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Oxygen

A double-edged sword

ROS/AOS
Reactive Oxygen Species (ROS) / Active Oxygen Species (AOS) Molecular oxygen is continually reduced within cells to several forms of Reactive Oxygen Species Produced during
Photosynthesis and respiration By-products of metabolism Dedicated enzymes

ROS-generating processes in chloroplasts, mitochondria, and peroxisomes Oxidation targets include proteins, DNA, and lipids

Basics of Redox Chemistry


Oxidation
Gain in oxygen Loss of hydrogen Loss of electrons

Reduction
Loss of oxygen Gain of hydrogen Gain of electrons

Oxidant
Oxidizes another chemical by taking electrons, hydrogen, or by adding oxygen

Reductant
Reduces another chemical by supplying electrons, hydrogen, or by removing oxygen

Radical and Non-Radicals


Free Radicals :
Any species capable of independent existence that contains one or more unpaired electrons A molecule with an unpaired electron in an outer valence shell

Non-Radicals:
Species that have strong oxidizing potential Species that favor the formation of strong oxidants (e.g., transition metals, H2O2)

Reactive oxygen species (ROS)

Oxygen is normally reduced by four electrons to produce water Reaction catalysed by complex IV (cytochrome oxidase) of the mitochondrial electron transport chain Single electron transfer to molecular oxygen leads to ROS
Biologically important oxygen species
Superoxide radical anion Singlet Oxygen Hydrogen Peroxide Hydroxyl radicle

Inter conversion of active oxygen species

Light
e1O 2

O2 SOD

Cu, Zn (Chloroplast/ Cytosol) Fe (Chloroplast) Mn (Mitochondria)

H2O2 APX

1 2

Chloroplast Cytosolic Glyoxysome

3 4

H2O

SOD types
Cu and ZnSOD
dimmer with subunit 32kDand each with one Cu2+ and Zn+mainly distributes in higher plant chloroplast mainly distributes in prokaryotic bacteria as well in eucaryotic mitochondria is a basic typemainly locates in green alga.

MnSOD

FeSOD

SOD catalytic reaction


2 O2
+2H2O SOD H2O2+O2

Production of Hydroxyl radical (OH)

Transition metal catalyzed Other reductants can can suppliment Fe2+ like GSH, ascorbate, hydroquinones) Fe2+ is an extremely reactive oxidant

Redox balance
Cells maintain a particular redox balance. Antioxidant activity monitors ROS levels. The electronegativity of the cell is maintained by : reduced glutathione (GSH)

ROS generation
From electron transport and enzymatic sources (Chloroplast & Mitochondria) Glycollate oxidase
Photorespiration

Acyl CoA oxidase


oxidation of lipids, in peroxisomes

NADPH oxidase (rbohA ) Oxidative Burst : plants

Role of H2O2

Oxidative stress and cell damage


High doses:
Directly damage/kill cells/ HR

Low doses/chronic overproduction of oxidants:


activation of cellular pathways stimulation of cell proliferation damage to cellular proteins, DNA and lipids

Consequences of lipid peroxidation

Structural changes in membranes


alter fluidity and channels alter membrane-bound signaling proteins increases ion permeability

Lipid peroxidation products form adducts/crosslinks with non lipids


e.g., proteins and DNA

Cause direct toxicity of lipid peroxidation products


e.g., 4-hydroxynonenal toxicity

Disruptions in membrane-dependent signaling DNA damage and mutagenesis

Consequences of protein oxidation

Oxidation of catalytic sites on proteins


loss of function/abnormal function

Formation of mixed sulfide bonds


Protein-protein linkages (RS-SR) Protein-GSH linkages (RS-SG)

Alteration in 2 and 3 structure Increased susceptibility to proteolysis

Consequences of protein oxidation

DNA adducts/AP sites/Strand breaks


Mutations initiation of cancer

Stimulation of DNA repair


can deplete energy reserves (PARP) imbalanced induction of DNA repair enzymes induction of error prone polymerases activation of other signaling pathways

DNA oxidation products

Oxidation of deoxyribose (DNA backbone)

Mitochondria as a source of ROS

Productions of 1O2 in PSII and of O2- in PSI in chloroplast thylakoids

Peroxisomes as a source of ROS

Beta Oxidation

Enzymes in mammalian peroxisomes that generate ROS

ROS from Chloroplast

The reduced ferredoxin (Fd) is a major source of Superoxide radical Interior of thylakoid membranes (PETC)

Cellular antioxidants

Small Molecules Water soluble: glutathione, uric acid, ascorbate (Vit. C) Lipid soluble: -tocopherol (Vit. E), -carotene, coenzyme Q Proteins Intracellular: SOD (I and II), glutathione peroxidase, catalase Cell membrane: SOD (III), GPx, plasma proteins (e.g. albumin) Extracellular: phospholipid hydroperoxide GPx (PHGPx)

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