Dicyclohexylcarbodiimide Does not Inhibit Proton Pumping by Cytochrome c Oxidase of Paracoccus denitrificans

Abstract

The effect of dicyclohexylcarbodiimide (DCCD) on the proton pumping 2 subunit cytochrome c oxidase from P. denitrificans was investigated. Purified Paracoccus oxidase was reconstituted into phospholipid vesicles by cholate dialysis. Following incubation with increasing amounts of DCCD, proton ejection was recorded in response to reductant pulses with reduced cytochrome c. Concentrations of DCCD which greatly reduced proton pumping by bovine cytochrome c oxidase used as a control exert only a minor effect on proton translocation by Paracoccus oxidase. Incubation of the bacterial enzyme with [14C]DCCD failed to reveal the specific covalent interaction previously demonstrated to occur with bovine cytochrome c oxidase and is also shown for the oxidase of yeast. Paracoccus oxidase differs in its interaction with DCCD from the functionally analogous eukaryotic enzymes.