As an organelle, mitochondria is the "power factory" in cells and the main site of aerobic respiration of cells. Its function is to convert energy through oxidative phosphorylation to provide energy for cellular activities. The oxidation process is carried out by four respiratory chain membrane protein complexes (complexes I, II, III and IV) on the inner mitochondrial membrane. mitochondrial complex III, also known as cytochrome c reductase complex, its main function is to oxidize the reduced coenzyme Q10 formed by mitochondrial complexes I and II to oxidative coenzyme Q10. In this process, the OD value increased at 550 nm. Therefore, the activity of mitochondrial complex III can be quantified by measure the change OD value at 550 nm.
Mitochondrial complex I catalyzes the reaction of NADH with ubiquinone substrate to generate NAD+ and reduced ubiquinone. The specific inhibitor rotenone can inhibit the enzyme, and the total enzyme activity minus the enzyme activity after inhibitor inhibition can reflect its specific enzyme activity.
Mitochondrial complex IV can catalyze the oxidation of reduced cytochrome C to oxidized cytochrome C, which has an absorption wavelength at 550 nm. Therefore, the activity of mitochondrial complex IV can be quantified by measure the change OD value at 550 nm.
Mitochondrial complex V is also known as F0F1-ATP synthase. ATP is hydrolyzed by F0F1-ATP synthase to produce ADP, and ADP converts NADH into oxidized coenzyme I (NAD) after enzyme conversion reaction. Therefore, the activity of mitochondrial complex IV can be quantified by measure the change OD value at 340 nm.
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