ROS REVIEW ARTICLES

Superoxide in Biology and Medicine: An Overview

Robert Z. Hopkins

AIMSCI Research Institute, P.O. Box 37504, Raleigh, NC 27626, USA

Correspondence: [email protected] (R.Z.H.)

Hopkins RZ. Reactive Oxygen Species 1(2):99–109, 2016; ©2016 Cell Med Press
http://dx.doi.org/10.20455/ros.2016.825
(Received: January 12, 2016; Revised: January 31, 2016; Accepted: February 1, 2016)

ABSTRACT | Since 1933 when Linus Pauling, a twice-honored Nobel laureate, proposed its existence based
on the theory of quantum mechanics, superoxide has gradually taken central stage in the research field of
reactive oxygen species (ROS) in biology and medicine. Indeed, superoxide is considered the primary ROS
that gives rise to secondary ROS. This oxygen free radical is generated in a wide variety of biological systems
ranging from aerobic microorganisms to human cells, and also formed in the deep ocean and the soils of Earth
and possibly the soils of Mars as well. Superoxide is now recognized as an important molecule that is formed
via defined mechanisms and involved in diverse physiological and pathophysiological processes. This article
provides an overview on the basic chemistry and biochemistry of this ubiquitous oxygen free radical and its
significance in biology and medicine.

KEYWORDS | Cytochrome P450 system; Fenton reaction; Haber-Weiss reaction; Mitochondria; NADPH
oxidase; Reactive oxygen species; Redox cycling; Redox signaling; Superoxide; Superoxide dismutase;
Uncoupled endothelial nitric oxide synthase; Xanthine oxidoreductase

ABBREVIATIONS | Cu,ZnSOD, Cu,Zn superoxide dismutase; CYP, cytochrome P450; ECSOD, extracellu-
lar superoxide dismutase; EPR, electron paramagnetic resonance; METC, mitochondrial electron transport
chain; MnSOD, manganese superoxide dismutase; NOX, NAD(P)H oxidase; ROS, reactive oxygen species;
SOD, superoxide dismutase; XO, xanthine oxidase

CONTENTS

1. Introduction
2. Sources of Superoxide
3. Chemistry and Biochemistry of Superoxide
3.1. Oxidation of Iron-Sulfur Clusters
3.2. Reaction with Nitric Oxide
3.3. Haber-Weiss Reaction and Fenton Reaction
3.4. Membrane Permeability and Protonation
4. Superoxide in Biology and Medicine
4.1. Phagocyte-Mediated Immunity
4.2. Redox Modulation and Signaling

REACTIVE OXYGEN SPECIES ǀ aimsci.com/ros 99 VOLUME 1 ǀ ISSUE 2 ǀ MARCH 2016

 ROS REVIEW ARTICLES
4.3. Autoregulation of Superoxide Production
4.4. Cell and Tissue Defenses
4.5. Pathophysiology
5. Perspectives

1. INTRODUCTION

Superoxide, also known as superoxide anion or su-

peroxide anion radical, is designated as O2˙ˉ, where
the dot denotes the unpaired electron and the minus
sign denotes that the species is negatively charged.
The above three names for O2˙ˉ are used inter-
changeably in the literature, and for simplicity, su-
peroxide is used throughout this article. Superoxide
FIGURE 1. Electron configuration illustration of
can exist in a protonated form, known as perhydroxyl
one electron reduction of molecular oxygen to su-
or hydroperoxyl radical (HO2˙) with a pKa of 4.8.
peroxide. Molecular oxygen (O2) is in fact a di-
Because of this pKa, superoxide exists predominant-
radical because it contains two unpaired electrons.
ly in the non-protonated form (O2˙ˉ) under physio-
The same spin direction of the two unpaired elec-
logical pH.
trons in O2 causes spin restriction, making O2 less
Linus Pauling, a twice-honored Nobel laureate
reactive. On the other hand, superoxide (O2˙ˉ) con-
(Chemistry in 1954; Peace in 1962) proposed the ex-
tains one unpaired electron and has one more elec-
istence of superoxide based on the theory of quantum
tron than O2, and as such, superoxide is negatively
mechanics in 1933 (see in ref. [1]). Two decades af-
charged. Superoxide can become protonated to form
ter Linus Pauling’s proposal, in 1954, Rebecca
perhydroxyl radical (HO2˙.), which is believed to be
Gershman and associates suggested that superoxide
more reactive than O2˙ˉ.
might be responsible for both oxygen toxicity and
the deleterious effects of x-irradiation [2]. Subse-
quently, xanthine oxidase was proposed to produce
superoxide in 1962 [3], and this was proved by elec- e- → O2˙ˉ [7] (Figure 1). In biological systems, su-
tron paramagnetic resonance (EPR) studies in 1969 peroxide is generated from various metabolic pro-
[4]. Meanwhile, methods involving electrolytic uni- cesses. These include the mitochondrial electron
valent reduction of molecular oxygen were used to transport chain (METC) complexes, NADPH oxi-
produce superoxide. With superoxide generated via dases (also known as NOXs), xanthine oxidoreduc-
the above approaches, Joe McCord and Irvine Fri- tase, uncoupling of endothelial nitric oxide synthase
dovich discovered Cu,Zn superoxide dismutase (eNOS), and cytochrome P450 enzyme system,
(Cu,ZnSOD) in 1969 [5]. Subsequently, Bernard among others (e.g., Rac,) (Figure 2).
Babior and coworkers reported superoxide genera- Both NOXs and mitochondria are key machineries
tion by respiratory burst (activation of NADPH oxi- in cellular superoxide generation. Mitochondria are
dase) of leukocytes as a potential mechanism of generally considered the chief cellular source of su-
bactericidal activity [6]. Now superoxide is consid- peroxide, whose formation is primarily derived from
ered one of the most important reactive oxygen spe- the univalent reduction of molecular oxygen by elec-
cies (ROS) that plays a significant role in a variety of trons leaked from the METC [8]. This is not surpris-
biological processes and disease conditions. ing as mitochondrial respiration accounts for ~90%
of total cellular oxygen consumption. Recently,
NOX4 is also found to be present in mitochondria
2. SOURCES OF SUPEROXIDE and acts as a potential source of mitochondria-
derived superoxide [9, 10]. Hence, mitochondria-
Superoxide is produced from one electron reduction derived superoxide is not necessarily always due to
of molecular dioxygen (oxygen for simplicity): O2 + electron leakage from the METC. In addition to its

