Cholate Disrupts Regulatory Functions of Cytochrome c Oxidase

Cytochrome c oxidase (CytOx), the oxygen-accepting and rate-limiting enzyme of mitochondrial respiration, binds with 10 molecules of ADP, 7 of which are exchanged by ATP at high ATP/ADP-ratios. These bound ATP and ADP can be exchanged by cholate, which is generally used for the purification of CytOx. Many crystal structures of isolated CytOx were performed with the enzyme isolated from mitochondria using sodium cholate as a detergent. Cholate, however, dimerizes the enzyme isolated in non-ionic detergents and induces a structural change as evident from a spectral change. Consequently, it turns off the allosteric ATP-inhibition of CytOx, which is reversibly switched on under relaxed conditions via cAMP-dependent phosphorylation and keeps the membrane potential and ROS formation in mitochondria at low levels. This cholate effect gives an insight into the structural-functional relationship of the enzyme with respect to ATP inhibition and its role in mitochondrial respiration and energy production.

Automated summary

Cytochrome c oxidase (CytOx), the key enzyme involved in mitochondrial respiration, binds with ADP and ATP, which can be exchanged by cholate. The use of sodium cholate detergent in isolating CytOx induces structural changes and turns off the allosteric ATP-inhibition, revealing insights into the enzyme's role in energy production.