Respiration

Phase 2 of respiration:
(ATP generation from oxidative
phosphorylation)
ILOS:

 Define electron transport chain

 Demonstrate components of electron transport chain
 Enumerate sources of NADH+H+ and FADH2 to ETC
 Describe shuttle pathways
 Discuss chemiosmotic hypothesis
 Enumerate factors that control the ATP synthesis
 Describe oxidative phosphorylation inhibitors
 Discuss thermogenesis in brown adipose tissue
 Mention levels of ATP production
 Mention mitochondrial DNA mutation and disease
Mitochondrial oxidation—the electron
transport chain
 def :
 The electron transport chain (ETC) converts the
electrons stored as reduced intermediates NADH and
FADH2 into a proton motive force (PMF) across the
inner mitochondrial membrane (IMM).
 site :
The ETC (except for cytochrome c) is located in the inner
mitochondrial membrane.
> ETC converts the electrons stored as reduced intermediates NADH
and FADH2 into a proton motive force (PMF) across the inner
mitochondrial membrane (IMM).
Complexes of electron transport
chain :
The electron transport chain are organized in 4 complexes;
Complex I. Complex II, Complex III, and Complex IV.
Structure of complexes : enzyme and hydrogen or
electron carrier .
Complex I : It contains NADH dehydrogenase enzyme, FMN as
hydrogen carriers, and iron- sulfur center.
It take 2 hydrogen atoms ( 2 e and 2 H+) from NADH+H+, which
is then pass to FMN, and lastly the two electrons pass to
iron-sulfur center and then to coenzyme Q .
 Complex II : It contains FAD, succinate dehydrogenase and iron –
sulfur center.Succinate dehydrogenase is the only enzyme of Kreb,s
cycle that is attached to the inner mitochondrial membrane. Electrons
move from succinate to FAD, then to iron-sulfur center and finaly to
coenzyme Q to produce coenzyme QH2..
Coenzyme Q : It is a lipophilic and mobile electron 
carriers .
It receives electrons from both complex I and 
complex II. So, link these complexes with cytochrome
c1 of complex III.
 Complex III : It contains cytochrome b and
cytochrome c1. The iron of heme is in ferric state.
Iron acts as electron acceptor and donor by
changing its valency. Electrons of cytochrome c1 are
transferred to cytochrome c.
 Cytochrome C : is a mobile electron carrier that
diffuses through the inner mitochondrial membrane
shuttling electrons from c1 of complex 111 to CuA
of complex 1V.
 Complex IV : It contains cytochrome a and
cytochrome a 3 .It contains copper proteins. Electrons
are passed from cytochrome c to cytochrome a, a3
and finally to O2. Complex IV reacts directly with
oxygen , so called cytochrome oxidase. At complex IV ,
the electrons, O2 and protons are brought together
and H2o is formed.
•Three of the four complexes are also proton pumps: for
each pair of electrons, complex I extrudes 4H+ from the
matrix; complex III, 4H+; and complex IV, 2H+. Complex II
does not pump protons when it transfers electrons from
FADH2 to coenzyme Q

• Thus, for each NADH oxidized, 10H+ are extruded; and,

for each FADH2, 6H+

• The ultimate electron acceptor is molecular oxygen, which

is reduced to water.
Sources of NADH+H+ and FADH2 to
ETC :
The main sources are :
 β–oxidation of fatty acid
 Krebs cycle
Both occur in mitochondrial matrix adjacent to ETC
in the inner mitochondrial membrane.
 NADH+H+ is also formed cytoplasmically e.g. in
glycolysis.
But There is no direct pathway for NADH to cross the
inner mitochondrial membrane (IMM) to enter the ETC.
So How NADH can effectively cross the membrane ??
Malate/ Aspartate shuttle:
When cytoplasmic NADH is low, the glycerol-3-phosphate
shuttle may be used. Electrons enter the ETC at the level
of FADH2 and so get less ATP per original NADH than
with the malate/aspartate shuttle.
ATP synthesis: The Chemiosmotic Theory

 Chemiosmotic hypothesis
explains how the free
energy generated by the
transport of electrons by
ETC is used to produce
energy.
 Complex V : It is the site
of biosynthesis of ATP. It
contains two subunits, F0
which span the inner
mitochondrial membrane
and F1 which protrude into
the mitochondrial matrix.
ATP synthase enzyme is a
part of this complex.
proton pumps
 Transfer of electrons from one complex to another complex (oxidation )
is accompanied by liberation of energy which is used to pump proton
from mitochondrial matrix to the inter membrane space. Complex I ,
Complex III , and Complex IV acts as proton pump.

