LECTURE NOTES ON :

BIOENERGETICS, ELECTRON
TRANSPORT CHAIN &
OXIDATIVE
PHOSPHORYLATION

By

Ajilore B.S. (MBChB, PhD)

 Energetics of Life
 To stay alive, man requires energy

 This energy is derived not just from carbohydrates

but also from other important dietary substances like
◦ lipids,
◦ proteins,
◦ vitamins
◦ Nucleotides etc

 Lifeenergetics involves how this energy is harvested

and utilized from the chemical bonds present in food
substances, through a number of metabolic
pathways

 One important way by which the body harvests

 ATP is a chemical carrier of energy, from which energy can
be obtained and used directly for the performance of
biological activities
ATP
 Is the energy carrier for most biological
reactions;
 It is referred to as the energy currency of cells;
 It is constantly being used to drive endergonic
processes;
 It is highly unstable & spontaneously hydrolyzed
by water to ADP and Pi;
 But in cells, the conc. of ATP is 100X greater
than ADP.
 This means that cells keep the ATP hydrolysis
reaction far displaced (away) from chemical
equilibrium
 This is accompanied by a process known as
cellular energy metabolism
.

The product of this hydrolysis is ADP (adenosine diphosphate) and

phosphate, and is accompanied by release of energy.

5
Introduction to Metabolism

Metabolism is the sum total of all

chemical reactions in living cells;

Itis the overall process through

which living systems acquire
and utilize free energy to
carry out their functions
What is free energy?

 Itis the potential energy of a substance;

 Potential energy is the energy that is not
actively engaged in doing work but has the
potential to do so;
 Free energy is represented by Gibbs (G) & it is
difficult to measure directly
 In any chemical reaction the free energy
content of reactant & product are not the same;
 Hence, the free energy change ∆G when a
substance A is converted to B is given as:

∆G = GB – GA for A → B
When ∆G is -Ve:

Free energy of product is less than

free energy of reactant;
The conversion of A to B is
accompanied by release of free
energy & reaction results in decrease
in free energy & increase in entropy;
The reaction is said to be exergonic &
takes place spontaneously;
 The reaction generates energy in
biological systems
When ∆G is +Ve

Free energy of product is higher than

free energy of reactant
Conversion of A to B takes place when
energy is supplied & reaction results in
increase in free energy & decrease in
entropy e.g biosynthesis of large
molecules
The reaction is said to be endergonic &
does not occur spontaneously
The reaction consumes energy in
biological systems.
What determines whether this reaction takes
place spontaneously or not?

Principles of thermodynamics help

to solve this question
 Thermodynamics is the study of energy
transformations as applied to all physico-
chemical systems including biological
Bio-energetics is the study of energy
conversion in biological systems
 First law: energy can neither be created nor
destroyed but can be converted from one
form to the other e.g chemical energy into
mechanical energy in muscle contraction
 Second law: all energy transformations result
in an increase in disorder (entropy)
How can these endergonic reactions be made
thermodynamically favourable?

√ Energy coupling
Is the use of an exergonic process to
drive an endergonic process
Is the process of making
thermodynamically unfavourable
reactions to become favourable
The coupling of exergonic reactions
of nutrient breakdown to the
endergonic processes is required to
maintain the living state
 Every metabolic pathway must be a thermodynamically
favoured process.
 HOWEVER, many individual reactions are unfavourable.

 Unfavourable reactions require coupling.

 Reactions are said to be coupled when a

thermodynamically unfavorable (usually endergonic)
reaction (when free energy value is positive) is driven by
coupling the reaction to a thermodynamically favorable
(usually exergonic) reaction (when free energy value is
negative).

 For example, a biosynthetic (endergonic) reaction may be

coupled with the hydrolysis of ATP for it to proceed: A +
ATP + H2O = B + ADP +Pi + H (ΔGº = negative)
 For a pathway to proceed from beginning to end, the
overall G must be negative. Therefore, there is a need to
I. decrease energy need
12
How do living things acquire the energy
needed for these functions?

