LECTURE TOPIC 16: CARBOHYDRATE METABOLISM continued…

 Electron Transport chain

 Uncouplers of the ETC by e.g., thermogenin (uncoupling protein in brown adipose tissue) and the
chemical 2,4-dinitrophenol (DNP) and aspirin overdose
 Oxidative Phosphorylation
 Regulation of oxidative phosphorylation through ATP needs of the cell
Lecture outcomes: By the end of this lecture you should be able to answer the following questions:
1. Components of the ETC: their location and nature as integral & non integral membrane proteins.
2. Understanding of the Redox principle in the electron transport chain.
3. Uncouplers of the ETC: uncoupling of the electron transport chain from oxidative
phosphorylation by e.g., thermogenin (uncoupling protein in brown adipose tissue) and the
chemical 2,4-dinitrophenol (DNP)
4. Oxidative Phosphorylation: definition as opposed to substrate level phosphorylation.
5. Regulation of oxidative phosphorylation through ATP needs of the cell.

For this lecture, Lecture 16, students have to recall Lecture 12: BIOENERGETICS

Structure of a mitochondrion (recall):

Page 1 of 14
Page 2 of 14
Electrons can enter the electron transport chain through either complex I or II. These electrons come from NADH, which is
formed in three reactions of the citric acid cycle. Electrons from FADH2, formed in step 6 of the citric acid cycle, enter the
electron transport chain through complex II. Succinate dehydrogenase, the enzyme in the citric acid cycle that catalyzes the
formation of FADH2 from FAD is part of complex II. The electrons from FADH2 are then transferred to an Fe·S protein. Complexes
III and IV include several iron-containing proteins known as cytochromes. The iron in these enzymes is located in substructures
known as iron porphyrins. Complex III contains cytochromes b and c, as well as Fe·S proteins, with cytochrome c acting as the
electron shuttle between complex III and IV. Complex IV contains cytochromes a and a3 in an enzyme known as cytochrome
oxidase. This enzyme has the ability to transfer electrons to molecular oxygen, the last electron acceptor in the chain of electron
transport reactions. In this final step, water (H2O) is formed.

Page 3 of 14
Page 4 of 14
Page 5 of 14
SUmmarSu

Page 6 of 14
SuuTHE CHEMIOSMOTIC THEORY OF ATP SYNTHESIS (i.e., of oxidative phosphorylation of ADP to form ATP):

Page 7 of 14
The Chemiosmotic theory of ATP synthesis proposed by P. Mitchell is about the coupling of electron
transfer and proton pumping with ATP synthesis. According to this theory, the proton (H+) gradient
(created by the pump complexes I, III, and IV) across the inner mitochondrial membrane is a store of free
energy since the flow of protons (through the Fo channels) back from the inter-membrane space into the
matrix drives ATP synthesis on Complex V also called ATP synthase complex or F1 unit where the
reaction ADP + Pi → ATP occurs. --The potential energy stored in the proton gradient has 2 components:
1
a difference in chemical concentration of protons. Proton concentration is another term for pH, so in other
words, there is a pH gradient across the inner mitochondrial membrane; a difference of 1.4 pH units; 2a
difference in electrical potential. All protons are positively charged (+1) so the excess protons in the inter-
membrane space cause a difference in electrical potential (of about 140 mV) across the inner mitochondrial
membrane. There is therefore a tendency for protons to diffuse back into the matrix (through the F o
channels which span the inner mitochondrial membrane) down the electrochemical gradient.

Page 8 of 14
The F1 subunit couples the flow of protons (from the inter-membrane space into the mitochondrial
matrix) down their concentration gradient to the phosphorylation of ADP. The proton flux through Fo
channels of the enzyme alter the nucleotide-binding properties of the active site, promoting ATP
synthesis. *The Fo channels are integral membrane proteins just as Complexes I, III & IV.

Oxidative phosphorylation is coupled to the electron transport chain and the generation of ATP from
ADP is linked to the oxidation of the coenzymes NADH, FADH2 to NAD+ and FAD respectively in
redox reactions and occurs via the generation of a proton gradient across the inner mitochondrial
membrane. ATP synthesis occurs through oxidative phosphorylation under aerobic conditions: This
oxidative phosphorylation is coupled to the electron transport chain and the generation of ATP is linked
to the oxidation of NADH, FADH2 to NAD+ and FAD respectively in redox reactions.

Oxidative phosphorylation is the 2nd method by which cells synthesize ATP (the 1st one being substrate level
phosphorylation in glycolysis & Krebs cycle). Definition: Oxidative phosphorylation is defined as ATP synthesis
on the ATP synthase complex (Complex 5) on the inner mitochondrial membrane and is coupled to the
oxidation of NADH, FADH2 to NAD+ and FAD respectively in the electron transport chain electron transport chain
and the generation of a proton (H+) gradient across the inner mitochondrial membrane under aerobic
conditions. (Students to include diagrams of the Chemiosmotic hypothesis).

Oxidative Phosphorylation

Each intermediate compound in the electron transport chain is reduced by the addition of one or two

electrons in one reaction and then subsequently restored to its original form by delivering the electron(s) to

the next compound along the chain. The successive electron transfers result in energy production. But how is

this energy used for the synthesis of ATP? The process that links ATP synthesis to the operation of the

electron transport chain is referred to as oxidative phosphorylation.

Electron transport is tightly coupled to oxidative phosphorylation. The coenzymes NADH and FADH2 are

oxidized by the respiratory chain only if ADP is simultaneously phosphorylated to ATP. The currently

accepted model explaining how these two processes are linked is known as the chemiosmotic hypothesis,

which was proposed by Peter Mitchell, resulting in Mitchell being awarded the 1978 Nobel Prize in

Chemistry.

Page 9 of 14
Page 10 of 14
Some of the redox reactions of the electron transport chain:

REGULATION OF OXIDATIVE PHOSPHORYLATION:

Page 11 of 14
 UNCOUPLERS

*Uncouplers are substances that break the connection between the electron transport chain and
oxidative phosphorylation.

Page 12 of 14
Page 13 of 14
*Electron transport is coupled to ATP synthesis on Complex V via oxidative phosphorylation.

-END OF LECTURE 16-

Page 14 of 14