 Three of the four complexes are proton pumps: for each pair of electrons,
complex I extrudes 4H+ from the matrix; complex III, 4H+; and complex IV,
2H+. Complex II does not pump protons when it transfers electrons from
FADH2 to coenzyme Q

 Thus, for each NADH oxidized, 10H+ are extruded; and, for each FADH2,
6H+
 Each ATP formed uses four protons. Hence 1 NADH = 2.5 ~ 3
ATP, 1 FADH2 = 1.5~ 2 ATP

 The ultimate electron acceptor is molecular oxygen, which is reduced to

water.
 Oxidative-phosphorylation coupling :
The energy released from oxidation is used to
pump protons which accumulate in the inter
membrane space creating electrochemical gradient
outside the mitochondria. The electrochemical
gradient drive the protons to renter the matrix
by passing through a proton channel in the F0
driving rotation of F0 and conformational change
in F1 and binding of ADP to P to form
ATP(phosphorylation ).
Respiratory control
 Electrons cannot flow through the ETC unless
ADP is simultaneously phosphorylated to ATP.

 The most significant controlling factor for

electron flow is the availability of ADP for
conversion to ATP.
 Oxidative Phosphorylation Inhibitors:

Electron Directly inhibit electron transport, so block ATP Rotenone: complex I inhibitor.
synthesis. Malonate:Competitive inhibitors of
transport
succinate dehydrogenase (complex II).
inhibitors Antimycin A: complex III
inhibitor.
Cyanide, carbon monoxide, azide
inhibit complex IV.
ATP synthase Directly inhibit mitochondrial ATP synthase, by Oligomycin
blocking the flow of protons through the
inhibitors Fo subunit causing an increase of•
proton gradient.
No ATP is
produced because electron transport stops.

Uncoupling I•ncrease permeability of membrane, causing a 2,4-Dinitrophenol (used illicitly for

decrease of• proton weight loss),
agents gradient.ATP synthesis Aspirin (overdose),
stops, but electron transport continues. Thyroxine in large doses.
It allows oxidation to occurs but prevents Thermogenin in brown fat (has more
phosphorylation (ATP formation ). The energy is mitochondria than white fat).
produced in the form of heat giving the
sensation of hotness .
Thermogenin (uncoupling protein UCP ):
 It is physiological uncoupler found in brown adipose
tissue that function to generate heat.
 There are different types of UCP:
UCP1 ,UCP2,UCP3,UCP4,and UCP5 .
Green tea polyphenols and UCP : Green tea
polyphenols have anti-obesity effects by increasing the
expression of UCP2 which enhance thermogenesis.
Levels for ATP production : .

 1-Substrate levels phosphorylation : The energy

which is used for biosynthesis of ATP is derived
from substrates containing high energy bonds. There
are 3 examples for substrate level phosphorylation:
 2-Oxidative phosphorylation : The enegy used for
formation of ATP is derived from oxidation .This
occurs in electron transport chain.
 Mitochondrial DNA
Definition :Circular DN present in
mitochondria.
Function : Expression of enzymes of
electron transport chain.
 So, any mutation in mitochondrial DNA will
affect respiratory chain and defect in energy
production.
Mutations in mitochondrial DNA

1- Mitochondrial myopathy :
 Causes: mutation in mtDNA of skeletal muscle
 Effects : deficiency of one of ETC enzymes >> decrease
ATP
 C/p:
Ms weakness
Ms cramps
Severe fatigue after minimal exercise
 Investigations :
Ms biopsy
Molecular analysis
 2-LHON
Lebers Hereditry Optic Neuropathy
c/p: degeneration of optic nerve leading to
sudden blindness
 3-MELAS:
Mitochondrial Encephalopathy Lactic Acidosis And
Stroke
Due to deficiency of NADH-Q reductase enzyme

N.B: Mitochondrial inheritance is considered as a maternal

inheritance.
References:
 Wilkins R,Cross S, Megson L and Meredith D
(2011):Oxford Handbook of Medical Sciences Second Edition

 Tao Le, Vikas Bhushan Matthew Sochat, Yash Chavda,

Kimberly Kallianos, Jordan Abrams, Mehboob Kalani
and Vaishnavi Vaidyanathan (2019): FIRST AID for the
USMLE Step 1.

 Sandra K. Leeper-Woodford and Linda R. Adkison,

(2016): Lippincott Illustrated Reviews: Integrated Systems.