REACTIVE OXYGEN SPECIES ǀ aimsci.com/ros 100 VOLUME 1 ǀ ISSUE 2 ǀ MARCH 2016

 ROS REVIEW ARTICLES

FIGURE 2. Major sources of cellular superoxide. NADPH oxidases (NOXs) and mitochondria are consid-
ered the major cellular sources of superoxide. Many other enzymes and cellular processes also contribute to
cellular superoxide production, such as xanthine oxidoreductase, cytochrome P450 (CYP) enzyme system (es-
pecially CYP2E1), and uncoupled endothelial nitric oxide synthase (eNOS).

presence in mitochondria, NOX4 is also found to un- implicated in a number of pathophysiological pro-
dergo translocation into the nucleus via a leukotriene cesses, such as atherosclerosis, inflammation, and
C4-dependent mechanism, and its presence in the tissue ischemia-reperfusion injury in the heart, liver,
nucleus contributes to oxidative damage to the nu- and kidneys [12].
clear DNA under stress conditions, including in- Cytochrome P450 (CYP) enzyme system, especial-
flammatory responses and treatment with cancer ly CYP2E1, is also a significant source of superoxide
chemotherapeutic agents [11] formation in cells, particularly under conditions
Notably, xanthine oxidoreductase has two inter- when these enzymes are induced by chemicals (e.g.,
convertible forms, namely, xanthine dehydrogenase ethanol). CYP2E1-derived superoxide/ROS upon
and xanthine oxidase (XO). Both forms catalyze the ethanol consumption may also be responsible for the
conversion of hypoxanthine to xanthine, and xan- upregulation of CYP2A5/2A6 by alcohol [13]. Re-
thine to uric acid. Both forms also catalyze one- and cently, a redox signaling heme-containing globin,
two-electron reduction of molecular oxygen to form namely, GLB-12, in Caenorhabditis elegans has
superoxide and hydrogen peroxide (H2O2), respec- been shown to generate superoxide, and the superox-
tively, with XO being more active in generating the ide produced may play a critical signaling role in
above ROS. The reversible conversion of xanthine regulating apoptosis and controlling the reproduction
dehydrogenase to XO occurs under certain condi- of the organism [14].
tions including hypoxia/tissue ischemia and oxida- In addition to the aforementioned endogenous
tion of the cysteine thiol groups of the enzyme. sources, superoxide is also formed during biotrans-
Xanthine oxidoreductase-derived ROS have been formation of certain xenobiotics, especially quinone

REACTIVE OXYGEN SPECIES ǀ aimsci.com/ros 101 VOLUME 1 ǀ ISSUE 2 ǀ MARCH 2016

 ROS REVIEW ARTICLES
parent absence of organic material at the Martian
surface [18].

