Bioenergetics

MAHPARA GONDAL
PHARM D
MS PHARMACEUTICAL CHEMISTRY

RASHID LATIF COLLEGE OF PHARMACY

BIOENERGETICS
 Bioenergetics is a field in biochemistry that concerns [energy]
flow through living systems. This is an active area of biological
 research that includes the study of the transformation of energy in
living organisms and the study of thousands of different cellular
 processes such as cellular respiration and the many other 
metabolic processes that can lead to production and utilization of
energy in forms such as ATP molecules.
PRINCIPLE
 In a living organism, chemical bonds are broken and made as part of the
exchange and transformation of energy. Energy is available for work (such as
mechanical work) or for other processes (such as chemical synthesis and 
anabolic processes in growth), when weak bonds are broken and stronger bonds
are made. The production of stronger bonds allows release of usable energy.
 So principle of bioenergetics based on laws of thermodynamics:
 First law of thermodynamics and enthalpy
 Second law of thermodynamics and entropy

 In photosynthesis, autotrophs can produce ATP using light energy. Heterotrophs

 must consume organic compounds. These are mostly carbohydrates, fats, and 
proteins. The amount of energy actually obtained by the organism is lower than
the amount present in the food; there are losses in digestion, metabolism, and 
thermogenesis.[1]
CONT…
 Living organisms produce ATP from energy sources via 
oxidative phosphorylation. The terminal phosphate bonds of ATP
are relatively weak compared with the stronger bonds formed
when ATP is broken down to adenosine monophosphate and
phosphate and then dissolved in water. Here it is the energy of
hydration that results in energy release. An organism's stockpile of
ATP is used as a battery to store energy in cells, for intermediate
metabolism. Utilization of chemical energy from such molecular
bond rearrangement powers biological processes in every
biological organism.
OXIDATIVE PHOSPHORYLATION 
 (or OXPHOS in short) is the metabolic pathway in which cells use enzymes to
oxidize nutrients, thereby releasing energy which is used to reform ATP. In
most eukaryotes, this takes place inside mitochondria. Almost all 
aerobic organisms carry out oxidative phosphorylation.

 During oxidative phosphorylation, electrons are transferred from 

electron donors to electron acceptors such as oxygen, in redox reactions. These
redox reactions release energy, which is used to form ATP. In eukaryotes, these
redox reactions are carried out by a series of protein complexes within the inner
membrane of the cell's mitochondria. These linked sets of proteins are called 
electron transport chains.
CONT…
 The energy released by electrons flowing through this electron transport chain is used
to transport protons across the inner mitochondrial membrane , in a process called 
electron transport. This generates potential energy in the form of a pH gradient and an 
electrical potential across this membrane. This store of energy is tapped by allowing
protons to flow back across the membrane and down this gradient, through a large
enzyme called ATP synthase; this process is known as chemiosmosis. This enzyme
uses this energy to generate ATP from adenosine diphosphate (ADP), in a 
phosphorylation reaction. This reaction is driven by the proton flow, which forces the 
rotation of a part of the enzyme; the ATP synthase is a rotary mechanical motor.
 Although oxidative phosphorylation is a vital part of metabolism, it produces 
reactive oxygen species  such as superoxide and hydrogen peroxide, which lead to
propagation of free radicals, damaging cells and contributing to disease and, possibly, 
aging (senescence). The enzymes carrying out this metabolic pathway are also the
target of many drugs and poisons that inhibit their activities.
MITOCHONDRIAL ELECTRON TRANSPORT CHAIN

 Electron transport chain (ETC) is a series of highly organised

oxidation-reduction enzymes whose reactions can be represented as:
 Reduced A + Oxidised B ⇔ Oxidised A + Reduced B.

 The ETC is localised in the mitochondria. The outer membrane of

mitochondria is permeable to most of the small molecules. There is
an intermediate space which presents no barrier to passage of
intermediates.
 The inner membrane shows a highly selective permeability. It
has transport systems only for specific substances such as ATP,
ADP, pyruvate, succinate, α-ketoglutarate, malate and citrate etc
CONT….
 The enzymes of the electron transport chain are
embedded in the inner membrane in association
with the enzymes of oxidative phosphorylation.
 The most accepted sequence of electron carriers
in the mitochondria is as follows:
CONT…
 The chain actually consists of a series of redox couples,
at each step electrons flow from the reductant of a redox
couple with more negative redox potential to the oxidant
of the next redox couple having a more positive redox
potential.
DETAIL STRUCTURE AND FUNCTIONS
OF ELECTRON TRANSPORT CHAIN (ETC)

 The electron transport chain in the mitochondrial

membrane has been separated in 4 (four)
complexes or components as follows:
 • Complex I: NADH-CoQ reductase
 • Complex II: Succinate-CoQ reductase
 • Complex III: CoQ-cytochrome C reductase
 • Complex IV: Cytochrome C oxidase.
COMPLEX I: NADH-COQ REDUCTASE

 This system has two functions:

 • Electron transfer
 • Acts as a proton pump.
 The system catalyses transfer of two electrons from NADH
to small lipid soluble CoQ via FMN and Fes clusters.

 From FMN.H2 electrons are transferred to a group of Fes

proteins. Fe atoms of FeS protein oscillate between Fe++
and Fe+++. The electrons are then transferred to CoQ.
CONT….

 The process is accompanied by pumping of protons

from mitochondrial matrix into intermembrane space.
 • Permits one ATP Formation (Site I).
 Note: Upto CoQ, H is transferred. But from CoQ
onward only e– is transferred, 2H+ goes into the
medium.
COMPLEX II: SUCCINATE-COQ REDUCTASE

 Flow of electrons from succinate to CoQ occurs via FAD.H2.