Phototrophs: acquire free energy from

sunlight
Chemotrophs: oxidize organic
compounds to make ATP
Metabolism
(syn = intermediary metabolism)
 Is the totality of the chemical reactions in the cell
catalyzed by enzymes

 Sequence of enzymatic reactions that produce

specific products are called metabolic pathways
 The intermediates, substrates or products of these
enzyme-catalyzed reactions are called metabolites

 There are 3 types of metabolic pathways:

- Catabolism or catabolic pathway: “down”
- Anabolism or anabolic pathway: “up”
- Amphibolism or amphibolic pathway: “cross road”
Metabolic pathways
Catabolism: is the breakdown of large
molecules to small molecules. It yields energy
Anabolism: is the formation of big molecules
from small molecules. It consumes energy
Amphibolism: is the pathway that involves in
both breakdown and build up of molecules. It
is described as “cross road” between anabolic
and catabolic pathways e.g CAC

NOTE: The 3 metabolic pathways do not occur

in isolation but interact with one
another
Main aims of catabolism: is to form
1. ATP
2. Reducing power e.g NADPH in HMP shunt
3. Building blocks for biosynthesis
 ATP is generated by oxidation of fuel molecules
like glucose, fatty acids & amino acids
 The most common intermediate in most of these
oxidations is acetyl CoA which is completely
oxidized to CO2 by CAC with concomitant
formation of NADH & FADH2
 These electron carriers transfer their high energy
potential electrons to the respiratory chain
 The subsequent flow of electrons to O2 leads to
the pumping of protons across the inner MT
membrane
 Then this proton gradient is used to synthesize
ATP
3 Main pathways for energy production

Glycolysis
Citric
Acid Cycle
Oxidative phosphorylation

Building blocks for biosynthesis

Acetyl CoA : biosynthesis of F.as, PGs
& Choles
 Pyruvate
CAC intermediates
Ecclesiastes 3:1-3
‘To everything there is a season, and a time to every purpose under the heaven.
A time to be born, and a time to die; a time to plant, and a time to pluck up that
which is planted. A time to kill, and a time to heal; a time to breakdown, and a
time to build up’

Anabolic & catabolic pathways are

different & separated
This separation enables both biosynthetic
& degradative pathways to be
thermodynamically favourable at all times
Anabolic pathway which is endergonic is
made exergonic by coupling it to the
hydrolysis of ATP
The separation also helps in the
effectiveness of metabolic control
Metabolic pathways are irreversible
 They have large negative free energy changes to
prevent them running at equilibrium.
 If two pathways are interconvertible (from 1
to 2 or 2 to 1), the two pathways must be
different!
 There is need to control the amounts of either 1 or
2 independent of each other.

A
1
2

Y X
Methods of metabolic pathway
control/regulation
Feed back inhibition (-ve feed
back)
- Products of pathway controls its own
rate of synthesis
- Occurs in the committed (1st) step.
Committed step is the 1st irreversible
reaction in a metabolic pathway
- Its advantage is prevention of
intermediate accumulation
A E1 B E2 C E3 D

23
Methods of metabolic regulation (contd)

 Feed forward activation (+ve feed back)

- Here, metabolites produced early in pathway
activates an enzyme later in the pathway
- Also prevents accumulation of intermediates
A E1 B E2 C E3 D E4 E
 Allosteric interactions (Allosteric
activators and inhibitors)
- Involves activities of certain enzymes of
metabolic pathway rather than the amount
of substrate available (as in 1& 2 above)
- Enzymes that catalyze irreversible reactions
are likely control sites.
24
 Covalent modifications: involves
- Addition of phosphoryl group via protein
kinase
- Removal of phosphoryl group via protein
phosphatase
It advantage is that it allows metabolic
pathways to be rapidly swithed on or off by
very low concentrations of triggering signals.
It lasts longer than allosteric interactions

 Enzyme levels
- Both the amount (quantity) and activity
(activation and inhibition) are controlled
- Rates of synthesis & degradation of many
regulatory enzymes are altered by hormones
25
Compartmentation

- Some enzymes and substrates are

restricted to cytosol and certain
organelles so as to make the
substrates and enzymes available
together in right place

