ELECTRON TRANSPORT

AND CLINICAL
CORRELATION
ADVANCE BIOCHEMISTRY
OUTLINE
MITOCHONDRIA
ELECTRON TRANSPORT CHAIN
CLINICAL CORRELATIONS
MITOCHONDRIAL MEMBRANE:
Structure and Compartmentation
Mitochondria
Site of oxidative phosphorylation
Powerhouse of the cell
Number and activity reflects the role of tissue in
aerobic metabolic functions and its need for energy
Example is the Cardiac Muscle
of the
cytoplasmic
membrane consist
of mitochondria
Inner membrane
contain numerous
invaginations
(cristae)
MITOCHONDRIA
- Shapes depend on cell type
Oblong or cylindrical
spherical
Cardiac cells
Liver
- Hepatocyte
contain 800-
2000
mitochondria
RBC- no
mitochondria
and obtain
energy from
glycolysis


Intermembrane space
OUTER MEMBRANE INTERMEMBRANE SPACE INNER MEMBRANE
Rich in integral protein
(porin or VDAC, voltage
dependent anion
channel
Monoamine oxidase &
Kynurenine hydroxylase
for removal of
neurotransmitter
Enzymes involve in the
trasnfer of energy
80 % protein
Rich in unsaturated fatty
acids
Cardiolipin is present in
high concentrations
Enzyme complexes and
oxidative
phosphorylation
MATRIX
mtDNA
Ribosomes
Proteins for transcription
Enzymes of the TCA except succinate dehydrogenase
ELECTRON TRANSPORT CHAIN
NADH and FADH
2
are oxidized by the ELECTRON
TRANSPORT CHAIN
Reducing equivalents are converted to utilizable
energy as ATP
Done by Oxidative Phosphorylation
OXIDATION- REDUCTION REACTIONS
Mitochondrial electron transport consists of a
sequence of linked oxidation- reduction reactions

Transfer electrons from a suitable electron donor (
reductant) to a suitable electron acceptor (oxidant)

REDOX COUPLE or PAIR- an oxidant and its reductant


OXIDATION- REDUCTION REACTIONS
Oxidation- Reduction Potential
The ease with which an electron donor (reductant)
gives up its electron to an electron acceptor (oxidant)
Measured in volts as electromotive force (emf) of a
half cell made up of an oxidation-reduction couple
when compared to a standard reference half cell
hydrogen electrode reaction
OXIDATION- REDUCTION REACTIONS
Standard hydrogen electrode is set at 0.0 V at pH 0.0

At pH 7.0, the reference hydrogen potential is 0.42
V
MULTICOMPONENT SYSTEM
Electron transport, also known as aerobic
respiration, is the last stage of aerobic metabolism
After glycolysis and TCA cycle, 10 NADH and 2 FADH2
are generated from the oxidation of one glucose
molecule.
When there is sufficient O2 supply, NADH and FADH2
enter electron transport chain to become reoxidized



MULTICOMPONENT SYSTEM
Large amount of energy is recovered, when
electrons are passed from NADH and FADH2 to O2.
This is accomplished by a series of carrier protein in
the inner mitochondrial membrane
Not organized in linear arrangement but are grouped
into four large complexes
Electron Transport and Oxidative
Phosphorylation
In ETS, electrons are transferred from NADH and
FADH2 to O2 step by step.

In the mean time, energy released from electron
flow is coupled to ATP synthesis.

Here, phosphorylation of ADP is coupled with the
oxidation of NADH or FADH2.


Composition of the Electron Transport Chain
Large protein complexes.

Complex I - NADH-ubiquinone oxidoreductase
Complex II - Succinate-Ubiquinone oxidoreductase
Complex III Ubiquinol -Cytochrome c reductase
Complex IV - Cytochrome c oxidase
Complex V- ATP synthase



Complex I - NADH-ubiquinone oxidoreductase

Entry Point of NADH
Transfers electrons from NADH to obiquinone (co-
enzyme Q) coupled with the transport of four
protons across the membrane required for the
synthesis of ATP
CoQ ubiquinone

Highlighted region serves as an anchor to inner
mitochondrial membrane.

Reduction of CoQ

Complex I - NADH-ubiquinone oxidoreductase

FMN (Flavin Mononucleotide- located in different
subunits of the hydrophilic arm of complex I
NADH + H
+
+ FMN ------> NAD
+
+ FMNH
2
Series of FeS center (2 Fe2 S and 4Fe S type)
reduce ubiquinone to ubiquinol

4 protons are translocated across the inner
membrane to the intermembrane space

Complex II - Succinate-Ubiquinone
oxidoreductase

Known as Succinate dehydrogenase
Entry point for FADH
2

Succinate dehydrogenase (from the citric acid cycle)
directs transfer of electrons from succinate to CoQ
via FADH
2
.

Acyl-CoA dehydrogenase (from -oxidation of fatty
acids) also transfers electrons to CoQ via FADH
2
.
Succinate- fumarate + 2 H
+
+ 2 e


All electrons from FADH2 and NADH
must pass through CoQ.

