ATP oscillation caused by Gradual Entry of Substrates within Mitochondria

The gradual entry of the substrate into enzyme solution causes the oscillatory reaction. We proved this by using dialysis membrane as a means of gradual entry of substrates 7 . It was considered that a suitable combination of permeation rate through membrane of the substrate and rate constants for catalytic reaction regulates the oscillation. It was suggested that such oscillatory reactions also should occur in actual living organisms. On the basis of this suggestion, we explored the oscillatory reaction within mitochondria, because many reactions in mitochondria also occur in the mediation of membrane. We found that the gradual entry of pyruvate together with ADP caused the oscillations of both NADH and ATP within mitochondria. Likewise, the gradual entry of NAD + and malate together with ADP caused the oscillations of NADH and of ATP. At the same time, pH oscillation within mitochondria was also observed. Besides the experiment with mitochondria, we also investigated oscillations of NADH and the other intermediates using dialysis membrane both in the citric acid cycle and in the respiratory chain. Putting these together, we concluded that the oscillatory reactions caused by the gradual entry of pyruvate occur continuously both in the citric acid cycle and in the respiratory chain and is taken over finally by the reaction of ATP synthase in the oxidative phosphorylation, inducing the oscillation of ATP. Furthermore, it was found that oscillations occurred without going through the citric acid cycle when NAD + and malate were used instead of pyruvate.