The Respiratory Chain

(ETC)
OP

Tewodros Shibabaw-20
 Objectives
1. label the anatomy of mitochondria
2. Discuss respiratory chain and OP in
relation to the chemiosmotic model.
3. Identify the components of the
respiratory chain and ATP
synthase complex, and describe their
roles.
4. Define respiratory control, inhibiting
and uncoupling, and
 Mitochondrial Anatomy
 Elliptical “egg
shaped “
 Two membranes
 Matrix
 The Location Of Enzymes Responsible For
Electron Transport And Oxidative
Phosphorylation.

In inner
membrane
Impermeable
to ions and
most other
compounds
 Mitochondrial Transport
Uniport – Moves a Single substance such as

Ca++,H+.

Symport- Moves Two substance in the same

direction like

Pyr: H+; PO4+3:H+

Antiport –Moves Two substance in opposite

direction such as
 (An Overview)
Biological oxidations are catalyzed by
intracellular enzymes. The purpose of oxidation is to
obtain energy.

Electron Transport: Electrons carried by reduced

coenzymes (NADH or FADH2) are passed sequentially
through a chain of proteins and coenzymes (so called
ETC )to O2 .

Oxidative Phosphorylation: Coupling e-

Transport (Oxidation) and ATP synthesis
(Phosphorylation) .
 The Electron Transport
Chain…
 Again Each complex can accept electrons
and then transfer them to other complexes
through mobile carriers, Ubiquinone (Q) and
WhatC.are
Cytochrome the source of
carriers ?
What are the electron
carriers ?
ter-membrane space
I II III IV ATPase
4H 4H 2H
+ 0H + +
+

H+
NADH O2 H2 O
NAD+succinate
fumarate ADP

ATP
Mitochondrial matrix
 Electron Carriers
The transfer of electrons is not directly to
oxygen but through Coenzymes
NAD+

FMN There are 2 sites of entry for

FeS
electrons into the ETC:

FAD FeS ubiquinone NAD+ or FAD : Both are coenzymes for

Cyt b
dehydrogenase enzymes

ubiquinone

FeS Cyt c1 Cyt c Cyt a Cyt a3

1/2 O2
Ubiquinone
Other names and abbreviations:
Coenzyme Q, CoQ, Q NAD+
O
CH3O CH3 FMN

CH3 FeS
CH3O (CH2 CH C CH2)nH
O coenzyme Q FAD FeS ubiquinone

2 e + 2 H+ Cyt b

OH
ubiquinone
CH3O CH3

FeS
CH3
Most often n = 10
CH3O (CH2 CH C CH2)nH
OH Q + 2 e- + 2 H+  QH2.
coenzyme QH2
Coenzyme Q (CoQ, Q or ubiquinone) is lipid-

soluble. It dissolves in the hydrocarbon core of a

membrane.

The only electron carrier not bound to a protein.

It can accept/donate 1 or 2 e-. Q can mediate e-

transfer
Cytochromes
NAD+

FMN Proteins that accept electrons

from QH2 or FeS
FeS

Ultimately transfers the electrons

FAD FeS ubiquinone
to oxygen
Cyt b

ubiquinone

FeS Cyt c1 Cyt c Cyt a Cyt a3

1/2 O2
Cytochromes
Cytochromes are electron carriers containing
hemes . Hemes in the 3 classes of cytochrome (a,
b, c) differ in substituents on the Porphyrin ring.

Some cytochromes(b,c1,a,a3) are part of

large integral membrane protein complexes.

Cytochrome c is a small, water-soluble

protein.
 Composition of Respiratory Chain Complexes

No. of Prosthetic
Comple Name Protei Groups
ns
xComplex I NADH 46 FMN,
Dehydrogen 9 Fe-S
ase cntrs.
Complex Succinate- 5 FAD, cyt
II CoQ b560,
Reductase 11 cyt3bFe-S
, cyt bL,
Complex CoQ-cyt c H
centrs.
III Reductase cyt c1, Fe-
Complex Cytochrom 13 cytSa, cyt a3,
Rieske
IV CuA, CuB
e Oxidase

You have to know what complex mean ?

 Mitochondrial complexes Components
of the ETC ,
And NAD
ATP
+ synthase.

FMN
I NADH Dehydrogenase
FeS

FAD FeS ubiquinone

II
Cyt b
Succinate
Dehydrogenase
ubiquinone Cytochrome Oxidase

FeS Cyt c1 Cyt c Cyt a Cyt a3

III IV
1/2 O2
CoQ-cyt c Reductase
upport for this order of events
Energetically favorable.

Spectra: the absorption spectrum

Specific inhibitors.

Assay of individual complexes.

