Chapter 20

Electron Transport and

Oxidative
Phosphorylation

Paul D. Adams • University of Arkansas

cengage.com/chemistry/campbell
Chapter Outline
20-1 The Role of Electron Transport in Metabolism

20-2 Reduction Potential sin the Electron Transport Chain

20-3 Organization of Electron Transport Complexes

20-4 The Connection between Electron Transport and Phosphorylation

20-5 The Mechanism of Coupling in Oxidative Phosphorylation

20-6 Shuttle Mechanisms

20-7 The ATP Yield from Complete Oxidation of Glucose

The Role of Electron Transport in
Metabolism
• Electron transport is carried out by four closely
related multisubunit membrane-bound complexes
and two electron carriers, coenzyme Q and
cytochrome c
• In a series of oxidation-reduction reactions, electrons
from FADH2 and NADH are transferred from one
complex to the next until they reach O2
• O2 is reduced to H2O
• As a result of electron transport, protons are pumped
across the inner membrane to the intermembrane
space, creating a pH gradient
ATP Production in the Mitochondrion
• The production of ATP in the mitochondria is the
result of oxidative phosphorylation

• The proton gradient establishes a voltage gradient

• The proton and voltage gradients together provide

the mechanism to couple electron transport with
phosphorylation of ADP
Establishment of the Proton Gradient
Summary
• Electron transport from one
carrier to another creates a
proton gradient across the
inner mitochondrial
membrane
• The proton gradient is
coupled to the production of
ATP in aerobic metabolism
Reduction Potentials in the Electron
Transport Chain
• A useful way to look at electron transport is to
consider the change in free energy associated with
the movement of electrons from one carrier to
another
• If we have two electron carriers, for example NADH
and coenzyme Q, are electrons more likely to be
transferred from NADH to coenzyme Q, or vice versa?
• What we need to know is the reduction potential for
each carrier
• A carrier of high reduction potential will tend to be
reduced if it is paired with a carrier of lower reduction
potential
Reduction Potentials (Cont’d)
Reduction Potentials (Cont’d)
Summary
• Standard reduction potentials provide a basis for
comparison among redox reactions

• The sequence of reactions in the electron transport

chain can be predicted by using reduction potentials.
Organization of Electron Transport
Complexes
• Complex I: NADH-CoQ
oxidoreductase
• Electrons are passed from
NADH to FMN

NADH + H+ E-FMN ----> NAD+

+ E-FMNH2
Oxidized and Reduced Forms of CoQ: The
Ubiquinone
Organization of Electron Transport
Complexes (Cont’d)
• Electrons are then passed to the iron-sulfur clusters
• The last step of Complex I involves electrons being
passed to coenzyme Q (also called ubiquinone)
Energetics of Electron Transport
• The transfer of electrons is strongly exergonic and is
sufficient to drive the phosphorylation of ADP
Electron Transport Complexes (Cont’d)
• Complex II: Succinate-coenzyme Q oxidoreductase

Succinate + E-FAD ---> Fumarate + E-FADH2

• The overall reaction is exergonic (-13.5 kj•mol-1), but

not enough to drive ATP production

• No H+ is pumped out of the matrix during this step

Electron Transport Complexes (Cont’d)
• Complex III: CoQH2-cytochrome c oxidoreductase

CoQH2 + 2Cyt c[Fe(III)]--->CoQ + 2Cyt c[Fe(II)] + 2H+

• This reactions of this complex results in a decrease in free

energy that is sufficient to drive the phosphorylation of ADP to
ATP
• The flow of electrons from reduced CoQ, a quinone that can
exist in 3 forms, is known as the Q cycle
Compositions and Locations of Respiratory
Complexes in Inner Mitochondrial Membrane
Oxidized and Reduced Forms of CoQ
Electron Transport Complexes (Cont’d)
• Complex IV: Cytochrome c oxidase
• Catalyzes the final step in electron transport

2Cyt c[Fe(II)] + 2H+ + 1/2O2 ---> 2 Cyt c[Fe(III)] + H2O

• Complex IV contains cytochrome a, cytochrome a3,

and Cu(II), which are also involved in the electron
transport
• Complex IV is the link to molecular oxygen
The Energetics of Electron Transport Reactions
The Heme Group of Cytochromes
• All cytochromes contain a heme group
• There are differences in the side chain depending on
the heme
Fe-S Bonding in Nonheme Iron Proteins
The Connection between Electron
Transport and Phosphorylation
• The energy-releasing oxidations give rise to proton
pumping and a pH gradient across the inner
mitochondrial membrane
• Differences in the concentration of ions across the
membrane generates a voltage gradient
• A coupling process converts the electrochemical
potential to the chemical energy of ATP
• The coupling factor is ATP synthase, a complex
protein oligomer, separate from the electron transport
complexes
• Uncouplers inhibit the phosphorylation of ADP without
affecting electron transport; examples are 2,4-
dinitrophenol, valinomycin, and gramicidin A
F1 and F0 Components of ATP Synthase
Uncouplers
P/O Ratio
• P/O ratio: the number of moles of Pi consumed in
phosphorylation to the number of moles of oxygen
atoms consumed in oxidation
• Phosphorylation: ADP + Pi ----> ATP + H2O
• Oxidation: 1/2O2 + 2H+ + 2e- ---> H2O

• P/O = 2.5 when NADH is oxidized

• P/O = 1.5 when FADH2 is oxidized
Summary
• The coupling of electron transport to oxidative
phosphorylation requires a multisubunit membrane-
bound enzyme, ATP synthase. This enzyme has a
channel for protons to flow from the intermembrane
space into the mitochondrial matrix.

