ROS and Free Radical Injury:

Learning Objectives:
1. Define: radical and the different types
a. Radical- molecules that have a single unpaired electron
usually in their outer orbital. They are highly reactive
and initiate chain reactions. How? By extracting an
electron from a neighboring molecule to complete their
own outer orbitals, thereby, initiating free radical chain
reactions.
b. Types of radicals:
Bound radicals exits in the active site of
enzymes or as part of the functional property of
a protein, e.g. Fe in hemoglobin & Cu in catalase
Free radicals -- exists independently, i.e. are
not associated with an other molecule such as
in the active site of an enzyme.
Antioxidants are stable free radicals
Vits E and C accept electrons (antioxidant
activity) to form stable free radicals that
distribute the free electrons over their
molecular ring structure.
Reactive Oxygen Species (ROS) are free
radicals of oxygen: superoxide, hydrogen
peroxide & hydroxyl radical.
Transition metals (Fe, Cu, Mo) -- are free
radicals when not normally bound to proteins
that tend to stabilize them.
2. Describe reduction of oxygen by four one-electron steps:
a. Oxygen accepts:
1 e- = Superoxide
2 e- = hydrogen peroxide: H2O2
o REMEMBER- its not a radical
3 e-= hydroxyl radical
4 e- = gets reduced to water
3. List the 5 major ROS: VERY IMPORTANT
a. Superoxide (O2-)
Water soluble with limited lipid solubility
Produced by coenzyme Q in the ETC
b. Hydrogen peroxide (H2O2)
Lipid soluble
Not a free radical, but a precursor of
hypochlorous acid (HOCl) produce in
neutrophiles
*Generated in peroxisomes during lipid
metabolism
c. Hydroxyl radical (HO)
 Water soluble
The *most reactive free radical* produced from
H2O2 reacting with Superoxide or transition metals,
e.g. Fe or Cu
d. Organic radical (e.g. ROO)
Lipid soluble
Generated by lipid peroxidation, free radical attach
on unsaturated double bonds found in membrane
lipids
e. Reactive nitrogen-oxygen species (RNOS)
Water soluble
Derived from free radical nitric oxide (NO)

4. Identify the reactants involved in the production of the

hydroxyl ion.:
a. H2O2 reacts with superoxide and/or transition metals like Fe
and Cu to produce the hydroxyl radical.

5. State major sources of primary ROS

a. Identify the reactants involved in the production of the
hydroxyl ion.:
CoQ -- generates superoxide
b. Oxidases --degrade neurotransmitters
 Monoamine oxidase
Dopamine oxidase
c. Oxygenase liver detoxification
d. Cytochrome P450 (monooxygenase)
Dextoxifications of alcohol, drugs, chemical
toxicants (CCL4)
Lipoxygenase in leukotrienes synthesis
e. Peroxidases
GSH peroxidase in prostaglandin synthesis
f. Ionizing radiation
Split water into hydroxyl radical and hydrogen
radicals
Generate organic radicals
Skin cancer & cell death
6. Describe the mechanisms of cellular damage by oxygen
radicals.
a. Overview:

b.

7. Discuss cellular effects of NO at lower and higher

concentration.
a. Attack Membrane Lipids
b. Form lipid peroxide and lipid peroxy radicals
c. Defended by vitamin E and lipid soluble antioxidants
d. Attack Cell Proteins and Peptides
e. Cause fragmentation ,
f. cross linking and aggregation
 g. Defended by water soluble
h. glutathione and phagocytic cells
i. Attack DNA
j. Nitrogenous bases alterations,
k. such that A-G base pairs occur.
l. Breaks in the deoxyribose backbone
m. Defended by DNA repair enzymes
n.
8. Discuss cellular effects of RNOS.
9. State the functions of the enzymes involved in respiratory
burst.
10. Describe the function of NADPH.
11. List all free radical scavengers and state their functions
12. Describe the Anti-Oxidant/Pro-Oxidant Paradox