Effects of uncouplers

and inhibitors of the

electron transport-
oxidative
phosphorylation
system
 The coupling between electron
transport and ATP synthesis can be
abolished by chemical reagents, or
uncouplers, such as 2,4- Dinitrophenol
or carbonyl cyanide-p-
trifluoromethoxyphenylhydrazone.
 After addition of an uncoupler to a
suspension of tightly coupled liver
mitochondria with a low rate of O2
uptake, a rapid increase in the rate of
O2 consumption is observed(fig.14.43a).
 Because electron transport is uncoupled
from ATP synthesis, electron transport
may continue but without ATP synthesis.
 Uncouplers are hydrophobic weak acids
that pick up a proton in the
intermembrane space where a higher
concentration of protons results from
active electron transfer.

 These protonated uncouplers, being

lipophilic, rapidly diffuse into the
mitochondrial matrix where they lose
their proton, because of the lower
concentration of the protons there.
 In this way, the proton gradient can be
completely dissipated as shown in fig
14.44 and hence ATP synthesis is
abolished.
 As illustrated in fig 14.43b, addition of oligomycin, and inhibitor of the ATP synthase, 2
liver mitochondria actively respiring in the presence ADP results in an inhibition of O2
uptake.
 Oligomycin blocks the synthesis of ATP by preventing the movement of protons through
the ATP synthase.
 Since, ATP synthesis and electron flow are tightly coupled, the build up of protons in
the intermembrane space almost completely blocks the rate of electron transport for
respiratory control.
 Subsequent addition of 2,4- Dinitrophenol that dissipates the proton gradient results in
a rapid increase in the rate of O2 uptake, because electron transport has been
uncoupled from ATP synthesis.
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