ATP synthase activity measurement

ATP synthase activity was determined by a commercially available ATP Synthase Enzyme Activity Microplate Assay Kit (Abcam, Cambridge, MA) according to the manufacturer’s instructions. The kit is designed to immunocapture ATP synthase proteins within the wells. The immunocaptured ATP synthase enzyme operates in the “reverse” direction as the physiological role of ATP synthase: it hydrolyzes ATP to ADP and phosphate. This production of ADP is ultimately coupled to the oxidation of NADH to NAD+ which is monitored as a decrease in absorbance at 340 nm. The change in absorbance at 340 nm between 12 and 30 minutes were record. Rate is calculated as ΔOD (m)/ΔT(min).