 Succinate + CoQ ___________> Fumarate + CoQ.H2

 Standard reduction potential for transfer of electrons from

FAD.H2 to CoQ is + 0.113V (much lower than +0.420 V
energy change for the reaction of complex I). The small
energy change does not allow “succinate-CoQ reductase”
system to pump protons across the mitochondrial membrane,
hence this protein complex does not contribute to proton
gradient. Hence no ATP is formed.
CONT…
 Note: CoQ is the electron acceptor in the reaction
catalysed by NADH-CoQ reductase (Complex I)
and succinate-CoQ reductase (Complex II). The
electrons received are subsequently transferred
from CoQ.H2, a lipid soluble mobile electron
carrier to CoQ-Cyt.C reductase (Complex III).
COMPLEX III: COQ-CYT.C REDUCTASE

 Functions as
 • Proton pump, and
 • Catalyses transfer of electrons
 This system catalyses transfer of electrons from
CoQ.H2 to Cyt-c via Cyt-b and Cyt-c1.
 The electrons from CoQ.H2 is first accepted by
Cyt-b566 and then transferred to Cyt-b562,
which reduces Co.Q.H2 CoQ
CONT….
 Fe+++ accepts electron and is oxidised to Fe++.
 The system also acts as a proton pump. It is
believed that 4 (four) protons are pumped across the
mitochondrial membrane during the oxidation.

 The energy change permits ATP formation (Site

II).
COMPLEX IV: CYT-C OXIDASE

 The system functions:

 • As proton pump
 • Catalyses transfer of electrons to molecular O2 to form
H2O.
 This is the terminal component of ETC. It catalyses the
transfer of electrons from Cyt-c to molecular O2 via Cyt-
a,

 Cu++ ions and Cyt-a3. The flow of electrons is as follows:

CONT…
 Role of Cu Ions: Cu atom adjacent to cyt. haem a is Cu-A
(Sub unit II) and Cyt-haem a3 is close to CuB (Sub unit I).
From Cyt-c, the electrons are transferred to haem-a-CuA
cluster and then haem a3-CuB cluster.
 The system also acts as a proton pump, it pumps two protons
into intermembrane space. The energy change permits ATP
formation (Site III) between cyt a3 and molecular O2.
 Note: Cyt c does not form a part of any complexes. It is
mobile and acts as a shuttle between complex-III and
complex-IV to transfer e– (electron).
CONT…
 At three sites free energy released per electron
pair transferred is sufficient to support the
phosphorylation of ADP to ATP which requires
about 8 KCal/mole.
OXIDATIVE PHOSPHORYLATION
 The process of oxidative phosphorylation is closely associated with the
functioning of the electron transport chain.
 In oxidative phosphorylation ATP is produced by combining ADP and
Pi with the energy generated by the flow of electrons from NADH to
molecular oxygen in the electron transport chain. There are three sites
in the respiratory chain where ATP is formed by oxidative
phosphorylation.
 These sites have been proved by the free energy changes of the various
redox couples. Since hydrolysis of ATP to ADP + Pi releases around 7.3
K. Cal/mole, the formation ATP from ADP + Pi requires a minimum
of around 8 KCal/mole. The formation of ATP is therefore not possible
at the sites where free energy released is less than 8 KCal/mole.
SITES OF ATP FORMATION:
 There are three sites in the respiratory chain
where ATP can be formed.
• Site I: This involves the transfer of electrons from
NADH –CoQ. Obviously this step is omitted by
succinic dehydrogenase whose FADH2 prosthetic
group transfer its electrons directly to CoQ
bypassing NADH. This step is blocked by
piericidin, rotenone, amobarbital, certain drugs
like chlorpromazine, guanethidine.
CONT…
 Site II: This involves the transfer of electrons from
Cyt-b– Cyt-c1. This step as well as the previous one is
bypassed in oxidation of L-ascorbate whose electrons
are directly transferred to Cyt-c. This step is blocked by
Antimycin A, Hypoglycaemic drug like phenformin.

 Site III: Transfer of electrons from Cyt-a3 to molecular

oxygen which is blocked by CO, CN, H2S, and azide.
MECHANISM OF OXIDATIVE PHOSPHORYLATION

 Three major proposals for the mechanism of

oxidative phosphorylation have been considered.
The synthesis of ATP is carried out by a
molecular assembly in the inner mitochondrial
membrane. This enzyme complex is called
Mitochondrial ATPase or H+-ATPase. It is also
called ATPsynthase.
THEORIES

 The three hypothesis do make use of the

information available on ATP synthase.
 A. The chemical coupling hypothesis: This is
developed from the concept of a high energy
intermediate common to both electron transport
and phosphorylation of ADP. However, such
intermediate has not been identified so far.
CON…
 The conformational coupling hypothesis:
According to this hypothesis the mitochondrial
cristae undergo conformational changes and these
changes in architecture of the mitochondrial
cristae reflect the changes in the different
components of the electron chain to one another.
It is believed that these conformational change
represents the formation of high energy state
CONT…
 Chemiosmotic theory: This is the most accepted
view of oxidative phosphorylation postulated by
Peter Mitchell in 1961. Mitchell’s chemiosmotic
theory postulates that the energy from oxidation of
components in the respiration chain is coupled to the
translocation of hydrogen ions (Protons, H+) from
the inside to the outside of the inner mitochondrial
membrane. Each of the respiratory chain
complexes I, III and IV acts as a proton pump
CONT…
 The inner membrane is impermeable to ions in
general but particularly to protons, which accumulate
outside the membrane, creating an electrochemical
potential difference across the membrane (ΔμH+).
This consists of a chemical potential (difference in
pH) and an electrical potential. The electrochemical
potential resulting from the asymmetric distribution
of the hydrogen ion is used to drive the mechanism
responsible for the formation of ATP