Biochemical pathway for the biosynthesis of the Cu_A center in bacterial cytochrome c oxidase

The di-copper center Cu_A is an essential metal cofactor in cytochrome oxidase (Cox) of mitochondria and many prokaryotes, mediating one-electron transfer from cytochrome c to the site for oxygen reduction. Cu_A is located in subunit II (CoxB) of Cox and protrudes into the periplasm of Gram-negative bacteria or the mitochondrial intermembrane space. How the two copper ions are brought together to build CoxB·Cu_A is the subject of this review. It had been known that the reductase TlpA and the metallochaperones ScoI and PcuC are required for Cu_A formation in bacteria, but the mechanism of copper transfer has emerged only recently for the Bradyrhizobium diazoefficiens system. It consists of the following steps: (a) TlpA keeps the active site cysteine pair of CoxB in its dithiol state as a prerequisite for metal insertion; (b) ScoI·Cu²⁺ rapidly forms a transient complex with apo-CoxB; (c) PcuC, loaded with Cu¹⁺ and Cu²⁺ , dissociates this complex to CoxB·Cu²⁺ , and a second PcuC·Cu¹⁺ ·Cu²⁺ transfers Cu¹⁺ to CoxB·Cu²⁺ , yielding mature CoxB·Cu_A . Variants of this pathway might exist in other bacteria or mitochondria.