1

OXYGEN RADICALS
and AGING
Rondang R. Soegianto
2009
2
Oxidative Damage and Aging are

two processes commonly found

in eukaryotic organisms
3
ENERGY, essential for life processes

In humans,

Electron Transport System (ETS) in

mitochondria is main mechanism for

aerobic energy supply
4
ATP = Energy currency of the cell

Produced in mitochondria via

Oxidative Phosphorylation (OXPHOS)

Oxidation processes in living systems are
catalyzed by class I enzymes:

OXIDOREDUCTASES
5

Oxidoreductases (Harper 26th)



1. Oxidases: A Containing Cu
B As flavoproteins

2. Dehydrogenases:
A. NAD+ or NADP+ as coenzyme
B Flavin as coenzyme
C Cytochromes (Fe-porphyrin as coenzyme)

3. Hydroperoxidases:
A Peroxidase
B Catalase

4. Oxygenases:
A Dioxigenase
B Monooxigenase
6
1. Oxidases:
- Remove 2 protons (H+) from substrate and pass to oksigen
- Generate H
2
O or H
2
O
2

Two groups of oxidases:
A Containing Cu
Example: Cytochrome a3 (cyt a3) also known as cyt aa3
Is a cytochrome oxidase
Terminal compound of the respiratory chain in
mitochondria

B. Flavoproteins, contain FMN or FAD
Ex. : L-aminoacid oxidase
Xanthine oxidase
Aldehyde dehydrogenase
7
3. Hydroperoxidases use H
2
O
2
as substrate

A. Peroxidase reduces peroxides using various e-
acceptors

H
2
O
2
+ AH
2
2 H
2
O + A

In erythrocytes and other tissues:

H
2
O
2
+ 2 GSH GSSG + H2O

GSH = Reduced gluthatione
Glutamyl-cysteinyl-glycine (a tripeptide)
-SH = Reducing group of cysteine residue

Peroxidase
Gluthatione peroxidase
8
B. Catalase
Hemoprotein with 4 heme groups

2 H
2
O
2
2 H
2
O + O
2


Found in: Blood, bone marrow, mucous
membranes
Kidney, liver

Catalase destroys peroxides formed by oxidases

Catalase
9
Free radicals

Transfer of a single e to O O (superoxide anion)
Can damage membranes, DNA, etc.

Destructive effects
Amplified by: Free radical chain reaction
Removed by: Superoxide dismutase (SOD) in the reactions

O + O 2H H O + O

H O 2H O + O

Catalase
2 2 2
-
2
-
2
+
2
2
SOD
2 2
2
-


2
2
-
10
Mitochondria
Make > 90% of cellular ATP
Powerplant of the cell
Four Compartments
Matrix has numerous enzymes that reduce
NAD
+
to NADH during catabolism of foodstuffs
Inner Membrane has:
Proteins that transfer electrons (the ETS)
ATP synthase
Intermembrane Space
Outer Membrane
11
Faces of mitochondrial membrane (V & V Fig. 20-3)

12


Role of RC of mitochondria in the conversion of food energy to ATP
Harper 26 Fig. 12-2
13
Harper 26 Fig. 12-4
14
15
16
17
18
19
20
Reactive Oxygen Species (ROS) are
oxygen radicals that may cause damage
to cells and tissues, such as in
- Neurodegenerative diseases
(Alzheimer, Parkinson)

-Cardiovascular diseases

- Rheumatoid Arthritis (RA)
(Bone erosion, cartilage loss,
loss of joint function)
21
In RA :

ROS does not destroy collagen directly

ROS induces synthesis of
Matrix Metalloproteinase (MMP)

that attacks connective tissues
Therapy targeted at possible MMP inhibitors
22
Effect of ROS on
a. Lipid and Nucleic Acids
Double bonds, easy target for oxidative
damage (lipid peroxidation)

b. Cellular protein and Enzymes
Oxidation produces altered proteins
23
Altered proteins accumulate with aging.

This will interfere with normal homeostasis

Can cause age related pathologies, such as
- Atherosclerosis
- Senile cataract
- Diabetes Mellitus
- Immune system failure
- Neurodegenerative diseases
24
Age related changes in enzymes
and other proteins:

- Alteration in catalytic activities
- Altered folding in the 3-D structure

The products are altered (abnormal) proteins
These have to be eliminated (degradation)

With aging: Disturbed balance between
accumulation and degradation of
modified forms.
25
Other type of age associated modification:

Protein Glycosylation
(non enzymatic)

Causes aging of long lived proteins
Such as: Collagen
Crystallin
26
Glycosylation = Maillard reaction

Addition of carbohydrate to protein
Normally for protein secreted by cell
or protein bound to cell surface

Hb can be glycosylated in blood glucose
27
Inactivation by glycosylation occurs
in proteins with lysine in critical location

Superoxidismutase
Ribonuclease
Na+, K+-ATPase

Experimentally inactivated when
incubated in glucose
28
Final products of glycosylaton called

Advanced Glycosylation Endproducts (AGE)

Free radicals induce formation and
accumulation of AGE

ANTIOXIDANTS inhibit protein glycation
And accumulation of AGE
29
With increasing age and glucose
concentration

accumulation of AGE in plasma
and vessel walls

cause many diabetic complications
(cataract, atherosclerosis)
30
Other type of protein modification due
to oxidative stress is

formation of protein-protein Cross Links

caused mainly by disulfide bonds
31
Mitochondrial Dysfunction

ROS generated during aging
Chronic oxidative damage to Electron
Transport System (ETS)

Resulting decrease in functional capacity
of cells and tissues during aging
32
Caloric Restriction (CR)

Since major endogenous source of ROS
is mitochondrial respiration,

CR markedly reduces production of
Superoxide and H
2
O
2

Effect of CR most striking in brain
33
Kristal and Yu (1992)

Age related deterioration is produced

by the sum of the damage induced by

free radicals, by glycation (Maillard reaction)
and by their interactions.
34
Reference:

OXYGEN RADICALS and the

DISEASE PROCESS

Thomas, C.E., Kalyanaraman, B. eds

Harwood academic publishers