3. CHEMISTRY AND BIOCHEMISTRY OF

SUPEROXIDE

Although it is called “super-oxide”, superoxide is in

fact not a strong oxidizing species. Because of its
lower reduction potential (–330 mV for the O2/O2˙ˉ
redox couple), superoxide often acts as a reducing
FIGURE 3. Schematic illustration of the concept agent rather than an oxidizing species. Indeed, the
of chemical redox cycling. Redox cycling of qui- ferricytochrome c reduction assay, a method for de-
none compounds gives rise to an enormous amount tecting superoxide, is based on the ability of superox-
of superoxide via one electron reduction of molecu- ide to reduce this small heme-containing protein [19].
lar oxygen by the semiquinone radical intermediate. The ability of superoxide to directly oxidize biomol-
ecules, including lipids, proteins, and nucleic acids is
much limited. However, superoxide is an important
compounds, via a process known as redox cycling ROS that can result in cell and tissue injury [20]. As
(Figure 3). Chemical redox cycling may be defined outlined below, the biologically damaging potential
as the reduction and oxidation cycle between two of superoxide is attributed to its several unique
forms of a compound, frequently a quinone molecule. chemical and biochemical properties (Figure 4).
Redox cycling is typically initiated by a univalent
reduction of a quinone molecule to form a semiqui- 3.1. Oxidation of Iron-Sulfur Clusters
none radical species. The CYP enzyme system and
METC are among the major machineries that carry Although it generally acts as a reducing agent, su-
out the one electron reduction. The semiquinone rad- peroxide does oxidize the iron-sulfur clusters in sev-
ical formed can then donate one electron to molecu- eral enzymes, including aconitase, an enzyme of the
lar oxygen, and during this reaction, molecular tricarboxylic acid cycle [21, 22], and the mitochon-
oxygen is reduced to a superoxide anion radical, and drial electron transport chain enzyme complexes [23].
the semiquinone radical is oxidized back to the orig- Oxidation of the iron-sulfur clusters by superoxide in
inal quinone molecule. Hence, the entire process in- these enzymes leads to the release of iron from the
volving one electron reduction and oxidation forms a enzymes and the enzyme inactivation. The released
cycle, leading to the persistent production of super- iron ions may participate in the Fenton reaction, re-
oxide and secondary ROS (e.g., hydrogen peroxide sulting in the formation of hydroxyl radicals (see
and hydroxyl radicals). Section 3.3 below). In contrast, reversible oxidation
In the sea, superoxide is generated by photolysis of of the iron-sulfur cluster in prokaryotic transcription
organic matter in the photic zone as well as bacteria factors SoxR/SoxS leads to their activation (see Sec-
in the deep ocean with the latter being a vast source tion 4.2 below).
of superoxide [15, 16]. Superoxide is formed in de-
sert soils via photochemical reactions [17]. Superox- 3.2. Reaction with Nitric Oxide
ide may also exist in the soils of Mars. The Viking
Landers were unable to detect evidence of life on Superoxide reacts with nitric oxide (NO˙) at an al-
Mars but, instead, found a chemically reactive soil most diffusion-limited rate to form peroxynitrite ani-
capable of decomposing organic molecules. Using on (ONOOˉ), which is frequently called peroxynitrite
EPR spectrometry, Albert S. Yen and coworkers for simplicity. The reaction is as following: O2˙ˉ +
show that superoxide forms directly on Mars-analog NO˙ → ONOOˉ. Peroxynitrite is a potent oxidant
mineral surfaces exposed to ultraviolet radiation un- that causes damage to various biomolecules [24].
der a simulated Martian atmosphere. The superoxide The fast reaction between superoxide and nitric ox-
can explain the reactive nature of the soil and the ap- ide also contributes to the decreased bioavailability

REACTIVE OXYGEN SPECIES ǀ aimsci.com/ros 102 VOLUME 1 ǀ ISSUE 2 ǀ MARCH 2016

 ROS REVIEW ARTICLES
Haber-Weiss reaction, which was first proposed by
Fritz Haber and Joseph Weiss in 1934 [26] (Figure
5). In addition to his contributions to the field of free
radical biology, Fritz Haber received the Nobel Prize
in Chemistry in 1918 for the synthesis of ammonia
from its elements.
In the absence of Fe ions, the above reaction pro-
ceeds slowly. The presence of Fe ions markedly ac-
celerates the reaction to produce hydroxyl radical.
The Fe ion-catalyzed Haber-Weiss reaction can be
written in two sequential sub-reactions: (1) O2˙ˉ +
Fe3+ → O2 + Fe2+, and (2) Fe2+ + H2O2 → Fe3+ +
OH˙ + OHˉ. The second reaction is commonly re-
ferred to as the Fenton reaction, which is also fre-
quently called the Fenton chemistry.
By definition, Fenton chemistry refers to the oxi-
dation of organic substrates by Fe2+ and H2O2. In
1894, Henry J.H. Fenton first observed the oxida-
tion of tartaric acid by H2O2 in the presence of Fe2+
[27], indicating that a potent oxidant(s) is formed by
FIGURE 4. Chemical reactivity of superoxide. As the reaction between H2O2 and Fe2+, which are also
illustrated, superoxide reacts with iron-sulfur clusters known as the Fenton reagent. The Fenton reagent is
in enzymes, leading to enzyme inactivation. It reacts commonly used to oxidize (destroy) organic chemi-
with nitric oxide (NO˙) at a diffuse-limited rate to cals, such as wastes in water. Later in the 1930s,
form peroxynitrite (ONOOˉ), a potent oxidant. Su- Fritz Haber and Joseph Weiss proposed the for-
peroxide is also a reactant of the iron-catalyzed Ha- mation of hydroxyl radical from the Fenton reagent,
ber-Weiss reaction, giving rise to hydroxyl radical, which is known today to be a major ultimate species
the most potent ROS formed in biological systems. responsible for oxidative damage of a wide range of
Spontaneous or superoxide dismutase (SOD)- biomolecules.
catalyzed dismutation of superoxide produces hy- In addition to Fe2+, other transition metal ions,
drogen peroxide, another important ROS. such as cuprous ion (Cu1+), also react with H2O2,
forming hydroxyl radical or its equivalent: M(n-1)+ +
H2O2 → Mn+ + OH˙ + OHˉ, where M denotes the
transition metal ion. For example, Cu1+ + H2O2 →
of nitric oxide under diverse pathophysiological con-
Cu2+ + OH˙ + OHˉ. Such reactions are commonly
ditions. Nitric oxide is generally regarded as a cardi-
referred to as Fenton-type reactions.
ovascular protective molecule, whose deficiency
might contribute, at least partly, to such diseases as
3.4. Membrane Permeability and Protonation
atherosclerosis, hypertension, coronary artery disease,
and erectile dysfunction [25]
Another factor influencing the biological activities of
superoxide is its limited ability to cross biomem-
3.3. Haber-Weiss Reaction and Fenton Reaction
branes due to the negative charge and short half-life
of this free radical species. However, superoxide
In the presence of transition metal ions, such as iron
may cross cell membranes through anion channels,
(Fe) ions, superoxide and hydrogen peroxide togeth-
such as the 4-diisothiocyano-2,2-disulfonic acid stil-
er can give rise to hydroxyl radical (OH˙), the most
bene (DIDS)-sensitive chloride channel [28]. In con-
potent ROS capable of damaging the entire spectrum
trast, perhydroxyl radical, the protonated form of
of biomolecules. The reaction of superoxide and hy-
superoxide, is highly membrane permeable, and is
drogen peroxide in the presence of Fe ions to pro-
also a potent oxidizing species [29]. Perhydroxyl
duce hydroxyl radical is known as iron-catalyzed

