20.

6 What Is the P/O Ratio for Mitochondrial Oxidative

Phosphorylation?
The P/O ratio is the number of molecules of ATP formed in oxidative phosphorylation per two electrons
flowing through a defined segment of the electrontransport chain. In spite of intense study of this ratio, its
actual value remains a matter
of contention.
The P/O ratio depends on the ratio of H transported out of the matrix per 2 e
passed from NADH to O2 in the electron-transport chain and on the number of H
that pass through the ATP synthase to synthesize an ATP. The latter number depends
on the number of c-subunits in the F0 ring of the synthase. As noted in Table 20.3, the
number of c-subunits in the ATP synthase ranges from 10 to 15, depending on the organism. This would
correspond to ratios of H consumed per ATP from about 3 to 5,
respectively, since each rotation of the ATP synthase rotor drives the formation of
three ATP. Adding one H for the action of the ATP–ADP translocase raises these values to about 4 and 6,
respectively.
If we accept the value of 10 H transported out of the matrix per 2 e passed from
NADH to O2 through the electron-transport chain, and agree that 4 H are transported into the matrix per
ATP synthesized (and translocated), then the mitochondrial P/O ratio is 10/4, or 2.5, for the case of
electrons entering the electrontransport chain as NADH. This is somewhat lower than earlier estimates,
which placed
the P/O ratio at 3 for mitochondrial oxidation of NADH. For the portion of the chain
from succinate to O2, the H/2e ratio is 6 (as noted previously), and the P/O ratio in
this case would be 6/4, or 1.5; earlier estimates placed this number at 2. The consensus of more recent
experimental measurements of P/O ratios for these two cases has
been closer to the values of 2.5 and 1.5. Many chemists and biochemists, accustomed
to the integral stoichiometries of chemical and metabolic reactions, were once reluctant to accept the
notion of nonintegral P/O ratios. At some point, as we learn more
about these complex coupled processes, it may be necessary to reassess the numbers.

20.7 How Are the Electrons of Cytosolic NADH Fed

into Electron Transport?
Most of the NADH used in electron transport is produced in the mitochondrial
matrix, an appropriate site because NADH is oxidized by Complex I on the matrix side of the inner
membrane. Furthermore, the inner mitochondrial membrane is impermeable to NADH. Recall, however,
that NADH is produced in
glycolysis by glyceraldehyde-3-P dehydrogenase in the cytosol. If this NADH
were not oxidized to regenerate NAD, the glycolytic pathway would cease to
function due to NAD limitation. Eukaryotic cells have a number of shuttle
systems that harvest the electrons of cytosolic NADH for delivery to mitochon-