FACULTY OF MEDICINE

BMS 1153 Cell Metabolism

Biochemistry LECTURE # 1
Energy Metabolism
 LEARNING OUTCOMES
At the end of this session the student would be able
to:

Describe the process of ATP formation along with

H2O from NADH , FADH2 and molecular O2
Discuss the relationship between movement of
electrons through the complexes and pumping of
protons generating electrochemical gradient and role
of ATP synthase enzyme in generating ATP from ADP
List the inhibitors and uncouplers of the oxidative
phosphorylation process

May 15, 2017 2

 KEY WORDS
BIOENERGETICS
OXIDATIVE PHOSPHORYLATION
ELECTRON TRANSPORT CHAIN
HIGH ENERGY PHOSPHATES
ENERGY TRANSFORMATION ( energy changing from
one form to another)
ENERGY TRANSDUCTION (is the conveyance of energy
from one electron (a donor) to another (a recipient)
REDUCING POTENTIALS (One way to quantify whether a
substance is a strong oxidizing agent or a strong reducing
agent ).
CHEMIOSMOTIC MODELThe production of ATP from ADP is
coupled to the transfer of electrons in the electron transport
chain.
PROTON GRADIENT ( difference in concentration of the
protons across the inner membrane of mitochondria).
PROTON MOTIVE FORCE (chemical potential energy due
to the proton gradient).
TRANSMEMBRANE REGION
Life is dependent on energy transformations.
Living organisms survive because of
exchange of energy within and without.
Energy is required for mechanical work or for
other anabolic processes. Bonds are broken
and High energy bonds (~P ) are made.
The production of High energy bonds allows
transduction of usable energy.
Chemical bonds are broken and made as
part of the exchange and transformation of
energy in living organisms.
Bioenergetics

The part of biochemistry

concerned with the energy
involved in making and breaking
of chemical bonds in the
molecules found in biological
organisms.

It can also be defined as the

The energy in glucose is used to produce ATP via
the reactions of glycolysis, cellular respiration,
and the electron transport system .

BETA OXIDATION

ATP
92% of ATP is produced by Ox-Phos in
mitochondria.
Compounds with high energy bonds are said to have high
group transfer potential.

For example, Pi may be spontaneously cleaved from ATP for

transfer to another compound (e.g., to a hydroxyl group on
glucose).
NH2
ATP
adenosine triphosphate N
N

G = -7.3kcal/mole
O O O N N

-O P O P O P O CH2
adenine
O
O- O- O- H H
H H
phosphoanhydride OH OH
bonds (~) ribose
Phosphocreatine (creatine phosphate)
another compound with a "high energy" phosphate
linkage, is used in nerve & muscle for storage of ~P
bonds.

Creatine Kinase catalyzes:

Phosphocreatine + ADP ATP +
Creatine
This is a reversible reaction, though the
equilibrium constant slightly favors
phosphocreatine formation.
G = -10.3kcal/mole
Phosphocreatine is produced when ATP levels are high.
O
When ATP is depleted CH3in muscle, phosphate is
during exercise O
transferred from H

O P phosphocreatine
N C Nto ADP,
CHto replenish
2 C ATP.
O
O NH2+

phosphocreatine
Oxidative phosphorylation
Transduction of energy from the oxidation
of NADH and FADH2 via the ETC system to
the phosphorylation of high energy bonds
of ATP.
The process by which the energy from
fuel oxidation ( NADH and FADH2 from
Glycolysis, beta-oxidation and TCA ) is
transduced to high energy phosphate
bonds of ATP via the electron transport
chain system in the inner mitochondrial
membrane.
 Electron transport chain
The assembly of protein complexes in the
inner membrane of mitochondria that
participate in the energy transduction process
via cascade of oxidation-reduction steps.
NADH
ADP + Pi ATP + H2O
-320mV
SYNTHASE

