64] ATP formation in mitochondria, submitochondrial particles, and F0F1 liposomes driven by electric pulses

Abstract

Publisher Summary This chapter describes the procedures for the ATP formation in mitochondria, submitochondrial particles, and F 0 F 1 liposomes driven by electric pulses. The advantage of using electric pulses over artificially imposed ion gradients is that time resolution is excellent and the energy component is simple. To confirm that the observed ATP synthesis takes place on the proton-translocating ATPase (F 0 F 1 ) without participation of other components, F 0 F 1 liposomes were reconstituted and their electric pulse-driven ATP synthesis was measured. Although the pulse-driven ATP synthesis can be observed in F 0 F l microliposomes by applying high-voltage pulses that decay exponentially; rectangular pulses are preferable for quantitative estimation of the applied energy. This method is also applicable to intact mitochondria, submitochondrial particles, and F 0 F 1 liposomes that have large diameters.