26
 Biological Oxidation,
Respiratory Chain and Oxidative
Phosphorylation
 Biological Oxidation
 All biological processes on this planet are directly or
indirectly affected by oxidation-reduction reactions

 In photosynthetic organisms, solar energy is

converted to chemical energy

 Photosynthetic organisms use this chemical energy to

sustain life during the day and to produce
carbohydrates from CO2 that can be stored as
metabolic fuel for use at night

 All other organisms obtain chemical energy from their

environment, which in many cases, means consuming
the organic materials produced by photosynthetic
organisms and using them as metabolic fuel for
aerobic respiration

 Bioenergetics
- is the term used to describe the processes involved
during energy conversion reactions in living systems
 Livingorganisms need a constant input of energy to
avoid death as long as possible

 The reason for this is that life is dependent on the

maintenance of a highly ordered steady state called
homeostasis which requires energy

 An organism that is at equilibrium with its

environment is no longer alive

 Organisms must maintain a steady state that is far

from equilibrium in order to survive

 For example, the concentration of glucose needed

to sustain life in a cactus is much higher inside the
cactus than it is in the surrounding desert, and it
requires the process of photosynthesis to provide
the energy needed to keep it alive
 Similarly, the concentration of sodium chloride is
lower inside the cells of whale than it is in the
surrounding ocean

 It is the whale's diet of shrimp and plankton that

provide the chemical energy required to maintain a
safe intracellular sodium chloride concentration

 When an organism can no longer maintain

homeostasis using these energy conversion
processes, the intracellular concentration of water,
essential ions, and macromolecules, begin to
equilibrate with the surroundings and the organism
dies

 The reason all living organisms need an input of

energy (which can be stored), is to delay reaching
equilibrium with the environment as long as possible
 Energy conversion in living systems is required for
three types of work that maintain the steady state:
1. chemical work in the form of macromolecular
biosynthesis of organic molecules

2. osmotic work to maintain a concentration of

intracellular salts and organic molecules that is
different than the extracellular matrix and

3. mechanical work in the form of muscle

contraction

 The conversion of hydrogen to helium by

thermonuclear reactions in the sun, and the
subsequent release of energy in the form of visible
light, is called solar energy

 Solar energy provides all of the energy required for

the two types of organisms that inhabit this planet,
photosynthetic autotrophs and heterotrophs
 Photosynthetic autotrophs use solar energy to oxidize
H2O and generate chemical energy that is used to maintain
homeostasis during daylight hours

 Photosynthetic autotrophs also use this chemical energy to

convert atmospheric CO2 into carbohydrate (C6H12O6) which
is a storage form of energy used at night

 The process of oxidizing H2O to capture chemical energy

and generate O2 is called photosynthesis, whereas, the
conversion of CO2 to C6H12O6 is carbon fixation

 Heterotrophs, which includes all non-photosynthetic

organisms, are dependent in one way or another on
photosynthetic autotrophs as a source of chemical energy
(carbohydrate) which is used as metabolic fuel for aerobic
respiration

 Importantly, the production of O2 by photosynthetic

autotrophs through the oxidation of H2O is critical for
aerobic respiration because O2 is the terminal electron
acceptor in this process
 Metabolic Redox Reactions

Oxidation-Reduction = (Redox)

 Both photosynthesis and aerobic respiration interconvert

energy using a series of linked oxidation-reduction
reactions in which electrons are transferred from a
molecule of higher electrochemical potential to one of
lower electrochemical potential

 Oxidation Is the Loss of electrons and Reduction Is the

Gain of electrons (OIL RIG)

 A series of linked oxidation-reduction reactions, called

redox reactions, transfer electrons from one compound
to another

 Since electrons do not exist free in solution, a reduced

compound becomes oxidized when it transfers an
electron to an oxidized compound that becomes reduced

 The importance of redox reactions in biochemical

processes is that chemical work can be performed using
 The chemical energy is obtained by burning (oxidation) of
foodstuffs like carbohydrates, fats and proteins

 When these foodstuffs are degraded, there is production

of reduced co-enzymes like FADH2, FMNH2, NADH + H+ and
NADPH + H+

 For metabolic pathways to proceed or continue there must

be re-oxidation of these reduced co-enzymes since the
continuation of metabolic pathways depends on the
availability of FMN, FAD and NAD+.

