Electron Transport and

Oxidative Phosphorylation
 An Overview
Biological oxidations are catalyzed by intracellular
enzymes. The purpose of oxidation is to obtain energy.
Electron Transport: Electrons carried by reduced
coenzymes (NADH or FADH2) are passed sequentially
through a chain of proteins and coenzymes (so called
electron transport chain)to O2 .
Oxidative Phosphorylation: Coupling e- Transport
(Oxidation) and ATP synthesis (Phosphorylation) .
It all happens in mitochondrion or at the inner
mitochondrial membrane (eukaryotic cells)
 mitochondrion
the mitochondrion contained the enzymes responsible
for electron transport and oxidative phosphorylation

In inner
membrane
knobs
Impermeable to
ions and most
other compounds
MITOCHONDRION
 Reduction Potentials
E0=standard reduction potential.
The relative tendency to accept e-s and become
Crucial
reduced. equation:
Go' = -nF Eo'

Number of electrons = Eo'(acceptor) - Eo'(donor)

transferred in the Faradays constant
redox reaction (96485 J/volt/mole)

If Eo' is positive, an electron transfer reaction is

spontaneous (Go' <0)
Fumarate+2H++2e- succinate 0.031
FAD+ 2H++2e- FADH2 0
NAD++ 2H++2e- NADH+H+ -0.32

Succinate+FAD Fumarate+ FADH2

G' = - n F E'
Eo' = Eo'(acceptor) - Eo'(donor)
G =-2*96485*(0-0.031)= 5.98KJ/mol

Succinate+ NAD+ Fumarate+ NADH+H+

G =-2*96485*(-0.32- 0.031)= 67.7KJ/mol
 Removal of H across a C-C bond is not
sufficiently exergonic to reduce
NAD+,but it does yield enough energy
to reduce FAD.
Thats why succinate dehydrogenase
uses FAD other than NAD+ as coenzyme.
 Electron Carriers
The transfer of electrons is not directly to oxygen but
through coenzymes
NAD+ There are 2 sites of entry
for electrons into the
FMN electron transport chain:
NAD+ or FAD
FeS
Both are coenzymes for
FAD FeS ubiquinone dehydrogenase enzymes
Cyt b

ubiquinone

FeS Cyt c1 Cyt c Cyt a Cyt a3

1/2 O2
Nicotinamide coenzymes: NAD+

X H H
CONH2 XH2 CONH2

+ H+
N N

oxidised coenzyme reduced coenzyme

+
NAD or NADP
+ NADH or NADPH

Always a 2-electron reaction transferring 2 e- and 2 H+

The flavin coenzymes / flavoproteins

OH OH OH O
CH2 CH CH CH CH2 O P OH
H3C N N O OH

riboflavin monophosphate
N (flavin mononucleotide, FMN)
H3C N

NH2

N
N
OH OH OH O O
CH2 CH CH CH CH2 O P O P O CH2 O N N
H3C N N O OH OH

flavin adenine dinucleotide (FAD)

N
H3C N OH OH
O

FAD Always a 2-electron reaction transferring 2 e- and 2 H+

Oxidation and reduction of flavin coenzymes
O O O
H H H H H
C N C C N C C N C
H3C C C C NH H 3C C C C NH H3C C C C NH

H3C C C C C O H 3C C C C C O H3C C C C C O
C N N C N N C N N
H H H H
CH2 CH2 CH2
- + - +
e +H e +H
HC OH HC OH HC OH

HC OH HC OH HC OH

HC OH O HC OH O HC OH O

H2C O P O- H2C O P O- H2C O P O-

O- O- O-
FMN FMNH FMNH2

it can accept/donate 1 or 2 e-. FMN has an important role in

mediating e- transfer between carriers that transfer 2 e- (e.g.,
NADH) and those that transfer 1 e- (e.g., Fe+++).
Role of FMN mediating between 2e- & 1e-
carriers:
For example, when NADH donates
electrons to the respiratory chain, the
initial electron transfers are:
NADH + H+ + FMN NAD+ + FMNH2
FMNH2 + Fe+++ FMNH + Fe++ + H+
 Iron-sulfur Centers (clusters)

Iron-sulfur centers (Fe-S) are prosthetic groups containing 1-

4 iron atoms
Iron-sulfur centers transfer only one electron, even if
they contain two or more iron atoms.
E.g., a 4-Fe center might cycle between redox states:
Fe+++3, Fe++1 (oxidized) + 1 e- Fe+++2, Fe++2 (reduced)
 Cys S
S Fe
Cys
S Fe S
S Fe S
Cys Cys
S Fe
S

Cys S S S Cys
Fe Fe

Cys S S S Cys

Iron-Sulfur Centers
 NAD+

Ubiquinone FMN
Other names and abbreviations:
FeS
Coenzyme Q CoQ Q
O FAD FeS ubiquinone
CH3O CH3
Cyt b

CH3
ubiquinone
CH3O (CH2 CH C CH2)nH
O FeS
coenzyme Q
2 e- + 2 H+

OH Most often n = 10
CH3O CH3 Free CoQ can undergo a 2 e-
oxidation/reduction:
CH3 Q + 2 e- + 2 H+ QH2.
CH3O (CH2 CH C CH2)nH
OH
coenzyme QH2
When bound to special sites in respiratory complexes, CoQ
can accept 1 e- to form a semiquinone radical (Q-).