REACTIVE OXYGEN SPECIES ǀ aimsci.com/ros 103 VOLUME 1 ǀ ISSUE 2 ǀ MARCH 2016

 ROS REVIEW ARTICLES

FIGURE 6. Spontaneous and SOD-catalyzed dis-

mutation of superoxide. As illustrated, the dismuta-
tion reaction of superoxide is greatly accelerated by
SOD with a reaction rate constant being over three
orders of magnitude greater than that of spontaneous
FIGURE 5. Iron-catalyzed Haber-Weiss reaction. dismutation.
The reaction between superoxide and H2O2 to form
hydroxyl radical is commonly referred to as the Ha-
ber-Weiss reaction. This reaction proceeds slowly 4.1. Phagocyte-Mediated Immunity
and can be dramatically accelerated by the presence
of iron (Fe) ions. The reaction on the right side of the Superoxide produced from phagocytic NADPH oxi-
scheme is also called the Fenton reaction. dase during respiratory burst is an important precur-
sor molecule leading to the formation of more potent
ROS, especially hypochlorous acid (HOCl) and per-
radical has a pKa of 4.8, and as such, may reach high oxynitrite that kill the invading microorganisms.
levels in cellular compartments with high concentra- Dismutation of superoxide generates hydrogen per-
tions of protons, including the mitochondrial inter- oxide, which in turn reacts with chloride ion in the
membrane space. A recent study by Michael A. presence of the phagocyte-derived myeloperoxidase
Trush and coworkers provides direct evidence for the (MPO), forming hypochlorous acid. Likewise, su-
mitochondrial electron transport chain-derived su- peroxide released from the NADPH oxidase reacts
peroxide to exit mitochondria and cross cell mem- with nitric oxide generated by inducible nitric oxide
branes to enter the extracellular space [8]. Exit of synthase (iNOS), resulting in the formation of per-
superoxide from mitochondria and its subsequent oxynitrite. Failure to generate phagocyte-derived su-
crossing of cell membranes to enter extracellular mi- peroxide and related ROS is the major defect in
lieu provide a basis for superoxide to participate in chronic granulomatous disease, causing recurrent in-
diverse intracellular and intercellular processes under fections and granulomatous complications [30].
physiological and pathophysiological conditions.
4.2. Redox Modulation and Signaling

4. SUPEROXIDE IN BIOLOGY AND The role of superoxide in redox regulation in prokar-

MEDICINE yotes is well established. In bacteria, SoxR and SoxS
are activated by superoxide via reversible one-
Superoxide is one of the most extensively investigat- electron oxidation of the [2Fe-2S] cluster and then
ed ROS in biology and medicine. A PubMed search enhance the production of various antioxidant pro-
on January 30, 2016, revealed 82,289 entries con- teins through the soxRS regulon [31, 32].
taining the term ‘superoxide’ in title/abstract. The Superoxide stimulates cell proliferation. Overex-
number of entries for ‘hydrogen peroxide’ and ‘hy- pression of superoxide-producing NADPH oxidase
droxyl radical’ was 42,193 and 10,008, respectively. induces cell transformation [33] though the responsi-
As outlined below, superoxide anion radical plays an ble ROS seems to be hydrogen peroxide derived
important role in both physiological and pathophysi- from the dismutation of superoxide [34]. Superoxide
ological processes. may also relay Ras protein's oncogenic message [35].