 The oxidation of FMNH2, FADH2 and NADH + H+ by

respiratory O2 with simultaneous production of H2O is the
final stage of biological oxidation reactions

 The oxidation of FADH2 and NADH + H+ by O2 accompanies

release of energy which is used for the formation of ATP
and a small amount of energy is released as heat
 NADPH + H+ is re-oxidized back by biosynthetic pathways that
require reduced NADP+

 The transfer of hydrogen atoms (or electrons) of FADH2 and

NADH + H+ to respiratory O2 is a stepwise process and
sequence of specific carriers are needed to carry these
electrons or hydrogen atoms.

 During the transfer of electrons from reduced co-enzymes to

O2, the carrier molecules undergo coupled oxidation-reduction
reactions because whenever one carrier is oxidized the other
carrier is simultaneously reduced

 Therefore the electron transfer in biological system involves

coupled oxidation-reduction reactions

E.g FADH2 + 2Fe3+ → FAD + 2e- + 2H+ …………(Oxidation)

2Fe3+ + 2e- → 2Fe2+ ………………. (Reduction)

 Biological oxidation deals with the use of respiratory O2 in the

body
 Redox reactions involve electrons donors (reductants)
and electrons acceptors (oxidants)

 When a substrate can exist as an oxidant and as a

reductant, it becomes a redox couple or a redox pair

e.g. FAD/FADH2, 1/2O2/H2O, FE3+/FE2+.

Reduction (Redox) potential

 It is a measure of the tendency of a redox couple to
donate electrons in a redox reaction

 When determined under standard conditions (at 25oC and

reactant concentration of 1M), the redox potential
becomes standard redox potential. It is usually
designated as Eo and always measure in volts

 A redox couple with Eo < 0 implies a greater tendency to

donate electrons (i.e oxidation) while the one with Eo > 0
has greater tendency to accept electrons (i.e reduction).
The ∆Eo’ of a Redox reaction

 The change in the redox potential for a redox reaction i.e

∆Eo’ is usually the difference in the standard redox
potentials of the participating couples

∆Eo’ = Eo’ (oxidant) – Eo’ (reductant)

 Redox potentials of some biologically important redox

pairs are given below:

1/2O2/H2O +0.82
Cytochrome c +0.25
FAD/FADH2 -0.18
NAD+/NADH + H+ -0.32
Α-ketoglutarate/isocitrate -0.38
Acetyl-CoA/pyruvate -0.48
For example,
Calculate ∆Eo’ of the following biological redox reaction:
NADH + H+ + 1/2O2 → NAD+ + H2O

∆Eo’= [+0.82-(-0.32)] volt

= 0.82 + 0.32 volt = 1.14volt

Electron transfer and free energy

 When electrons flow from electronegative redox pair
towards electropositive redox pair free energy is liberated

The amount of free energy liberated when electrons move

from one redox pair to another is given by the equation:
∆Go’ = -nf∆Eo’
Where ∆Go’ = standard free energy change in calories
n = number of electrons transferred
f = faraday constant = 23.6Kcal
∆Eo’ = change in redox potential

 Assignment: Calculate standard free energy change for the

above reaction.
 Most of the metabolic energy generated in living cells
during metabolism results from processes that
abstract electrons from a donor molecule, channel
them through an electron transport chain, and finally
deliver them to an acceptor molecule

 These carrier molecules carry or transfer electrons

from reduced coenzymes (which serve as donor
molecules) like NADH or FADH2 to final electron
acceptor O2

 This process is called electron transport chain (ETC) or

respiratory chain, and together with another process
called oxidative phosphorylation are metabolic
processes seen in cellular respiration

 The site of these 2 coupled processes is the inner

membrane of mitochondria
Overview of Oxidation of Glucose
The Relationship of the Electron Transport System
and Oxidative Phosphorylation to other Metabolic
Pathways
 Mitochodria: brief review
- Are ellipsoidal in shape i.e. they look like a jelly bean