Known as cytochrome bc1 complex
Catalyzes transfer of 2 electrons from ubiquinol to
cytochrome c with translocation of four protons
across the membrane.
Consist of 3 prosthetic groups that serve as redox
center
A. Cytochrome b
B. Cytochrome c1
C. Rieske iron sulfur proteins
Complex III Ubiquinol -Cytochrome
c reductase
CYTOCHROMES
Proteins that contain a heme group tightly bound
Iron in heme of a cytochrome is alternately oxidized (
Fe
3+)
or reduced ( Fe
2+)
as it functions in the transport
of electrons
Designated as a, b, and c on the basis of alpha band
of their absorption spectrum and the type of heme
group attached to the protein

Complex III Ubiquinol -
Cytochrome c reductase
Cytochrome B
Heme b562
Heme 566
Cytochrome c1
1 heme group
Rieske- iron sulfur protein
2 Fe2S clusters
Complex III Ubiquinol -
Cytochrome c reductase
Cytochrome c is a
mobile electron carrier
Bind to cytochrome c1
of complex III and accept
electron
Heme iron is
coordinated to a
nitrogen of histidine and
a sulfur iron of a
methionine

Complex III Ubiquinol -
Cytochrome c reductase

Complex IV - Cytochrome c oxidase

Components of ATP synthase

These are knob-like projections
into the matrix side of the inner
membrane.

Two units
F
1
contains the catalytic site for
ATP synthesis.
F
0
serves as a transmembrane
channel for H
+
flow.

F
1
-F
0
complex serves as the
molecular apparatus for coupling
H
+
movement to ATP synthase.
ATP is transported from the matrix of mitochondria
to cytosole by ATP-ADP translocase.

ATP and ADP cannot diffuse through the mitochondria
membrane freely.

The exit of ATP is coupled with the entry of ADP
into mitochondria.
Regulation of oxidative phosphorylation
Electrons do not flow unless ADP is present for
phosphorylation
Increased ADP levels cause an increase in the activity
of various enzymes including:

glycogen phosphorylase
phosphofructokinase
citrate synthase
ATP is transported from the matrix of mitochondria
to cytosole by ATP-ADP translocase.

ATP and ADP cannot diffuse through the mitochondria
membrane freely.

The exit of ATP is coupled with the entry of ADP
into mitochondria
Regulation of oxidative phosphorylation
Electrons do not flow unless ADP is present for
phosphorylation
Increased ADP levels cause an increase in the activity
of various enzymes including:

glycogen phosphorylase
phosphofructokinase
citrate synthase

Recycling of cytoplasmic NADH(?)
Different methods are used to recycle NADH. This
accounts for the different energy productions from
glucose.

Glycerol-3-phosphate shuttle
Used by skeletal muscles and the brain

Malate-aspartate shuttle
Used by the heart and liver

Glucose-3-phosphate shuttle
Malate-aspartate shuttle
MITOCHONDRIAL GENES AND DISEASES
Contain their own genome
Contains 13 proteins of
electron transport chain
Not self replicating
organelles

90% mitochondrail proteins are
encoded in nuclear DNA
Synthesized in the cytosol
Imported in the mitochondria

MITOCHONDRIAL GENES AND DISEASES
MITOCHONDRIAL DEFECTS
Degenerative disease of aging (Parkinson and
Alzhiemer Disease)
Point mutation in mtDNA
Deletion of large portions of mtDNA
DECREASED ACTIVITY OF THE ELECTRON TRANSPORT
CHAIN which leads to the accumulation of pyruvate and
fatty acids= lactic acidosis and triglyceride accumulation
Complex I
Mutations in subunits have
been shown to lead to a
number of nuerode
degenerative disease.
Source of reactive oxygen
species damage mitochondrial
DNA and may be cause of aging.
Leber Hereditary Optic Neuropathy( OMIM
53500)
Maternally inherited
Affects the nervous system
Including optic nerves causing sudden onset
blindness in early adulthood due to the death of
optic nerve
Results from single base changes in the
mitochondrial genes encoding 3 subunits o complex I
(ND1, ND4, and ND6)
Lower the activity o complex 1
MITOCHONDRIAL MYOPATHIES FROM
MUTATIONS IN MITOCHONDRIAL tRNA genes
Mutation in the tRNA gene for lysine
- myclonic epilepsy and ragged red fibers (
MERRF) ( OMIM 545000)
Symptoms: myoclonus and ataxia with generalized
seizurees and myopathy

Skeletal muscle contain abnormally shaped
mitochnodria that contain paracrystalline structures
giving an appearance of ragged red fibers

Mutation in the tRNA for leucine
- common mitochondrial encephalopathy
- lactic acidosis
- stroke like activity
( MELAS) (OMIM 540 000)
- Skeletal muscles contain ragged red fibers but retain
cytochrome c oxidase activity
MITOCHONDRIAL MYOPATHIES FROM
MUTATIONS IN MITOCHONDRIAL tRNA genes