REACTIVE OXYGEN SPECIES ǀ aimsci.com/ros 104 VOLUME 1 ǀ ISSUE 2 ǀ MARCH 2016

 ROS REVIEW ARTICLES
4.3. Autoregulation of Superoxide Production The tissue damage may occur as a result of direct ef-
fects of superoxide on molecular targets but more
Superoxide may regulate its own production. On the likely due to the secondary formation of hydrogen
one hand, superoxide activates mitochondrial uncou- peroxide from superoxide dismutation or the for-
pling proteins, leading to decreased formation of su- mation of peroxynitrite from the reaction of superox-
peroxide from the mitochondrial electron transport ide with nitric oxide (Figure 4).
chain [36, 37]. On the other hand, superoxide flux The causal involvement of superoxide in disease
across the endothelial cell plasma membrane occurs pathogenesis is supported by numerous studies
through chloride ion channels and induces intracellu- showing that genetic deletion of SOD causes diverse
lar calcium ion release, which in turn activates mito- disease processes and transgenic overexpression of
chondrial superoxide generation [38]. SOD renders resistance to disease pathogenesis in
animal models [41–43]. The diseases or conditions
4.4. Cell and Tissue Defenses whose pathogenesis is impacted by the alterations of
SOD expression or content involve diverse organs
Superoxide undergoes spontaneous dismutation to and systems, and include aging [44, 45], cancer [46–
form hydrogen peroxide and molecular oxygen with 48], cardiovascular diseases [49, 50], diabetes [42,
‒ ‒
a reaction rate constant of ~5 × 105 M 1s 1 at pH 7.0 51], neurodegeneration [52, 53], immunological dis-
(Figure 6) . The term dismutation reaction refers to a orders [54, 55], pulmonary disorders [56, 57], hepat-
chemical reaction in which the same reactant is both ic disorders [58, 59], gastrointestinal diseases [60,
oxidized and reduced. In superoxide dismutation re- 61], and kidney diseases [51, 62, 63].
action, one molecule of superoxide is oxidized to Although substantial evidence supports a causal
molecular oxygen, and another is reduced to hydro- involvement of superoxide in diverse disease pro-
gen peroxide. The dismutation reaction of superoxide cesses in animal models, the exact role of superoxide
is catalyzed by superoxide dismutase (SOD) with a and SOD in disease pathogenesis in humans remains
‒ ‒ to be established. The only human disease that is,
reaction rate constant of ~1.6 × 109 M 1s 1 which is
over three orders of magnitude greater than that of thus far, known to be affected by mutations in the
spontaneous dismutation (Figure 6). SOD (Cu,ZnSOD) gene is familial amyotrophic lat-
In mammalian species, including humans, there are eral sclerosis [64]. Large-scale, well-designed, ran-
three types of SOD: (1) Cu,ZnSOD, (2) manganese domized controlled trials on using SOD in human
(MnSOD), and (3) extracellular SOD (ECSOD). All disease intervention are currently lacking. Clinical
three forms of SOD catalyze dismutation of superox- research on the role of superoxide in disease process
ide to hydrogen peroxide and molecular oxygen at a is hampered by the limited bioavailability of native
similar reaction rate constant. SOD. In this context, recent development in selective
Recently, NAD(P)H oxidoreductase 1 (NQO1), an SOD biomimetics with favorable pharmacokinetic
enzyme that catalyzes 2-electrone reduction of qui- properties [65] would facility research on the role of
none compounds, has been suggested to scavenge superoxide in human diseases.
biological superoxide possibly by catalyzing its con-
version to hydrogen peroxide [39, 40]. NQO1 ap-
pears to play a role in scavenging superoxide in 5. PERSPECTIVES
tissues, such as vascular endothelium and myocardi-
um, where SOD is relatively deficient and NQO1 is, The past several decades have witnessed substantial
on the other hand, highly expressed [40]. The fate of advances in understanding the chemistry, biology,
cellular superoxide is illustrated in Figure 7. and medicine of superoxide. Its ubiquitous presence
in biological systems, its formation via defined cellu-
4.5. Pathophysiology lar processes, and its unique biochemical properties
make superoxide an important factor involved in di-
Although superoxide possesses physiological func- verse physiological and pathophysiological processes.
tions, dysregulated formation of this oxygen radical Ongoing efforts on both basic and clinical studies of
leads to tissue injury. In fact, formation of superox- superoxide will further increase our understanding of
ide is an important mechanism of oxygen toxicity. this ROS with regard to its significance in biology

REACTIVE OXYGEN SPECIES ǀ aimsci.com/ros 105 VOLUME 1 ǀ ISSUE 2 ǀ MARCH 2016

 ROS REVIEW ARTICLES

FIGURE 7. The cellular fate of mitochondria-derived superoxide. Mitochondrial electron transport chain
(METC) is the major source of cellular superoxide formation. The METC-derived superoxide undergoes
MnSOD-catalyzed dismutation to form H2O2, which is then converted to water by glutathione peroxidase
(GPx) or peroxiredoxin (Prx) in mitochondrial matrix. Superoxide is able to cross the mitochondrial inner
membrane and enter the cytosol, where Cu,ZnSOD catalyzes its dismutation to H2O2, and the H2O2 formed in
the cytosol or escaped from mitochondria is converted to water by enzymes, including GPx, Prx, and catalase
(CAT), in the cytoplasm. CuZnSOD present in the mitochondrial intermembrane space also catalyzes the dis-
mutation of superoxide present in the intermembrane space. ECSOD acts on extracellular superoxide.

and medicine. The coming years will likely witness 3. Fridovich I, Handler P. Xanthine oxidase. V.
the translation of the new knowledge learned from Differential inhibition of the reduction of various
studies with experimental models into the develop- electron acceptors. J Biol Chem 1962; 237:916–21.
ment of mechanistically-based strategies for modu- 4. Knowles PF, Gibson JF, Pick FM, Bray RC.
lating superoxide-mediated biological effects in Electron-spin-resonance evidence for enzymic
humans so as to minimize its deleterious effects reduction of oxygen to a free radical, the
while maintaining the physiological function of this superoxide ion. Biochem J 1969; 111(1):53–8.
oxygen free radical. 5. McCord JM, Fridovich I. Superoxide dismutase: an
enzymic function for erythrocuprein (hemocuprein).
J Biol Chem 1969; 244(22):6049–55.
6. Babior BM, Kipnes RS, Curnutte JT. Biological
REFERENCES defense mechanisms. The production by leukocytes
of superoxide, a potential bactericidal agent. J Clin
1. Pauling L. The discovery of the superoxide radical. Invest 1973; 52(3):741–4.
Trends Biochem Sci 1979; 4(11):N270–N1. doi: doi: 10.1172/JCI107236.
10.1016/0968-0004(79)90203-2. 7. Li YR, Jia Z, Trush MA. Defining ROS in biology
2. Gerschman R, Gilbert DL, Nye SW, Dwyer P, and medicine. Reactive Oxygen Species 2016;
Fenn WO. Oxygen poisoning and x-irradiation: a 1(1):9–21. doi: 10.20455/ros.2016.803.
mechanism in common. Science 1954;
119(3097):623–6.

