THE RESPIRATORY CHAIN AND

OXIDATIVE PHOSPORYLATION

Cicia Firakania
Medical Faculty of Unswagati Cirebon
2016
By the end of this lecture, the
student should be able to:
1. Understand the chemiosmotic theory
2. Understand the flow of electron in respiratory
chain
3. Understand the mechanism of ATP formation in
electron transport chain
4. Understand the concept of shuttles acrross the
mitochondrial membrane
5. Understand the example of respiratory chain
inhibitors
Definition of oxidation and
reduction reaction
- Oxidation and reduction in terms of hydrogen
transfer

Definitions
Oxidation is loss of hydrogen.
Reduction is gain of hydrogen.
Oxidation and reduction in terms of
electron transfer
Definitions
Oxidation is loss of electrons.
Reduction is gain of electrons.
Introduction
Aerobic organisms are able to capture a far greater
proportion of the available free energy of respiratory
substrates than anaerobic organisms.

Most of this takes place inside mitochondria, which

have been termed the powerhouses of the cell.

Respiration is coupled to the generation of the high-

energy intermediate, ATP, by oxidative
phosphorylation.

A number of drugs (eg, amobarbital) and poisons (eg,

cyanide, carbon monoxide) inhibit oxidative
phosphorylation, usually with fatal consequences.
 Several inherited defects of mitochondria
involving components of the respiratory
chain and oxidative phosphorylation have
been reported.

Patients present with myopathy and

encephalopathy and often have lactic
acidosis.
Cellular Respiration

An ATP-generating process in which an inorganic

compound (such as molecular oxygen) serves as the
ultimate electron acceptor. The electron donor can be
either an organic compound or an inorganic one.
Overview of Cellular Respiration
Mitochondria
Mitochondria have an
outer membrane
that is permeable to
most metabolites
and an inner
membrane that is
selectively
permeable, enclosing
a matrix within.
Specific enzymes in the
mitochondrial compartment
The outer membrane:
acyl-CoA synthetase and glycerolphosphate
acyltransferase
Intermembrane space:
adenylyl kinase and creatine kinase
Inner membrane:
the phospolipid cardiolipin, enzymes of the
respiratory chain, ATP synthase and various
membrane transporters
Role of the respiratory chain of mitochondria
in the conversion of food energy to ATP
Glycerophospate shuttle for transfer of reducing
equivalents from cytosol into the mitochondrion
Malate shuttle for transfer of reducing equivalents
from the cytosol into the mitochondrion
Enzim-enzim dlm siklus Krebs, oksidasi beta dan rantai
respirasi mengumpulkan dan mentranspor ekivalen
pereduksi menuju reaksi terakhir dengan oksigen
membentuk air

Bersama dengan mesin-mesin fosforilasi oksidatif,

menjalankan proses yg menghasilkan energi bebas
yang diperangkap dalam bentuk senyawa fosfat
berenergi tinggi (high-energy phosphate).
Chemiosmotic Theory
How is oxidation coupled to produce of ATP????
Peter Mitchell Propoposed chemiosmotic hypotheses
Chemiosmostic Model
Chemiosmostic Model

The theory postulates that complexes I, III, and IV

act as proton pumps creating a proton gradient
across the membrane, which is negative on the
matrix side. The proton motive force generated
drives the synthesis of ATP as the protons flow back
into the matrix through the ATP synthase enzyme.
Electron Transport System
KOMPONEN DALAM RANTAI RESPIRASI

Ada 4 kompleks protein yang besar yang tertanam

dalam inner membrane mitokondria, tetapi Q
dan Cyt C bersifat mobile.

Elektron mengalir melalui rantai respirasi melalui

reaksi-reaksi redoks dg span 1,1 Volt :
mulai dari NAD+/ NADH O2/ 2 H2O
melewati 3 kompleks besar protein: I III IV
Electron Transport System

Terdiri dari 5 Kompleks

Kompleks I NADH-CoEnzymeQ Oxireductase
Kompleks II - Succinate-Q Oxidoreductase
Kompleks III - Q-Cytochrome c Oxidoreductase
Kompleks IV - Cytochrome C Oxidase
Kompleks V - ATP synthase
Kompleks I IV berfungsi untuk menciptakan
gradien proton yang besar
Kompleks V berfungsi untuk mensintesis ATP
dengan bantuan gradien proton
Complex I NADH-CoEnzymeQ
Oxireductase

elektron
ditransfer/
dipindahkan dari
NADH ke koenzim
Q (Q= ubiquinone).
 Complex II - Succinate-Q
Oxidoreductase

Sebagian substrat dgn

potensial redoks yang lebih
tinggi daripada NAD+/ NADH
(contoh: suksinat) cenderung
melewatkan elektron ke
koenzim Q melalui kompleks II
ini daripada kompleks I.
 Complex III - Q-cytochrome c
Oxidoreductase

kompleks ini
memindahkan
elektron ke
sitokrom c.
Complex IV - Cytochrome c Oxidase

melengkapkan rantai
respirasi dengan
melewatkan elektron
ke O2 , dan direduksi
menjadi H2O
Complex V - ATP synthase
Ko-enzim Q terdifusi dalam membran, sedangkan cyt c adalah suatu
protein terlarut.