DGo
1.14volts
PROTONMOTIVE FOR
-52.4kcal

+ 820mV e- + O2 + 2H+ H2O

Oxidative phosphorylation occurs in
three
1. phasesof energy-rich molecules
Production
NADH and FADH2.
2. Electrons from NADH and FADH2 are
then passed along the electron
transport chain to the terminal
electron acceptor O2. The free energy
released in electron transport is
captured by coupling it to the export of
protons across the mitochondrial inner
membrane.
3. Finally, the free energy of the
electrochemical proton gradient is
used to activate ATP synthase,
 Oxidative phosphorylation
Three phases:
(a)Production of NADH and FADH2
(b)Electron transport - the transfer
of electrons from NADH and FADH2
to O2 through a series of
intermediate electron carriers, with
concomitant export of protons from
mitochondria and
(c)ATP synthesis - utilization of the
electrochemical potential energy of
the proton gradient ( proton motive
force) to synthesize ATP from ADP +
 Electron Transport Chain

The transfer of electrons from NADH

to O2 releases substantial free
energy.
This energy is harvested by coupling
electron transport to the export of
protons out of the mitochondrial
matrix, against an electrochemical
gradient.
To capture the energy most
efficiently, electrons are passed
Components of Electron
Transport Chain

ATP synthase
III IV
I

II
 THE ELECTRON
TRANSPORT CHAIN
The electron carriers of the
electron transport chain are
organized into four multi protein
complexes.
These complexes are connected by
two mobile electron carriers,
ubiquinone and the Fe - heme
containing protein cytochrome C.
Ubiquinone diffuses freely in the
hydrocarbon core of the inner
mitochondrial membrane.
Cytochrome C (Cyt C) diffuses
 Electron Transport Chain
(ETC)
Assembly of large protein complexes ( I, II,
III and IV ), Cyto C and a hydrophobic
coenzyme Q (ubiquinone with 10
isoprenoid units).
Complex I is e-acceptor for NADH.
Complex II is e-acceptor for FADH2.
These components are arranged
sequentially embedded in the inner
mitochondrial membrane.
ATP synthase is a multi subunits complex
with Fo as proton channel and F1 is the
enzyme ATPase.
18
COMPLEX 1 COMPLEX III COMPLEX IV ATPase complex

Fo
Co Q CYTO C

F1
intermembrane

Fo

F1

Mitochondrial matrix
 The F0F1 ATP synthase

F1 domain faces towards the inside of the

mitochondrial matrix, and carries out ATP
synthesis.
F0 domain resides in the inner
mitochondrial membrane, and is a proton
channel.
Appears as a dense array of studs on the
inner mitochondrial membrane when
examined under the electron microscope.
Coupled only by the proton motive force
generated by ETC.
 OXIDATIVE PHOSPHORYLATION
Electron Transport Chain complexes in
mitochondrial inner membrane. A series of
Oxidation- reduction reactions along the
component of the etc.
e- e- e-
e-
NADH complex I CoQ complex III
Cyto C
e- e-
complex IV O2

The energy from the ETC is utilized to

translocate H+ from matrix to inter-
membrane region. The gradient created
induces influx of H+ , which activate ATP
synthase and thus :
ADP + Pi ATP + H2O
 Complex I, NADH
NADH dehydrogenase, 880 kD, 34
subunits.
Complex II,
FADH2 succinate
CoQ
dehydrogenase
(TCA cycle), 140
kD, 4 subunits.
Complex III, cytochrome
reductase, 250 kD, 10
subunits.

Cyto C

Complex IV, cytochrome

oxidase, 160 kD, 10 subunits.

O2
 Oxidation / Reduction

Oxidation = the loss of e-

and results in the loss of
hydrogen atoms together
with one or two e- ( also
known as dehydrogenation
reactions). Or gain of oxygen
atom.