REACTIVE OXYGEN SPECIES ǀ aimsci.com/ros 106 VOLUME 1 ǀ ISSUE 2 ǀ MARCH 2016

 ROS REVIEW ARTICLES
8. Li Y, Zhu H, Kuppusamy P, Zweier JL, Trush MA. 20. Fridovich I. Superoxide anion radical (O2-.),
Mitochondrial electron transport chain-derived superoxide dismutases, and related matters. J Biol
superoxide exits macrophages: implications for Chem 1997; 272(30):18515–7.
mononuclear cell-mediated pathophysiological 21. Gardner PR, Fridovich I. Superoxide sensitivity of
processes. Reactive Oxygen Species 2016; 1(1):81– the Escherichia coli aconitase. J Biol Chem 1991;
98. doi: 10.20455/ros.2016.815. 266(29):19328–33.
9. Block K, Gorin Y, Abboud HE. Subcellular 22. Gardner PR, Nguyen DD, White CW. Aconitase is
localization of Nox4 and regulation in diabetes. a sensitive and critical target of oxygen poisoning
Proc Natl Acad Sci USA 2009; 106(34):14385–90. in cultured mammalian cells and in rat lungs. Proc
doi: 10.1073/pnas.0906805106. Natl Acad Sci USA 1994; 91(25):12248–52.
10. Kuroda J, Ago T, Matsushima S, Zhai P, Schneider 23. Diaz F, Enriquez JA, Moraes CT. Cells lacking
MD, Sadoshima J. NADPH oxidase 4 (Nox4) is a Rieske iron-sulfur protein have a reactive oxygen
major source of oxidative stress in the failing heart. species-associated decrease in respiratory
Proc Natl Acad Sci USA 2010; 107(35):15565–70. complexes I and IV. Mol Cell Biol 2012;
doi: 10.1073/pnas.1002178107. 32(2):415–29. doi: 10.1128/MCB.06051-11.
11. Dvash E, Har-Tal M, Barak S, Meir O, Rubinstein 24. Radi R. Peroxynitrite, a stealthy biological oxidant.
M. Leukotriene C4 is the major trigger of stress- J Biol Chem 2013; 288(37):26464–72. doi:
induced oxidative DNA damage. Nat Commun 10.1074/jbc.R113.472936.
2015; 6:10112. doi: 10.1038/ncomms10112. 25. Murad F. Shattuck Lecture. Nitric oxide and cyclic
12. McCord JM. Oxygen-derived free radicals in GMP in cell signaling and drug development. N
postischemic tissue injury. N Engl J Med 1985; Engl J Med 2006; 355(19):2003–11. doi:
312(3):159–63. doi: 10.1056/NEJMsa063904.
10.1056/NEJM198501173120305. 26. Haber F, Weiss J. The catalytic decomposition of
13. Lu Y, Cederbaum AI. Alcohol upregulation of hydrogen peroxide by iron salts. Proc R Soc A
CYP2A5: role of reactive oxygen species. Reactive 1934; 147(861):332–51. doi:
Oxygen Species 2016; 1(2):117-30. doi: 10.1098/rspa.1934.0221
10.20455/ros.2016.823. 27. Fenton HJ. Oxidation of tartaric acid in presence of
14. De Henau S, Tilleman L, Vangheel M, Luyckx E, iron. J Chem Soc, Trans 1894; 65, 899-910.
Trashin S, Pauwels M, et al. A redox signalling doi: 10.1039/CT8946500899.
globin is essential for reproduction in 28. Mumbengegwi DR, Li Q, Li C, Bear CE,
Caenorhabditis elegans. Nat Commun 2015; Engelhardt JF. Evidence for a superoxide
6:8782. doi: 10.1038/ncomms9782. permeability pathway in endosomal membranes.
15. Shaked Y, Rose A. Microbiology. Seas of Mol Cell Biol 2008; 28(11):3700–12. doi:
superoxide. Science 2013; 340(6137):1176–7. doi: 10.1128/MCB.02038-07.
10.1126/science.1240195. 29. De Grey AD. HO2˙: the forgotten radical. DNA
16. Diaz JM, Hansel CM, Voelker BM, Mendes CM, Cell Biol 2002; 21(4):251–7. doi:
Andeer PF, Zhang T. Widespread production of 10.1089/104454902753759672.
extracellular superoxide by heterotrophic bacteria. 30. Kuhns DB, Alvord WG, Heller T, Feld JJ, Pike
Science 2013; 340(6137):1223–6. doi: KM, Marciano BE, et al. Residual NADPH oxidase
10.1126/science.1237331. and survival in chronic granulomatous disease. N
17. Georgiou CD, Sun HJ, McKay CP, Grintzalis K, Engl J Med 2010; 363(27):2600–10. doi:
Papapostolou I, Zisimopoulos D, et al. Evidence 10.1056/NEJMoa1007097.
for photochemical production of reactive oxygen 31. Watanabe S, Kita A, Kobayashi K, Miki K. Crystal
species in desert soils. Nat Commun 2015; 6:7100. structure of the [2Fe-2S] oxidative-stress sensor
doi: 10.1038/ncomms8100. SoxR bound to DNA. Proc Natl Acad Sci USA
18. Yen AS, Kim SS, Hecht MH, Frant MS, Murray B. 2008; 105(11):4121–6. doi:
Evidence that the reactivity of the martian soil is 10.1073/pnas.0709188105.
due to superoxide ions. Science 2000; 32. Demple B, Amabile-Cuevas CF. Redox redux: the
289(5486):1909–12. control of oxidative stress responses. Cell 1991;
19. Zhu H. Assays for detecting biological superoxide. 67(5):837–9.
Reactive Oxygen Species 2016; 1(1):65–80. 33. Suh YA, Arnold RS, Lassegue B, Shi J, Xu X,
doi: 10.20455/ros.2015.813. Sorescu D, et al. Cell transformation by the