O O-
CH3O CH 3 CH3O CH 3

CH 3
e-
CH 3
CH3O (CH 2 CH C CH 2)nH CH3O (CH 2 CH C CH 2)nH
O
coenzyme Q O
coenzyme Q -
e- + 2 H+
OH
CH3O CH 3

CH 3
CH3O (CH 2 CH C CH 2)nH
OH coenzyme QH2
 Coenzyme Q (CoQ, Q or ubiquinone) is
lipid-soluble. It dissolves in the
hydrocarbon core of a membrane.
the only electron carrier not bound to a
protein.
it can accept/donate 1 or 2 e-. Q can mediate
e- transfer between 2 e- that transfer and 1 e-
carriers
Cytochromes

NAD+

FMN proteins that accept

FeS
electrons from QH2 or
FeS
FAD FeS ubiquinone

Ultimately transfers the

Cyt b
electrons to oxygen
ubiquinone

FeS Cyt c1 Cyt c Cyt a Cyt a3

1/2 O2
 Cytochromes
Cytochromes are electron carriers
containing hemes . Hemes in the 3 classes
of cytochrome (a, b, c) differ in substituents
on the porphyrin ring.
Some cytochromes(b,c1,a,a3) are part of
large integral membrane protein
complexes.
Cytochrome c is a small, water-soluble
protein.
 CH3

CH3 HC S CH2 protein

N
H3C CH3
N Fe N
- protein
OOC CH2 CH2 CH S CH2
N
CH3

CH2 CH3

CH2

COO- Heme c
Heme is a prosthetic group of cytochromes.
Heme contains an iron atom in a porphyrin ring
system.
The heme iron can undergo 1 e- transition
between ferric and ferrous states: Fe3+ + e-
Fe2+
Copper ions besides two heme A groups (a
and a3) act as electron carriers in Cyta,a3
Cu2++e- Cu+
 Electron carriers
NAD+, flavins and Q carry electrons and H+
Cytochromes and non-haem iron proteins carry only
electrons

NAD+ FAD undergoes only a 2 e- reaction;

cytochromes undergo only 1e- reactions
FMN Q undergoes 1e- and 2 e- reaction
 Electron Transport chain
(respiratory chain)
The electron transport chain in the inner
mitochondrial membrane can be isolated in
four proteins complexes(I, II, III, IV).
A lipid soluble coenzyme (Q) and a water
soluble protein (cyt c) shuttle between
protein complexes
Electrons transfer through the chain - from
complexes I and II to complex IV
The electron transport chain
Mitochondrial Complexes
 Mitochondrial Complexes
NAD+

FMN
I NADH Dehydrogenase
FeS

FAD FeS ubiquinone

II
Cyt b
Succinate
dehydrogenase Cytochrome Oxidase
ubiquinone

FeS Cyt c1 Cyt c Cyt a Cyt a3

III IV
1/2 O2
CoQ-cyt c Reductase
 Support for this order of events
1. Energetically favorable. electrons pass from
lower to higher standard reduction potentials
.2. Spectra: the absorption spectrum for the
reduced carrier differs from that of its oxidized
form.
carriers closer to oxygen are more oxidized.
3. Specific inhibitors. Those before the blocked step
should be reduced and those after be oxidized.
4. Assay of individual complexes.
NADH can reduce complex I but not the other complexes.
Order and Reduction Potentials
NAD+ -0.32

FMN -0.3

FeS

FAD FeS ubiquinone +0.045

+0.03 Cyt b +0.077

ubiquinone +0. 29 +0. 55

FeS Cyt c1 Cyt c Cyt a Cyt a3

+0. 22 +0. 25
1/2 O2

+0.82
Drugs that inhibit the ETC
NAD+

Amytal FMN Rotenone helps natives of the

I Amazon rain forest catch fish!
rotenone
FeS

FAD FeS ubiquinone

II CN- CO
Cyt b
Antimycin A binding tightly to the ferric form
ubiquinone (Fe3+) of a3

FeS Cyt c1 Cyt c Cyt a Cyt a3

III IV
1/2 O2
 When the chain is blocked, electron
carriers will be in a reduced state before
the block point and in an oxidized state
after it.
This can easily be monitored using
difference spectra.
 Inhibitors and Artificial
Electron Acceptors
Rotenone
amytal Antimycin A CN-,CO

Methylene
blue +0.01
Ferricyanide+0.36
H + Transport
Complex I, III, IV drive H+
transport from matrix to the cytosol
When e- flow through, which creates
proton gradient(electrochemical potential)
across the inner membrane
Complex I and Complex IV : The
mechanism of H+ transport is still not
known.
The mechanism of H+ transport in
Complex III is Q cycle.
 Matrix
H+ + NADH NAD+ + 2H+ 2H+ + O2 H2O

2 e-
I Q III IV

++
cyt c
4H+ 4H+ 2H+
Intermembrane Space

4H+ are pumped per 2e- passing through complex III.

The H+/e- ratio is less certain for the other complexes:
probably 4H+/2e- for complex I; 2H+/2e- for complex IV.
 Q Cycle :The mechanism of H+ transport in Complex III

matrix 2 H+
Q Q.- QH2 QH2

cyt bH
Complex III
cyt bL QH2
e- - e-
Q Q. Fe-S cyt c1
intermembrane 2 H+ cyt c
space
1.Electrons are transported along the inner mitochondrial
membrane, through a series of electron carriers
2.Protons (indicated by + charge) are translocated across the
membrane, from the matrix to the intermembrane space
3.Oxygen is the terminal electron acceptor, combining
with electrons and H+ ions to produce water
4. As NADH delivers more H+ and electrons into the ETS,
the proton gradient increases, with H+ building up outside
the inner mitochondrial membrane, and OH- inside the
membrane.