Electron Transport Chain

and
Oxidative Phosporylation

CJW

Learning Objectives & Outcomes

This lecture will emphasize on the following topics;
which the students are expected to understand at the
end of this lecture.

1. Know the course of electron transport chain in

mitochondria
2. Know how the electrical and pH gradient lead to
the production of ATP in oxidative phosphorylation
3. Know the various inhibitors and uncouplers for
them

Review of glycolysis and the TCA cycle

There are three main pathways that generates energy
in the body
Energy production pathways:1. Glycolysis
2. TCA cycle
3. Electron transport chain & oxidative
phosphorylation
All three of them are inter-connected

So far, we have learn two of them

Next is the coupled reaction of electron transport
chain and oxidative phosphorylation

Recap from Glycolysis and TCA cycle

Synthesized in Glycolysis/TCA cycle
NADH, Nicotinamide adenine
dinucleotide
FADH2, Flavin adenine dinucleotide
Energy rich reduced co-enzymes
Utilized in ETC and OP
H+ gradient
ATP synthase
NADH and FADH2 ; get oxidized
ADP get phosphorylated

Review of glycolysis and the

TCA cycle
Electron carrying nucleotides (NADH
and FADH2) generated from
glycolysis and TCA cycle are funneled
to the electron transport chain to
generate MORE ATP

Food generates power

Various sources of food are used to generate ATP
The electrons are being carried as NADH and FADH2
In the end oxygen is reduced

Tales of the Two Systems that Generate Energy

1. Electron transport chain (ETC)
Electron pass through an electron transport chain
Loose free energy, but generates a H+ gradient

2. Oxidative phosphorylation (OP)

H+ pass through ATP synthase; produce ATP

Both processes are highly integrated / coupled

process
All cells that have mitochondria

Mitochondrial structures
Location, location, location!
Outer membrane

Inter-membrane space (H+ accumulates)

Inner membrane
(Electron transport assembly +
ATPsynthase)
Cristae
Matrix

 The chain has 4

multiprotein
complexes (IIV)
Note where NADH /
FADH2 donates the
electron

2
1

Too complex?
4
FMN Flavin mononucleotide
FeS Iron-sulphur protein
Q Coenzyme Q10
Cyt Cytochrome (heme)
FAD Flavin adenine dinucleotide

Oxidation and Reduction

The key to understand this chapter
Reduced
1. Addition of H
2. Removal of O
3. Additional of electrons
Oxidized
1. Removal of H
2. Additional of O
3. Removal of electrons

Reduced Form
(eg. NADH, FADH2)
Oxidized Form
(eg. NAD+, FAD+)

Simpler ?

1
3
2

Chemi-osmotic theory

There is a need for reducing agents

To generate the proton gradient in the
intermembrane region, the mitochondria require a
constant supply of reducing agents (NADH and
FADH2) from Kreb Cycle
And next door (cytosol), they have plenty of NADH
(from glycolysis) transport in via:1. Malate-aspartate shuttle
2. Glycerophosphate shuttle

A deal is a deal: Malate-Aspartate shuttle

Transfer of NADH from cytosol to Mitochon.
Malate is used to shuttle H+ into Mitochon. (like TCA)

E:

Ketoglutarate
transporter

E:

E:

E:
Glutamateaspartate
transporter

A deal gone bad: Glycerolphosphate shuttle

Short-changed, as NADH has been replaced by FADH2
FADH2 has less reducing power compared to NADH
NADH = 3 ATP ; FADH2 = 2 ATP

Everyone is an accountant
6 ATP
4 ATP
6 ATP

2 ATP

18 ATP

4 ATP

2 ATP

Famous trick question (Ans: Campbell & Reece)

NADH = 3 ATP
FADH2 = 2 ATP

Keeping up with the demand

As ATP synthase generates
more ATP, it has to be
exported out
And theres a demand for
ADP

E: adenine nucleotide
transporter / translocase (2)
Inorganic phosphate is
required as well; phosphate
transporter (1)