Aliran elektron melalui kompleks IIIIIV menghasilkan proton

yg terpompa dari matrik menuju celah antar membran
mitokondria (lihat gambar di slide 9).

Kompleks kompleks Komponen Rantai Pernafasan merupakan

suatu Flavoprotein atau Iron-Sulfur Protein

Flavoprotein adl komponen penting dalam kompleks I dan II.

FMN/FAD
Dpt direduksi pd proses pemindahan 2 elektron membentuk
FMNH2 atau FADH2
Dpt menerima 1 elektron , membentuk semikuinon.
Iron-sulfur protein :
Disebut juga : Nonheme iron proteins, Fe-S
Dijumpai pada kompleks I, II dan III
Dapat mengandung 1, 2 atau 4 atom Fe yg terikat pada atom sulfur
anorganik dan atau melalui gugus SH sistein terikat pd protein.
Fe-S berperan memindahkan elektron, dimana Fe mengalami oksidasi-
reduksi antara Fe2+ Fe3+

Quinon menerima elektron melalui Kompleks I dan II.

Kompleks I :
Disebut juga NADH-Q oksidoreduktase
Suatu multi subunit besar, berbentuk L
Berfungsi memindahkan elektron dari NADH ke koenzim Q dikopel
dgn transfer 4 ion H+ melintasi membran dalam

NADH + Q + 5H+ matriks NAD + QH + 4H+

Pada komplek I awalnya terjadi transfer elektron dari NADH
(hasil katabolisme nutrien) ke FMN (dlm kompleks).

Kemudian elektron berpindah dari FMN ke pusat Fe-S (serial).

Selanjutnya elektron berpindah dari Fe-S ke koenzim Q.

Kompleks II:

(Suksinat-Q reduktase)
FADH2 hasil siklus Krebs dan elektron ditransfer melalui
beberapa pusat Fe-S ke Quinone

Gliseraldehid 3-P yg berasal dari pemecahan Trigliserida (TG)

atau dari glikolisis dan asetil koA mentransfer elektron ke
Quinone melalui jalur lainnya
Respiratory Control

1. Mainly by availability of ADP

(ATP/ADP transporters may be rate limiting
at certain times)
2. Availability of electrons
NADH/NAD ratio or FADH2/FAD
3.Availability of O2
The respiratory chain can be
inhibited by:

a. Inhibitors of the respiratory chain,

b. Inhibitors of oxidative phosphorylation,
c. and uncouplers of oxidative phosphorylation.
Inhibitor of respiratory chain
Rotenone is a common insecticide that strongly inhibits the
electron transport of complex I. Rotenone is a natural
product obtained from the roots of several species of
plants.

Tribes in certain parts of the world beat the roots of trees

along riverbanks to release rotenone into the water which
paralyzes fish and makes them easy prey.
 Amytal is a barbiturate that inhibits the electron
transport of complex I.
Demerol is painkiller that also inhibits complex I.

All three of the complex I inhibitors block the

oxidation of Fe-S clusters of complex I
 2-Thenoylfluoroacetone and carboxin
specifically block electron transport in
Complex II

Malonate is a competitive inhibitor of

Complex II.
 Antimycin A and dimercaprol inhibit the respiratory chain
at Complex III.
Antimycin A1 is an antibiotic that inhibits electron
transfer in complex III by blocking the transfer of
electrons between Cyt bH and coenzyme Q bound at the
QN site.
 Cyanide, azide and carbon monoxide all
inhibit electron transport in Complex IV. All
of the inhibitors, inhibit electron transfer by
binding tightly with the iron coordinated in
Cyt a3.
Carbon monoxide is toxic due to its affinity for the
heme iron of hemoglobin. Animals carry many
molecules of hemoglobin, therefore it takes a large
quantity of carbon monoxide poisoning. Animals
have relatively few molecules of Cyt a3.
Consequently an exposure to a small quantity of
azide or cyanide can be lethal. The toxicity of
cyanide is solely from its ability to arrest electron
transport.
Inhibitors of oxidative
phosphorylation
Atractyloside inhibits oxidative phosphorylation
by inhibiting the transporter of ADP into and ATP
out of the mitochondrion

The antibiotic oligomycin completely blocks

oxidation and phosphorylation by blocking the
flow of protons through ATP synthase
Uncouplers of oxidative
phosphorylation
Uncouplers dissociate oxidation in the respiratory
chain from phosphorylation

The uncoupling of oxidative phophorylation from

electron transport generates heat.

Thermogenin (or the uncoupling protein) is a

physiological uncoupler found in brown adipose tissue
that functions to generate body heat, particularly for the
newborn and during hibernation in animals.
Inhibition sites of the respiratory chain
 Mitochondrial Abnormalities
The condition known as fatal infantile
mitochondrial myopathy and renal
dysfunction involves severe diminution or
absence of most oxidoreductases of the
respiratory chain.

MELAS (mitochondrial encephalopathy, lactic

acidosis, and stroke) is an inherited condition
due to NADH-Q oxidoreductase (Complex I) or
cytochrome oxidase (Complex IV) deficiency
 Link Video:
Cellular Respiration:
https://www.youtube.com/watch?v=Fcu_8URp
4Ac

ATP Synthase:
https://www.youtube.com/watch?v=PjdPTY1w
HdQ