Order and Reduction Potentials E0
NAD+
-0.32

FMN
-0.3
FeS

FAD FeS ubiquinone +0.0

45
+0. Cyt b
03 +0.07
ubiquinone 7
+0. 29 +0. 55
FeS Cyt c1 Cyt c Cyt a Cyt a3

+0. 22+0. 25
1/2 O2

+0.8
 Drugs/Chemicals that inhibit the ETC
NAD+
Rotenone helps natives of
Amytal FMN the
Rotenone I Amazon rain forest catch
FeS
fish!
FAD FeS ubiquinone

II CN- CO
Cyt b
Binding tightly to the
ubiquinone
ferric
Antimycin Form (fe3+) of a3
Cyt c1 Cyt c Cyt a Cyt a3
A FeS

III IV
1/2 O2
 Mitchell's Chemicosmotic Theory
1. Protons are translocated from the matrix into inter
membrane space as e-s move from reduced
coenzymes to O2 couple reaction

2. The consequence of the aforementioned vectorial

movement is generation of electrochemical gradient
Or Release free energy and conserving the free energy
of fuel oxidation as a transmembrane electrochemical
potential
3. E.C.G is converted to energy content of ATP by
proton
Back flow via Fo to phosphorylation of ADP ( ATP
synthase). couple reaction
he electron transport chain

4H 4H 2H
+ + +
Redox potential E0 - Measures whether a pair is likely to
donate or accept electrons
(affinity for electrons relative to H+/H2
= 0.000 volt )
21
omplex-I= NADH to Ubiquinone.
Complex I, also called NADH:Ubiquinone

Oxidoreductase or

NADH dehydrogenase, is a large enzyme.

High-resolution electron microscopy shows

Complex I to be L-shaped,
 2. Complex II: Succinate to Q
SDH , the only membrane-bound enzyme in the citric

acid cycle .

Although smaller and simpler than Complex I, it

contains five prosthetic groups of two types and

four different protein subunits .

 3. Complex III: Q to

Cytochrome c
Complex III, also called cytochrome bc1

complex or ubiquinone: cytochrome c

Oxidoreductase, couples the transfer of electrons

from QH2 to Cyto. C with the pumping of protons .

4.Complex IV: Cytochrome C to
O2
In the final step of the respiratory chain, Complex IV,
also called cytochrome oxidase, carries electrons
from Cyt. C to molecular oxygen, reducing it to
H2O.
 5.ATP Synthase
ATP synthase (F0F1ATPase), the enzyme that
generates ATP, is a
multi-subunit enzyme containing an (F0) and
headpiece (F1) that
project into the matrix .
The headpiece is composed of three subunit pairs.
Each subunit
contains a catalytic site for ATP synthesis.
 The protons have a thermodynamic tendency to
return to the matrix =Proton-motive Force

The proton move back into the matrix through

the FoF1ATP synthase driving ATP
synthesis. Coupled !!!!!!!!
Small Molecule Shuttles
ATP-ADP Translocase
Phosphate Translocase
uses 1 H+.
ATP Currency Exchange Ratios of NADH and FADH 2
Experimentally 3 H+ are required to 1 ATP when they flow back
down the electrochemical proton gradient through the ATP
synthase complex, and 1 H+ is needed to transport 1Pi
molecule into the matrix.
Cont…
We can now see where the ATP currency exchange
ratios

of ~2.5 ATP/NADH and ~1.5 ATP/FADH2 come

from:

Oxidation of NADH starting at complex I

yields:
10 H+/4 H+ = 2.5 ATP
Oxidation of FADH2 starting at complex II
yields:
6 H+/4 H+ = 1.5 ATP for FADH2
 Summary of Reactions
Complex I:
NADH + 5H+N + Q → NAD+ + QH2 + 4H+P
Complex II:
FADH2 + Q ==> FAD + QH2
Complex III:
QH2 + 2 cyt c1 (ox) + 2 H+N → Q + 2 cyt c1 (red) + 4 H+P
Complex IV:
2 cyt c (red) + 4H+N + ½ O2 → 2 cyt c (ox) + 2 H+P + H2O
 Steps in Respiration
Electron Donors
=Organic Compounds (Glucose preferred)
Electron Carriers Plus Mobile
Carriers
=NAD to NADH Q to QH2
=FAD to FADH Cyt.C
Electron Acceptors-Terminal
=O2 to H2O
Phosphorylation Reactions
=ADP to ATP
Chemiosmotic Reactions
 Heat generation
The uncoupling protein providing an
alternative route for protons to reenter the
mitochondrial matrix, causes the energy conserved
by proton pumping to be dissipated as heat.

2,4-dinitrophenol/ Aspirin
 ATP synthase inhibitor -
Inhibitors of The
Oligomycin
Respiratory Chain
Uncoupling agents

Natural Uncoupler

=Thermogenin

Chemical Agents =2,4-

dinitrophenol
 Uncouplers Inhibit The Respiratory
Chain
Thermogenin is a Natural Uncoupler
found in the mitochondria of brown fat in
Hibernating animals and infants.

Thermogenin is a membrane protein that permits the

organism to keep warm through metabolism without
having to utilize ATP for movement.

Under such conditions, up to 90% of ATP

derived from fatty acid oxidation in these tissues
Cold stimulates the release of Norepinephrine
from the sympathetic nerve endings. As a result,
a lipase is activated that releases fatty
acids for oxidation. Which types of lipase
iOpener Quiz
1 Which of the following ETC complexes contains a
non-heme copper

A) Site
2 The target I respiratory
in the B) Site II chain
C) Site
for III
cyanide
D) Site is
IV
A) Site I B) Site II C) Site III
4 In OP ,H2O is formed. Where do the O2 atoms in
D) Site IV
the H2O come from?
A) Lactate B)
molecular oxygen
C) carbon dioxide D) glucose