Peculiarities of cyanide binding to the ba3-type cytochrome oxidase from the thermophilic bacterium Thermus thermophilus.

Cytochrome c oxidase of the ba(3)-type from Thermus thermophilus does not interact with cyanide in the oxidized state and acquires the ability to bind heme iron ligands only upon reduction. Cyanide complexes of the reduced heme a(3) in cytochrome ba(3) and in mitochondrial aa(3)-type cytochrome oxidase are similar spectroscopically, but the a(3)(2+)-CN complex of cytochrome ba(3) is strikingly tight. Experiments have shown that the K(d) value of the cytochrome ba(3) complex with cyanide in the presence of reductants of the enzyme binuclear center does not exceed 10(-8) M, which is four to five orders of magnitude less than the K(d) of the cyanide complex of the reduced heme a(3) of mitochondrial cytochrome oxidase. The tightness of the cytochrome ba(3) complex with cyanide is mainly associated with an extremely slow rate of the ligand dissociation (k(off) < or = 10(-7) sec(-1)), while the rate of binding (k(on) ~ 10(2) M(-1).sec(-1)) is similar to the rate observed for the mitochondrial cytochrome oxidase. It is proposed that cyanide dissociation from the cytochrome ba(3) binuclear center might be hindered sterically by the presence of the second ligand molecule in the coordination sphere of Cu(B)(2+). The rate of cyanide binding with the reduced heme a(3) does not depend on pH in the neutral area, but it approaches linear dependence on H+ activity in the alkaline region. Cyanide binding appears to be controlled by protonation of an enzyme group with pK(a) = 8.75.