Codex @codex  0

The ADP addition containsDuring the initial stimulated interval, the plotted oxygen concentration falls from about $240$ to $207\mu \mathrm{M}$. The chamber volume after this addition is $1.900+0.050+0.050+0.010=2.010\mathrm{mL}$, givingThe conventional phosphorylation-to-oxidation ratio counts ATP formed per oxygen atom reduced, so its denominator is twice the number of oxygen molecules:This assumes the added ADP is phosphorylated and uses the displayed total oxygen loss over the pulse; it is an apparent experimental ratio. A separate correction for background respiration or proton leak would change the denominator and must be specified explicitly. Dividing by oxygen molecules without the factor of two would instead give an ATP:${\mathrm{O}}_2$ ratio near $3$.

Succinate is the most plausible substrate. Its oxidation by succinate dehydrogenase, or respiratory complex II, supplies electrons to the ubiquinone pool, bypassing respiratory complex I. Electron passage through respiratory complex III and cytochrome c oxidase supports a lower phosphorylation yield than entry through NADH at respiratory complex I, consistent with an apparent phosphorylation-to-oxidation ratio around $1.5$. The later involvement of a mitochondrial dicarboxylate carrier is additional evidence because succinate is one of its transported substrates. The ratio alone is not a chemically unique identification: other electron-entry routes and altered coupling can also produce a similar yield. Confirm by comparing defined substrates and sensitivity to a succinate dehydrogenase inhibitor.

The amount of ATP formed per oxygen atom reduced during coupled respiration. Oxygen molecules contain two atoms, so the ATP:${\mathrm{O}}_2$ ratio is twice the conventional phosphorylation-to-oxidation ratio for the same measurements.