structural roles in the oxidase. The observed metal stoichiometry suggests that a dimeric catalytic unit with 5 Cu, 4 Fe, 2 Zn, and 2 Mg may be present in the inner mitochondrial membrane to carry out O2 reduction and H+ pumping.