- Has length of about 7 microns & diameter of about 1

micron

- Consist of outer & inner membranes, cristae and matrix

- Cristae are invaginations of inner membrane into the

matrix

- It is called power house of the cell because it is

responsible for the production of energy in the form of
ATP
 The position of a particular carrier molecule in the
respiratory chain depends on its redox potential

 The components of respiratory chain are arranged in the

order of increasing redox potential

 Starting components have negative redox potential and

terminal components have positive redox potential

 Therefore, in the respiratory chain, electrons flow from

negative to positive

 Redox potential is defined as e.m.f of a redox pair when

oxidant and reductant are present in 1M concentration

NAD FMN Co Q Cyt b Cyt c1 Cyt c Cyt a Cyt

a3 O2
-0.32V -0.12V +0.04V +0.07V +0.23V +0.25V +0.29V
 In the respiratory chain, electrons flow from NAD to
cytochromes via CoQ and then from cytochromes to
molecular O2

 CoQ also collects electrons from FAD

 The transfer of electrons from substrates to NAD and FAD

is catalyzed by dehydrogenases

 At the electronegative end of respiratory chain, NAD

linked dehydrogenases like malate and glutamate
dehydrogenases catalyze transfer of electrons from
substrate to NAD directly but electrons from substrates
like pyruvate, and α-ketoglutarate are transferred via FAD

 In contrast some FAD linked dehydrogenases like

succinate and glycerol-3-phosphate dehydrogenases
catalyze transfer of electrons from substrate to CoQ of
respiratory chain because their redox potentials are more
positive
 Complexes of Respiratory Chain

 The respiratory chain consists of 4 complexes and 3 mobile

carriers

 The complexes are complex I, II, III and IV while the mobile
carriers are NAD, CoQ and cytochrome c

 All the complexes are integral membrane proteins

 Each complex is involved in oxidation and each complex

accepts electrons from mobile carrier and pass electrons to
another mobile electron carrier

 NOTE THAT: Complexes I, III and IV are proton pumps while

complex II does not pump protons but it serves as a physical
link to citric acid (or Krebs’) cycle

 Complex V is not part of the respiratory chain but the enzyme

complex that carries out the oxidative phosphorylation reaction
i.e. the actual conversion of ADP to ATP
Complete oxidation of glucose by molecular oxygen can be described as:
C6H12O6 + 6O2  6CO2 + 6H2O DGo’=-2823 kJ/mol
This can be broken down into two half-reactions with the transfer of
electrons
i. C6H12O6 + 6H2O  6CO2 + 24H+ +24e-
ii. 6O2 + 24H+ + 24e-  12H2O
- 12e- from the oxidation of glucose are not transferred directly to O 2
- They go to NAD+ and FAD to form 10NADH and 2FADH 2
- These are reoxidized, passing their electrons to the electron-
transport chain to reduce O2 to H2O causing the mitochondrion to
create a proton gradient
- This pH gradient is used to drive the synthesis of ATP via
oxidative phosphorylation
 Complexes of Electron Transport Chain

 Complex I – NADH CoQ reductase or NADH

dehydrogenase

- Catalyzes the first step of the reactions involved in cellular

respiration

- Has 2 cofactors: FMN and Fe-S cluster

- Electrons collected by NAD to form NADH + H + are

transferred to FMN to give reduced FMNH 2. Electrons are
then transferred to Fe-S cluster and finally to CoQ. So NAD
is oxidized and CoQ is reduced to QH 2

- Note that the flow of 2 electrons from NADH to CoQ leads

to the pumping of 4H+ out of mitochondrial matrix. This is
because the Fe-S cluster involved is 4Fe-4S cluster

 NADH + 5H+ + Q → NAD+ QH + 4H+

 Complex II – Succinate CoQ reductase or Succinate
Dehydrogenase

- It contains FAD and Fe-S clusters as cofactors

- It transfers electrons from succinate and glycerol-3-phosphate to

CoQ via FAD and Fe-S cluster

- Succinate dehydrogenase is one of the enzymes in the TCA cycle

- It is the only TCA cycle enzyme embedded in the mitochondrial inner

membrane

- The conversion of succinate to fumarate results in reduction of the

enzyme-bound FAD

- The oxidation of the reduced flavin requires the donation of electrons

to Coenzyme Q

- i.e. the enzyme cannot carry out the reaction if the electron
transport is non-functional

