Fluorescein mercuric acetate specifically displaces zinc from cytochrome oxidase

Treatment of beef heart cytochrome oxidase with fluorescein mercuric acetate (FMA) was found to specifically displace zinc from the enzyme and inhibit the steady-state activity in a parallel fashion. The native cytochrome oxidase preparation contained 2.3 Cu: 2.0 Fe: 1.1 Zn: 0.9 Mg. Addition of 2 equivalents of FMA inhibited the activity by 50% and displaced 60% of the zinc from the enzyme, but did not affect the copper, iron or magnesium content. The pre-steady-state reduction of cytochrome oxidase by ferrocytochrome c was not affected by the FMA treatment, in contrast to the inhibition of steady state activity. These results suggest a possible structural or functional role for zinc in cytochrome oxidase.