Role of phospholipid in the low affinity reactions between cytochrome c and cytochrome oxidase

AbstractThe steady-state oxidation of ferrocytochrome c by cytochrome oxidase monitored spectrophotometrically showed that: (1) the kinetics were strictly biphasic with purified enzyme, while mitochondrial membrane-bound enzyme exhibited multiphasic kinetics with extended low affinity phases; (2) the TNmax for the highest affinity phase was as slow as 5–10 electron · s−1 for both preparations, while for the low affinity phases it was about 45 electron · s−1 for the purified enzyme and 150 electron · s−1 for the mitochondrial membrane-bound enzyme; (3) reconstitution of purified enzyme into acidic phospholipid vesicles partially repleted the extended low affinity phases, while reconstitution into uncharged vesicles had no effect