Reduction = gain in e- and

results in gain in hydrogen
 THE CHEMIOSMOTIC
MODEL
Explains the ATP synthesis mechanism of the
electron transport chain in the mitochondrial
membrane.
Explains the flow of e- (from NADH or FADH2 )
along the components of the electron transport
chain translocate protons from matrix to the inter-
membrane region, creating proton gradient across
the inner membrane.
The pH gradient activates ATPase complex and open
proton channel. The influx of proton produce energy
to synthesize high - energy phosphate bond of ATP.
ADP + Pi ATP + H2O
 PROTON MOTIVE FORCE
The energy from the e- cascade is not
directly utilized for ATP synthesis . The e-
energy is used for the translocation of H+
from matrix to trans-membrane region of
mitochondria, thus creating proton
gradient across the inner membrane of
mitochondria.
The proton motive force generated
activates ATPase.
SUMMARY OF OXPHOS
The energy from fuels oxidation is carried in the form
of NAD(H) and FAD(H2).
The ETC oxidizes NAD(H) and FAD(H 2) and transfers e-
sequentially to O2 , which is then reduced to H 2O.
The e- energy induces proton transport across the
mito membrane, creating a proton gradient.
The proton motive force activates ATP synthase
(FoF1 ATPase) :
ADP + Pi -> ATP + H2O.
Net yield of OxPhos is only 3 OR 2.5 mole of ATP
per mole of NADH and 2 OR 1.5 mole of ATP per
mole of FAD(H2) oxidized (the rest energy for
transport and heat).
G = -52.3kcal/mole of NADH .
SUMMARY of Ox-Phos.
Reduced cofactors (NADH/FAD2) donate e- via the
components of ETC to O2 to form water.
90% O2 inhaled is used in this aerobic respiration.
Energy from e- is harnessed to translocate proton
from matrix to the transmembrane region.
Electrochemical gradient generated across the inner
mito membrane, change the pH and change inner
membrane potential.
The proton motive force drives back proton into
matrix via proton channel in Fo/F1 ATPase.
Change in conformation activates ATPase and
catalyzes formation of ATP.
Respiration is coupled to ATP synthesis.
Inhibitors , Uncouplers & Diseases of Oxphos

Inhibitors of oxphos: Cyanide, Rotenone,

Amytal, Carbon monoxide, Oligomycin,
Valinomycin, Antimycin C, Atractyloside
insecticide rotenone

Uncoupler - Dinitrophenol

Oxphos diseases are degenerative diseases ,

caused by gene mutations in either
mitochondrial or nuclear DNA that encodes
proteins or enzymes required for normal
 There are six distinct types of poisons which
may affect mitochondrial function:
1) Respiratory chain inhibitors (e.g. cyanide, antimycin,
rotenone)
2) Phosphorylation inhibitors (e.g. oligomycin)
3) Uncoupling agents (e.g. dinitrophenol)
4) Transport inhibitors (e.g. atractyloside, bongkrekic
acid) either prevent the export of ATP, or the import of raw
materials across the the mitochondrial inner membrane.
5) Ionophores (e.g. valinomycin,) make the inner
membrane permeable to compounds which are ordinarily
unable to cross.
6) Krebs cycle inhibitors (e.g. arsenite, aminooxyacetate)
 Action of natural Uncoupler
(UCP-1) protein in
thermogenesis of Brown
Adipose tissues (BAT)
UNCOUPLER

Proton leak back into matrix No ATP synthesis

Mitochondria of brown adipose tissue with its

characteristic protein, uncoupling protein-1 (UCP-1)
- (Anti-obesity potential ?).
 THERMOGENESIS IN BROWN ADIPOSE TI
Only 30% energy from Ox-Phos is used for ATP synthesis ,
others for transport and heat.
Uncoupling proteins - mitochondrial inner membrane
proteins that dissipate the proton gradient before being used
to provide the energy for oxidative phosphorylation.
Natural Uncoupler, UCP-1 allows respiration , no ATP
synthesis, energy is dissipated as heat.
UCP1, also known as thermogenin play a role in cold exposure
or hibernation, because the energy is used to generate heat
(thermogenesis) instead of producing ATP.
Uncoupling proteins are enhanced by thyroid hormone,
norepinephrine, epinephrine, and leptin.
Human UCP3 is mainly expressed in skeletal muscle, which
plays an important role in energy homeostasis and substrate
oxidation.
Therefore, UCP3 is a good candidate gene for treating obesity.
 Chemical uncoupler 2,4-dinitrophenol
(DNP). DNP acts as a protonophore, allowing
protons to leak across the inner
mitochondrial membrane and thus bypass
ATP synthase.

Salicylic acid is also an uncoupling agent and

will decrease production of ATP and increase
body temperature if taken in excess.
 References

Lippincotts Illustrated Biochemistry.

Text Book of Biochemistry for Medical Students by
DM Vasudevan.
Harpers Biochemistry 26th edition.