- Succinate dehydrogenase does not pump protons

 Complex III – CoQ-cyt c reductase
- It is a complex of cyt b, Fe-S and cyt c1

- Complex III accepts electrons from Coenzyme Q (and

therefore, indirectly, from both Complex I and Complex
II)

- Complex III contains several heme prosthetic groups

- It transfers electrons from QH2 to cyt c via cyt b, Fe-S and

cyt c1

- As a result of this, CoQ is oxidized and cyt c is reduced

- Like Complex I, Complex III is also a proton pump

 QH2 + 2cyt c ox + 2H+ matrix → Q + 2cyt c red + 4H+ cytosol

 Complex IV – Cytochrome c oxidase

- Cytochrome c oxidase, as the name implies, accepts electrons from

cytochrome c

- Catalyzes the oxidation of cyt c and reduction of O 2 to water which is the

final stage of electron transport chain

- The complex has a very high affinity for molecular O 2

- It contains 2 heme a groups (a and a3) and copper ions

- Cytochrome c oxidase is sometimes referred to as the cytochrome a-a3

complex

- During the reduction of O2, 4 electrons are thought to be transferred

with concomitant formation of water. This is because 4 molecules of
reduced cyt c are thought to be involved in the reduction

- cytochrome c oxidase is the only one that is capable of binding oxygen;

all of the others have both of the axial heme-iron binding sites occupied
by amino acid side-chains

 4cyt c + 8H+ matrix + O2 → 4cyt c + 2H2O + 4H+

red ox cytosol
 Other components (Carrier molecules) of electron
transport chain

1. Flavo-proteins (FAD or FMN)

- They are important hydrogen carriers

- They are usually closely associated with iron-sulfur proteins
which make possible the exchange of electrons between
them
- Their Eo’ are higher than that of NAD+/NADH + H+
- They are integral membrane proteins

2. The iron-sulfur proteins (non-heme iron proteins)

- They are electrons carriers

- They are proteins containing iron-sulfur centers
- They use non-heme iron to accept electrons and transfer
electrons thereby changing from Fe 2+ (ferrous) to Fe3+
(ferric) states
- They are integral membrane proteins
- Iron and sulfur are present in equimolar amounts
3. Cytochromes
- They are components of electron transport chain
present in mitochondria
- They are b, c, c1, a, and a3
- They contain heme-iron which usually undergoes
valence changes from Fe2+ and Fe3+ states
- Only the heme-iron on cytochrome a/a3 is capable of
binding molecular O2, CO and other potential ligands
while other cytochromes can not.
- The cytochromes except cyt c are integral membrane
proteins. Cyt c is a peripheral protein
- They are involved in transfer of electrons from
ubiquinone to cytochrome oxidase
4. Coenzyme Q (Ubiquinone)

- Coenzyme Q is a non-protein electron carrier located in

the inner mitochondrial

- Coenzyme Q can accept electrons from Complex I and II

and donates the electrons to Complex III

- The quinones are a group of yellow-to-red pigments

derived from o- or p-benzoquinone

- The quinones of the inner mitochondrial membrane are

designated ubiquinones (UQ), due to their ubiquitous
occurrence, while the term plastoquinone (PQ) is being
5. Protein-bound compounds

- It is the last component of respiratory chain which

transfers electrons to molecular O2. During the process,
copper changes valence between Cu+ and Cu2+
 Oxidative Phosphorylation

- Glycolysis and the citric acid cycle yield NADH and FADH 2

- Both these electron carriers are energy-rich molecules

because their electrons have a high transfer [redox]
potentials

 Oxidative phosphorylation is the process of converting this

high redox potential into energy-rich ATP molecules.