REACTIVE OXYGEN SPECIES ǀ aimsci.com/ros 107 VOLUME 1 ǀ ISSUE 2 ǀ MARCH 2016

 ROS REVIEW ARTICLES
superoxide-generating oxidase Mox1. Nature 1999; 44. Orr WC, Sohal RS. Extension of life-span by
401(6748):79–82. doi: 10.1038/43459. overexpression of superoxide dismutase and
34. Arnold RS, Shi J, Murad E, Whalen AM, Sun CQ, catalase in Drosophila melanogaster. Science
Polavarapu R, et al. Hydrogen peroxide mediates 1994; 263(5150):1128–30.
the cell growth and transformation caused by the 45. Melov S, Ravenscroft J, Malik S, Gill MS, Walker
mitogenic oxidase Nox1. Proc Natl Acad Sci USA DW, Clayton PE, et al. Extension of life-span with
2001; 98(10):5550–5. doi: superoxide dismutase/catalase mimetics. Science
10.1073/pnas.101505898. 2000; 289(5484):1567–9.
35. Pennisi E. Superoxides relay Ras protein's 46. Kim SH, Kim MO, Gao P, Youm CA, Park HR,
oncogenic message. Science 1997; Lee TS, et al. Overexpression of extracellular
275(5306):1567–8. superoxide dismutase (EC-SOD) in mouse skin
36. Echtay KS, Roussel D, St-Pierre J, Jekabsons MB, plays a protective role in DMBA/TPA-induced
Cadenas S, Stuart JA, et al. Superoxide activates tumor formation. Oncol Res 2005; 15(7–8):333–41.
mitochondrial uncoupling proteins. Nature 2002; 47. Kensler TW, Bush DM, Kozumbo WJ. Inhibition
415(6867):96–9. doi: 10.1038/415096a. of tumor promotion by a biomimetic superoxide
37. Echtay KS, Murphy MP, Smith RA, Talbot DA, dismutase. Science 1983; 221(4605):75–7.
Brand MD. Superoxide activates mitochondrial 48. Van Remmen H, Ikeno Y, Hamilton M, Pahlavani
uncoupling protein 2 from the matrix side: studies M, Wolf N, Thorpe SR, et al. Life-long reduction
using targeted antioxidants. J Biol Chem 2002; in MnSOD activity results in increased DNA
277(49):47129–35. damage and higher incidence of cancer but does
doi: 10.1074/jbc.M208262200. not accelerate aging. Physiol Genomics 2003;
38. Hawkins BJ, Madesh M, Kirkpatrick CJ, Fisher 16(1):29–37. doi:
AB. Superoxide flux in endothelial cells via the 10.1152/physiolgenomics.00122.2003.
chloride channel-3 mediates intracellular signaling. 49. Yen HC, Oberley TD, Vichitbandha S, Ho YS, St
Mol Biol Cell 2007; 18(6):2002–12. doi: Clair DK. The protective role of manganese
10.1091/mbc.E06-09-0830. superoxide dismutase against adriamycin-induced
39. Siegel D, Gustafson DL, Dehn DL, Han JY, acute cardiac toxicity in transgenic mice. J Clin
Boonchoong P, Berliner LJ, et al. Invest 1996; 98(5):1253–60. doi:
NAD(P)H:quinone oxidoreductase 1: role as a 10.1172/JCI118909.
superoxide scavenger. Mol Pharmacol 2004; 50. Lebovitz RM, Zhang H, Vogel H, Cartwright J, Jr.,
65(5):1238–47. doi: 10.1124/mol.65.5.1238. Dionne L, Lu N, et al. Neurodegeneration,
40. Zhu H, Jia Z, Mahaney JE, Ross D, Misra HP, myocardial injury, and perinatal death in
Trush MA, et al. The highly expressed and mitochondrial superoxide dismutase-deficient mice.
inducible endogenous NAD(P)H:quinone Proc Natl Acad Sci USA 1996; 93(18):9782–7.
oxidoreductase 1 in cardiovascular cells acts as a 51. Kubisch HM, Wang J, Luche R, Carlson E, Bray
potential superoxide scavenger. Cardiovasc TM, Epstein CJ, et al. Transgenic copper/zinc
Toxicol 2007; 7(3):202–11. doi: 10.1007/s12012- superoxide dismutase modulates susceptibility to
007-9001-z. type I diabetes. Proc Natl Acad Sci USA 1994;
41. Melov S, Schneider JA, Day BJ, Hinerfeld D, 91(21):9956–9.
Coskun P, Mirra SS, et al. A novel neurological 52. Murakami K, Murata N, Noda Y, Tahara S,
phenotype in mice lacking mitochondrial Kaneko T, Kinoshita N, et al. SOD1 (copper/zinc
manganese superoxide dismutase. Nat Genet 1998; superoxide dismutase) deficiency drives amyloid
18(2):159–63. doi: 10.1038/ng0298-159. beta protein oligomerization and memory loss in
42. Shen X, Zheng S, Metreveli NS, Epstein PN. mouse model of Alzheimer disease. J Biol Chem
Protection of cardiac mitochondria by 2011; 286(52):44557–68. doi:
overexpression of MnSOD reduces diabetic 10.1074/jbc.M111.279208.
cardiomyopathy. Diabetes 2006; 55(3):798–805. 53. Massaad CA, Washington TM, Pautler RG, Klann
43. Li Y, Huang TT, Carlson EJ, Melov S, Ursell PC, E. Overexpression of SOD-2 reduces hippocampal
Olson JL, et al. Dilated cardiomyopathy and superoxide and prevents memory deficits in a
neonatal lethality in mutant mice lacking mouse model of Alzheimer's disease. Proc Natl
manganese superoxide dismutase. Nat Genet 1995; Acad Sci USA 2009; 106(32):13576–81. doi:
11(4):376–81. doi: 10.1038/ng1295-376. 10.1073/pnas.0902714106.