• The proton flow is coupled to ATP production in a

process that appears to involve a conformational
change of the enzyme.
The Mechanism of Coupling in Oxidative
Phosphorylation
• Chemiosmotic coupling
• based on a proton concentration gradient between the
intermembrane space and the matrix
• a proton gradient exists because the various proteins
that serve as electron carriers are not symmetrically
oriented with respect to the two sides of the inner
mitochondrial membrane
• these proteins take up protons from the matrix when
they are reduced and release them to the
intermembrane space when they are reoxidized
• the reactions of NADH, CoQ, and O2 all require
protons
The Mechanism of Coupling in Oxidative
Phosphorylation(Cont’d)
The Mechanism of Coupling in Oxidative
Phosphorylation (Cont’d)
• Evidence for chemiosmotic coupling suggested by Mitchell
(1961):
• A system with definite inside and outside compartments (closed
vesicles) is essential.

• Submitochondrial vesicles can be prepared, which carry out

oxidative phosphorylation and have an asymmetric orientation of
respiratory complexes (Figure 20.16).

• A model system for oxidative phosphorylation can be

constructed with proton pumping in the absence of electron
transport; the model system consists of reconstituted membrane
vesicles, mitochondrial ATP synthase, and a proton pump
(Figure 20.17).

• The existence of the pH gradient has been demonstrated and

confirmed experimentally
The Mechanism of Coupling in Oxidative
Phosphorylation (Cont’d)
• The mechanism by which the proton gradient leads to
the production of ATP depends on ion channels
through the inner mitochondrial membrane
• Protons flow back into the matrix through channels in
the F0 unit of ATP synthase
• The flow of protons is accompanied by formation of
ATP in the F1 unit of ATP synthase
• The details of how phosphorylation takes place as a
result of the linkage to the proton gradient are not
explicitly specified by this mechanism
The Mechanism of Coupling in Oxidative
Phosphorylation (Cont’d)
Conformational Coupling
• The proton gradient leads to changes in conformation
in a number of proteins, including ATP synthase
• There are 3 sites for substrate on ATP synthase, and 3
possible conformations:
- Open (O); a low affinity for substrate
- Loose-binding (L); not catalytically active, binds ADP
and Pi
- Tight-binding (T); catalytically active, binds ATP
• These sites interconvert as a result of proton flux
through ATP synthase
• Proton flux converts L to T, which produces ATP
• Proton flux converts T to O, releasing ATP
Release of ATP from ATP Synthase
Summary
• In chemiosmotic coupling, the proton gradient is the
crux of the matter. The flow of protons through pore
in the synthase drives ATP production.

• In conformational coupling, a change in the shape of

the synthase releases bound ATP that has already
been formed.
Shuttle Mechanisms
• Shuttle mechanisms: transport metabolites between
mitochondria and cytosol
• Glycerol phosphate shuttle:
• We know glycolysis in the cytosol produces NADH
• NADH does not cross the mitochondrial membrane,
but glycerol phosphate and dihydroxyacetone
phosphate do
• Through the glycerol phosphate shuttle, 1.5 ATP are
produced in the mitochondria for each cytosolic NADH
The Glycerol-Phosphate Shuttle
The Malate-Aspartate Shuttle
• The Malate-Aspartate Shuttle:
• Has been found in mammalian kidney, liver, and heart
• Malate crosses the mitochondrial membrane, while
oxaloacetate cannot
• The transfer of electrons from NADH in the cytosol
produces NADH in the mitochondria
• In the malate-aspartate shuttle, 2.5 mitochondrial ATP
are produced for each cytosolic NADH
The Malate-Aspartate Shuttle (Cont’d)
Summary
• Shuttle mechanisms transfer electrons, but not
NADH, from the cytosol across the mitochondrial
membrane

• In the malate-aspartate shuttle, 2.5 molecules of ATP

are produced for each molecule of cytosolic NADH,
rather than 1.5 ATP in the glycerol-phosphate shuttle,
a point that affects the overall yield of ATP in these
tissues
The ATP Yield from Complete Oxidation of
Glucose
• In the complete oxidation of glucose, a total of 30 or
32 molecules of ATP are produced for each
molecule of glucose, depending on the shuttle
mechanism
The ATP Yield from Complete Oxidation of
Glucose (Cont’d)