- This process, together with the reactions that form the

electron carriers is often called respiration

- During the transfer of electrons in the respiratory chain,

energy is released because electrons flow from
electronegative NAD to electropositive O2

- The energy so released is used for the formation of ATP

from ADP and phosphate (Pi)

- The process accounts for over 80-90% of ATP produced in

the body
 Mechanism

- The concept of oxidative phosphorylation looks simple but the

mechanism is a little bit complex

- The flow of electrons from NADH or FADH2 to O2 through protein

complexes in the mitochondrial inner membrane leads to the
pumping of protons out of the mitochondrial matrix

- This results to imbalance of protons concentration of the

mitochondrial matrix compared to the concentration outside the
matrix, and this generates a gradient known as proton gradient and
a trans-membrane electrical potential is as well generated that
leads to the creation of proton motive force (p.m.f)

- This p.m.f tries to drive proton back into the mitochondrial matrix
through some enzyme/protein complexes

- This flow of proton back into the matrix is what is needed to drive
ATP Synthase to generate ADP and Pi to form ATP

- The mechanism just described is called Chemiosmotic coupling

model or hypothesis and is the most accepted

- There are other mechanisms of oxidative phosphorylation which are

not accepted due to lack of experimental support
 ATP Synthesis

- Formation of ATP from ADP and phosphate using energy

released when electrons flow in the respiratory chain is
catalyzed by membrane bound protein known as ATP
Synthase or F0F1 ATPase

- It is often referred to as another complex V of the

respiratory chain. It is found on the cristae of inner
mitochondrial membrane

- Protons return to the mitochondrial matrix through an

integral membrane protein (of the mitochondrial inner
membrane) known as ATP synthase

- ATP synthase is a multiple subunit complex that binds ADP

and inorganic phosphate and converts them to ATP

- ATP Synthase consists of 2 subunits: F0 and F1 subunits. The

spherical part or head of knob is F1 subunit and is made up
of 5 polypeptide chains while the base of the knob is F 0
- When protons move from outside to inside of inner
mitochondrial membrane F1 subunit catalyzes the
formation of ATP from ADP and Pi using free energy
released

- The stalk of the knob is where antibiotic oligomycin binds

to inhibit oxidative phosphorylation
 Sites of oxidative phosphorylation or ATP synthesis
in the respiratory chain

- ATP synthesis takes place in the inner mitochondrial

membrane and it is associated with 3 complexes of the
electron transport chain

- These complexes are complex I, complex III and complex

IV
 Inhibitors of respiratory chain and oxidative phosphorylation

(a) Compounds which act at complex I (NADH-CoQ reductase) or

at site I
1. Barbiturates which is used as sedative inhibits NADH-CoQ reductase.
So, electron flow is blocked and ATP synthesis does not occur
2. Rotenone (a fish poison) blocks flow of electrons from Fe-S clusters. It
is also used as insecticide
3. Pericidin (an antibiotic)

(b) Compounds which act site II of ATP synthesis (Co-Q cyt c

reductase)
1. Antimycin A (an antibiotic)
2. BAL (British anti lewisite). It is used as therapeutic agent in the cases
of arsenic poisoning

(c) Compounds which act site III (Cytochrome oxidase)

1. Cynanide. It is a powerful poison that inhibits cytochrome oxidase by
combining with cytochrome a3.
2. Carbon monoxide. It is a pollutant present in automobile exhaust
3. Hydrogen sulfide. Its toxicity occurs during oil drilling operation. It is a
part of natural gas
4. Azide e.g sodium azide
 Other inhibitors of oxidative phosphorylation

1. Carboxin. It blocks the transfer of electrons from succinate

to CoQ i.e it inhibits complex II
2. Atractyloside. It blocks movement of ATP and ADP across
inner mitochondrial membrane
3. Oligomycin. It interacts with stalk of knob of ATP synthase
and completely blocks oxidation and phosphorylation
4. Rutamycin. It blocks phosphorylation without uncoupling

 Uncouplers
- These compounds dissociate or uncouple oxidation in
respiratory chain from phosphorylation. So, oxidation takes
place without ATP synthesis. Examples are:

i. 2, 4-dinitrophenol
ii. Dinitrocresol
iii. Pentachlorophenol
iv. Salicylanilides
v. CCCP (Carbonyl cyanide chloromethoxy phenyl hydrazone)
vi. Arsenite. It acts by competing with phosphate for ATP
synthesis