REACTIVE OXYGEN SPECIES ǀ aimsci.com/ros 108 VOLUME 1 ǀ ISSUE 2 ǀ MARCH 2016

 ROS REVIEW ARTICLES
54. Marikovsky M, Ziv V, Nevo N, Harris-Cerruti C, 60. Carroll IM, Andrus JM, Bruno-Barcena JM,
Mahler O. Cu/Zn superoxide dismutase plays Klaenhammer TR, Hassan HM, Threadgill DS.
important role in immune response. J Immunol Anti-inflammatory properties of Lactobacillus
2003; 170(6):2993–3001. gasseri expressing manganese superoxide
55. Iuchi Y, Okada F, Takamiya R, Kibe N, Tsunoda S, dismutase using the interleukin 10-deficient mouse
Nakajima O, et al. Rescue of anaemia and model of colitis. Am J Physiol Gastrointest Liver
autoimmune responses in SOD1-deficient mice by Physiol 2007; 293(4):G729–38. doi:
transgenic expression of human SOD1 in 10.1152/ajpgi.00132.2007.
erythrocytes. Biochem J 2009; 422(2):313–20. doi: 61. Horie Y, Wolf R, Flores SC, McCord JM, Epstein
10.1042/BJ20090176. CJ, Granger DN. Transgenic mice with increased
56. Folz RJ, Abushamaa AM, Suliman HB. copper/zinc-superoxide dismutase activity are
Extracellular superoxide dismutase in the airways resistant to hepatic leukostasis and capillary no-
of transgenic mice reduces inflammation and reflow after gut ischemia/reperfusion. Circ Res
attenuates lung toxicity following hyperoxia. J Clin 1998; 83(7):691–6.
Invest 1999; 103(7):1055–66. doi: 62. DeRubertis FR, Craven PA, Melhem MF, Salah
10.1172/JCI3816. EM. Attenuation of renal injury in db/db mice
57. Yao H, Arunachalam G, Hwang JW, Chung S, overexpressing superoxide dismutase: evidence for
Sundar IK, Kinnula VL, et al. Extracellular reduced superoxide− nitric oxide interaction.
superoxide dismutase protects against pulmonary Diabetes 2004; 53(3):762–8.
emphysema by attenuating oxidative fragmentation 63. Yin M, Wheeler MD, Connor HD, Zhong Z,
of ECM. Proc Natl Acad Sci USA 2010; Bunzendahl H, Dikalova A, et al. Cu/Zn-
107(35):15571–6. doi: 10.1073/pnas.1007625107. superoxide dismutase gene attenuates ischemia-
58. Wheeler MD, Nakagami M, Bradford BU, Uesugi reperfusion injury in the rat kidney. J Am Soc
T, Mason RP, Connor HD, et al. Overexpression of Nephrol 2001; 12(12):2691–700.
manganese superoxide dismutase prevents alcohol- 64. Rosen DR, Siddique T, Patterson D, Figlewicz DA,
induced liver injury in the rat. J Biol Chem 2001; Sapp P, Hentati A, et al. Mutations in Cu/Zn
276(39):36664–72. doi: 10.1074/jbc.M105352200. superoxide dismutase gene are associated with
59. Laukkanen MO, Leppanen P, Turunen P, Tuomisto familial amyotrophic lateral sclerosis. Nature 1993;
T, Naarala J, Yla-Herttuala S. EC-SOD gene 362(6415):59–62. doi: 10.1038/362059a0.
therapy reduces paracetamol-induced liver damage 65. Bird HZ, Hopkins RZ. Nanomaterials for selective
in mice. J Gene Med 2001; 3(4):321–5. doi: superoxide dismutation. Reactive Oxygen Species
10.1002/jgm.194. 2016; 1(1):59–64. doi: 10.20455/ros.2016.811.

REACTIVE OXYGEN SPECIES ǀ aimsci.com/ros 109 VOLUME 1 ǀ ISSUE 2 ǀ MARCH 2016