Summary of known ETS and Oxidative

Phosphorylation inhibitors
Electron Carrier Inhibitors
Electron Transport Inhibitors
Structures of a Few Inhibitors
 INHIBITORS
• Rotenone is a common insecticide that strongly inhibits the electron
transport of complex I. Rotenone is a natural product obtained from the
roots of several species of plants. Tribes in certain parts of the world beat
the roots of trees along riverbanks to release rotenone into the water
which paralyzes fish and makes them easy prey.
• Amytal is a barbiturate that inhibits the electron transport of complex I.
• Demerol is painkiller that also inhibits complex I.
• All three are complex I inhibitors & block the oxidation of the Fe-S clusters
of complex I.
• 2-Thenoyltrifluoroacetone and carboxin specifically block electron
transport in Complex II.
• Antimycin A1 is an antibiotic that inhibits electron transfer in
complex III by blocking the transfer of electrons between Cyt bH
and coenzyme Q bound at the QN site
• Cyanide, azide and carbon monoxide all inhibit electron transport
in Complex IV. The all inhibit electron transfer by binding tightly
with the iron coordinated in Cyt a3.
• Axide and cyanide bind to the iron when the iron is in the ferric
state.
• Carbon Monoxide binds to the iron when it is in the ferrous state.
• Cyanide and azide are potent inhibitors at this site which accounts
for there acute toxicity.
Carbon monoxide is toxic due to its affinity for the heme iron of hemoglobin. Animals
carry many molecules of hemoglobin, therefore it takes a large quantity of carbon
monoxide to die from carbon monoxide poisoning. Animals have relatively few
molecules of Cyt a3. Consequently an exposure to a small quantity of azide or cyanide
can be lethal. The toxicity of cyanide is solely from its ability to arrest electron
transport.
 Uncouplers

• Uncouplers uncouple electron

transport and oxidative
phosphorylation
• Uncouplers disrupt the tight
coupling between electron
transport and oxidative
phosphorylation by dissipating the
proton gradient
• Uncouplers are hydrophobic
molecules with a dissociable proton
• They shuttle back and forth across
the membrane, carrying protons to
dissipate the gradient
Uncoupling
• H+ gradient dissipated by
leakage of H+ across inner
membrane, bypassing synthase
• result: phosphorylation ,
oxidation 
• uncouplers
• ionophores: small mobile car-
riers of ions across membranes
e.g., dinitrophenol: H+ ionophore
• thermogenin: transmembrane
protein in brown adipose tissue
function: to generate heat
• other mitochondrial proteins:
variants may account for
differences in weight gain/loss

Lehninger et al., Fig 19-28

 Dinitrophenol (DNP) dissipates the proton gradient
by carrying H+ across the inner mitochondrial membrane through
simple diffusion-mediated transport

The result is that

carbohydrate
and lipid stores
are depleted in
an attempt to
make up for the
low energy
charge in cells
resulting from
decreased ATP
synthesis; DNP
short-circuits the
proton circuit.
The UCP1 uncoupling protein, also called thermogenin, controls
thermogenesis in newborn and hibernating animals

Thermogenin is a endogenous
uncoupler present in Brown
fat(brown due high content of mito)
uncouples OP & generates HEAT,thus
maint body temp in new born
&hibernating animals
 Electron transfer to O2 is tightly coupled to ATP synthesis in mitochondria, as is
demonstrated in these experiments

Mitochondria are suspended in a buffered

medium, and an O2 electrode is used to
monitor O2 consumption. At intervals,
samples are removed and assayed for the
presence of ATP. (a)
i) The addition of ADP and Pi alone results
in little or no increase in either respiration
(O2 consumption; black) or ATP synthesis
(red).
ii) When succinate is added, respiration
begins immediately and ATP is synthesized.
iii) The addition of cyanide (CN- ), which
blocks electron transfer between
cytochrome oxidase and O2, inhibits both
respiration and ATP synthesis.
ATP Formation

When isolated mitochondria are suspended in a buffer containing ADP, Pi, and an
oxidizable substrate such as succinate, three easily measured processes occur: (1) the
substrate is oxidized (succinate yields fumarate), (2) O2 is consumed (respiration
occurs), and (3) ATP is synthesized.

Careful experimental measurements of the stoichiometry of electron transfer to O2 and

the associated synthesis of ATP show that with NADH as electron donor, mitochondria
synthesize nearly 3.0 ATP per pair of electrons passed to O2, and with succinate nearly
2.0 ATP per electron pair. Oxygen consumption and ATP synthesis are dependent upon
substrate oxidation, as can be seen in the experiments
 Role of Oligomycin & DNP

(b) Mitochondria provided with

succinate respire and synthesize ATP
only when ADP and Pi are added.
i)Subsequent addition of
venturicidin or oligomycin,
inhibitors of ATP synthase, blocks
both ATP synthesis and respiration.
ii)Dinitrophenol (DNP) allows
respiration to continue without ATP
synthesis; DNP acts as an uncoupler.