Uncouplers & inhibitors

Uncouplers
Allow protons to flow back into the matrix of the
mitochondria from the intermembrane region,
without ATP synthesis, and with the consumption of
oxygen
Inhibitors
Inhibitors blocks the transfer of electron within the
electron transport chain (ETC), denying the pump of
proton at each complexes, thus reducing the proton
gradient between the intermembrane region and the
matrix of the mitochondria

Uncouplers (saboteur)
ETC and OP is a coupled reaction
The most evil one. What do they do to the ETC?
It allows proton to flow back into the matrix of the
mitochon., without making ATPs
Rendering the whole electron transport and
reduction of oxygen, POINTLESS
Eg:
1. Dinitrophenol

2. Long-chain fatty acids

3. Valinomycin is a K+ ionophore disrupt the
electrochemical gradient, also prevent ATP synthesis

Dinitrophenol
Amphipathic
Increases the permeability of H+ through the innermembrane
Reducing the electrochemical potential (proton
gradient)
Short-circuiting the ATP synthase
Leaked proton can be oxidized ( H2O) without
phosphorylating ADP

Are you slim enough ?

Dinitrophenol uncouples oxidative phosphorylation,

causes release of calcium from mitochondrial stores
and prevents calcium re-uptake. This leads to free
intracellular calcium and causes muscle contraction
and hyperthermia.

Inhibitors
Each type of inhibitor block a specific site of the electron transfer
3

Complex IV

Complex III

Complex I

RTC e- transfer inhibitors

Complex I
Rotenone pesticide/insecticide
Amytal (Amobarbital) barbiturate class, sedative,
analgesic
Complex III
Antimycin produced by bacteria (Streptomyces), as
fish poison
Complex IV
Cyanide used primarily in industries
CO used primarily in industries
released during fires

RTC non-e- transfer inhibitors

ADP-ATP phosphorylation (E: ATP synthase)
Blocks phosphorylation
Oligomycin antibiotic; result in high levels of lactate
(due to regeneration of NAD+, in glycolysis)
pyruvate lactate
ADP-ATP transport (E: translocase)
Blocks ADP-ATP transportation cytosol-matrix
A-trac-ty-lo-cide - from Mediterranean thistle, a
glycoside
Bong-kre-kate - antibiotic produced by Pseudomonas
cocovenenans

Inhibitors & uncouplers

e- carrier (reduced) before [INHIIBITOR] after e- carrier (oxidized)

Cyanide poisoning

Jewelery & textile industry

usually attempted suicide
Symptoms:1. neurological impairment
2. severe acidosis
Antidote:1. sodium thiosulphate
2. hydroxocobalamin

Cyanide poisoning
Cyanide poisoning is also seen in:1. Survivors from fires in buildings with fittings that
produces HCN gases upon combustion
2. Sewerage workers
3. Petroleum refineries
4. Gold extraction

Cyanide poisoning
LD=1.5mg /kg
If 70kg = 105 mg = 0.1 g
Sad but true. Used for gas chambers mass killing;
industrial or household fires
Best murder weapon: HCN, colorless, faint bitter almond
scent (but most ppl cant detect it by smell)
[read more LMS]
Firstline: Amyl nitrite

Antidote biochemistry
Amyl nitrate stimulates the formation of methemoglobin
which has high affinity for cyanide forming
cyanomethemoglobin

Sodium thiosulphate induces the E: Sulfur transferase

activity; which converts the formed
cyanomethemoglobin to thiocyanate (excreted in urine)
Hydroxocobalamin (has a cobalt ion) which has high
affinity for cyanide; binding of cyanide, forms
cyanocobalamin (Vit B12)

Cyanide can be extracted from the pits of apricot as amygdalin; 1 kernal = 0.5 mg cyanide

References
Harper RK, Granner DK, Mayes, PA, & Rodwell, VW
(2003). Harpers Illustrated Biochemistry (26th Ed).
McGraw-Hill. Chapter 12: The respiratory chain and
electron transport chain.
Champe, PC., Harvey, RA & Ferrier, DR. (2008).
Lippincotts Illustrated Review, 4th Ed. LWW. Chapter 30
RNA structure, synthesis and processing.
Lieberman & Marks (2009). Marks Basic Medical
Biochemistry: A clinical approach, 3rd Ed. Chapter 14
Transcription: Synthesis of RNA