University of Central Punjab

A “W4” Category University

Assignment no. 01
Subject: Biochemistry
Topic: Oxidative phosphorylation
Submitted By: Rida Zainab
MIF18BSCH0113
Session 2018-2022
Submitted To: Prof Afzia Anwar
For fulfill the requirement for semester 6th

University of Central Punjab, Multan Campus

Faculty of sciences

Oxidative phosphorylation:

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The process of synthesizing ATP from ADP and Pi coupled with the electron transport chain
is known as oxidative phosphorylation.

Energy is released when electrons are transported from higher energy NADH/FADH2 to
lower energy O2. This energy is used to phosphorylate ADP. This coupling of ATP synthesis
to NADH/FADH2 oxidation is called oxidative phosphorylation. Oxidative phosphorylation
is responsible for 90 % of total ATP synthesis in the cell.

Introduction to oxidative phosphorylation

Oxidative phosphorylation is the major source of ATP in aerobic organisms. It is linked with
the mitochondrial electron transport chain. Oxidative phosphorylation takes place during
respiratory chain. Three ATP-molecules are formed during three steps of the respiratory
chain. This process can express by following equation:

NADH + + 3ADP + 3Pi + 1/202 —> NAD+ + 4H20 + 3ATP

The Pi is an inorganic phosphate. The molecular mechanism of the oxidative phosphorylation

is associated with the respiratory chain. These respiratory chains are present in the inner
membrane of mitochondria. The mechanism of oxidative phosphorylation is chemiosmosis. It
is a process that uses membranes during redox reaction for ATP production.

General steps of oxidative phosphorylation

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Step 1:  Generating a Proton Motive Force

 The hydrogen carriers (NADH and FADH2) are oxidized and release high energy
electrons and protons
 The electrons are transferred to the electron transport chain, which consists of several
transmembrane carrier proteins
 As electrons pass through the chain, they lose energy – which is used by the chain to
pump protons (H+ ions) from the matrix
 The accumulation of H+ ions within the intermembrane space creates an
electrochemical gradient (or a proton motive force)

Step Two:  ATP Synthesis via Chemiosmosis

 The proton motive force will cause H + ions to move down their electrochemical
gradient and diffuse back into matrix
 This diffusion of protons is called chemiosmosis and is facilitated by the
transmembrane enzyme ATP synthase
 As the H+ ions move through ATP synthase they trigger the molecular rotation of the
enzyme, synthesizing ATP

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Step Three:  Reduction of Oxygen

 For the electron transport chain to continue functioning, the de-energized electrons
must be removed
 Oxygen acts as the final electron acceptor, removing the de-energized electrons to
prevent the chain from becoming blocked
 Oxygen also binds with free protons in the matrix to form water – removing matrix
protons maintains the hydrogen gradient
 In the absence of oxygen, hydrogen carriers cannot transfer energized electrons to the
chain and ATP production is paused.
The chemiosmotic theory

Explaination

The chemiosmotic theory was developed by the British biochemist, Peter Mitchell, to
explain the mechanism of oxidative phosphorylation in mitochondria (and
photophosphorylation in chloroplasts). He was awarded the Nobel Prize in 1978. The
processes of oxidative phosphorylation is as follows.

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The NADH produced in the Krebs cycle passes electrons to the respiratory chain and
becomes re-oxidized to NAD+. The electrons are transported down the chain to oxygen
releasing energy, the reduction of oxygen forms water. Thus, oxygen is the terminal electron
acceptor of the mitochondrial respiratory chain.

Respiratory chain

The oxidation/reduction reactions of a respiratory chain is

The red rectangle is the respiratory chain. The arrows show oxidation/reduction (electron transport)
reactions. A, B, C, D are oxidation/reduction components (in reality, there are more).

The respiratory chain can also be represented by a series of conventional equations:

Ared + Box  Aox + Bred

Bred + Cox  Box + Cred

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 Cred + Dox  Cox + Dred

Oxidation/reduction components

The oxidation/reduction components, A, B, C etc include haem groups (containing iron

atoms), flavins, ubiquinone, copper atoms and clusters of iron and sulphur atoms, each can
be reduced by accepting electrons and oxidized by donating them. The pathway of electron
transport down the respiratory chain is complicated but well understood. The
oxidation/reduction components are associated with large protein complexes which traverse
the inner membrane, and cytochrome c, which is a smaller protein located at the outside
surface of the inner membrane. The structures of the respiratory chain proteins and the
positions of their oxidation/reduction components are inferred using X-ray crystallography.

Pathway or procedure of oxidative phosphorylation

Electron transport through the complexes of the respiratory chain causes uptake of H+ from
the matrix at the inside surface of the inner membrane, and the release of H+ at the outside
surface. Protons are effectively pumped outwards across the inner membrane using energy
released in the electron transport reactions.

The movement of H+ makes the outer aqueous region positive and slightly acidic relative to
the matrix of the mitochondrion. This difference between the two sides of the membrane is
called the protonmotive force, and it can drive protons back across the membrane through
the ATP synthase enzyme. This protein acts as a generator, turned by the force of the
hydrogen ions diffusing through it, down their electrochemical gradient. Proton flow
through this enzyme harnesses the potential energy stored in the hydrogen ion gradient and
causes a rotation of protein subunits in the ATP synthase which leads to ATP synthesis
from ADP and phosphate.

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The generation of the protonmotive force and ATP synthesis occur even when the outer
mitochondrial membrane is removed thus the outer membrane does not have a role in
oxidative phosphorylation.

ATP produced in the mitochondrial matrix by oxidative phosphorylation is exported into the
cell cytoplasm by a transporter protein located in the inner membrane. The same protein
continuously returns ADP to the matrix for re-phosphorylation. The total pool of ATP +
ADP in each cell is small and may turn over hundreds of times a day.