Arginine/Arginase NO NO NO

Godfrey S. Getz and Catherine A. Reardon

Arterioscler Thromb Vasc Biol. 2006;26:237-239
doi: 10.1161/01.ATV.0000202014.54609.9d
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Editorial
Arginine/Arginase NO NO NO
Godfrey S. Getz, Catherine A. Reardon

rginine is a semi-essential amino acid that plays a

critical role in several metabolic pathways. The best
known of these is as the immediate precursor of urea
in the liver and to a lesser extent in the intestine.1 The enzyme
responsible for the cleavage of arginine to produce urea is
arginase, which is the only enzyme in the urea cycle that
comes in two forms, arginase I, a cytosolic enzyme, and
arginase II, a mitochondrial enzyme. These enzymes exist not
only in the liver and intestine where urea is generated but are
also widely distributed in the body. Ornithine, the other
product of the cleavage of arginine by arginase, is a precursor
of proline and polyamines such as spermine, spermidine, and
putrescine. Arginine is also the precursor of nitric oxide
(NO).

See page 365

In an important article authored by Teupser and colleagues,2 arginase in macrophages has been identified as a
candidate gene influencing atheroresponsiveness. They created and studied two strains of rabbits with genetically
determined high (HAR) and low (LAR) atherosclerosis susceptibility. Subtractive suppression hybridization was used to
screen for genes that were more highly expressed in macrophages from LAR rabbits. Among the 42 clones identified
were four differentially expressed genes with potential relevance to atherogenesis. These were OC2 protein, fibronectin,
ferritin H-chain, and arginase I. Despite their identification in
the screen, the first three genes were not differentially
expressed in macrophages between HAR and LAR animals
when checked by quantitative RT-PCR. In contrast, arginase
I was more highly expressed in LAR than HAR macrophages,
and its expression level was correlated with higher arginase
enzymatic activity. They further identified a length polymorphism of the mRNA for arginase I that was attributable to
the presence (long variant) or absence (short variant) of a
413-bp C repeat in the 3 untranslated region. The short
variant was expressed highly in the LAR rabbit macrophages
and was associated with significantly higher arginase I
mRNA levels and arginase activity. It was also shown that the
long transcript variant was less stable than the short variant.
Homozygosity for the long variant was found only among the
HAR rabbits, whereas homozygosity for the short variant was
primarily found in LAR rabbits. Though arginase promoter
variants were also noted, these were not related to arginase
From the Department of Pathology, University of Chicago, Chicago, Ill.
Correspondence to Godfrey S. Getz, University of Chicago, Department of Pathology MC 1089, 5841 S Maryland Avenue, Chicago, IL
60637. E-mail g-getz@uchicago.edu
(Arterioscler Thromb Vasc Biol. 2006;26:237-239.)
2006 American Heart Association, Inc.
Arterioscler Thromb Vasc Biol. is available at http://www.atvbaha.org
DOI: 10.1161/01.ATV.0000202014.54609.9d

expression levels. Although plausible, the notion that variation in arginase I activity accounts for the difference in
atherosclerosis susceptibility between HAR and LAR rabbits
has yet to be fully proven. This will likely be done by creating
arginase I genetic variants in a uniform genetic background,
which will allow for a more rigorous quantitative assessment
of the role of arginase activity in contributing to
atherosusceptibility.
The plausibility of arginase as a candidate gene for
atherosusceptibility rests on suggestive evidence in the literature. Arginine administration has been shown to improve
vascular reactivity and to be atheroprotective.3 In addition to
being an important amino acid for protein synthesis and as the
terminal intermediate in ureogenesis, arginine is also the
precursor for the formation of NO by nitric oxide synthases
(NOS)4 (see Figure). The generation of NO in the endothelium results in vasorelaxation and is generally regarded as
atheroprotective, reducing adhesion molecule expression.
Vascular responsiveness to NO is reduced in the presence of
such atherosclerosis risk factors as hypercholesterolemia,
hypertension, and diabetes, and this can be reversed by
administration of arginine. Endothelial nitric oxide synthase
(eNOS) is a highly regulated enzymatic activity, located
primarily in the caveolae of the endothelial cell. It is regulated
by shear stress and is more highly expressed in the endothelium underlying laminar flow of blood.5 It is also regulated by
HDL in an SR-BI dependent fashion6 and by estrogen. NO
can interact with superoxide to form the oxidant peroxynitrite
(ONOO). This happens in the endothelium particularly
when tetrahydrobiopterin, the cofactor for eNOS, is limiting,
resulting in the so called uncoupling of eNOS.7
The work of Teupser and colleagues focused on gene
expression in the macrophage because it is a central cell
involved in atherogenesis. Arginase was identified in human
atherosclerotic plaques mainly colocalized with the macrophages around necrotic cores. In macrophages the major NOS
is the inducible nitric oxide synthase (iNOS) which generates
NO and peroxynitrite in the presence of superoxide. Thus in
contrast to eNOS, the formation of NO by iNOS is mainly
proinflammatory and hence proatherogenic. If higher arginase activity reduced the available arginine substrate for NO
production by iNOS and consequent peroxynitrite, this could
be one mechanism for an atheroprotective effect of high
macrophage arginase activity. It is of interest that the antiinflammatory cytokines interleukin (IL)-4, IL-10, and IL-13 are
capable of inducing arginase I synthesis, with IL-4 and IL-10
acting cooperatively.8,9 This could be one mechanism by
which the Th2 T cells and regulatory T cells through the
production of their prototypic cytokines may exert their
atheroprotective action. Interestingly, these cytokines influence macrophage arginase I expression, but not arginase II.10

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238

Arterioscler Thromb Vasc Biol.

February 2006

Arginine can be used for protein synthesis or alternatively it can

be metabolized by NOS pathways to generate NO or cleaved by
arginase to ornithine and urea. Arginase I synthesis is increased
by antiinflammatory cytokines. Increased arginase I activity in
macrophages may be antiatherogenic by reducing NO production leading to reduced generation of peroxynitrite (ONOO).
Reduced arginine levels, although still above the Km for NOS,
decreases NO generation probably by reduced translation of
NOS or reduced competitive displacement of the competitive
inhibitor ADMA (asymmetrical dimethyl arginine). This is known
as the arginine paradox. On the other hand, increased arginase I
activity in endothelial cells could be proatherogenic because of
decreased endothelial cell NO production, and in smooth muscle cells this could result in increased production of collagen
and enhanced cell proliferation attributable to the metabolism of
ornithine into proline and polyamines, respectively.

Endothelial cells and macrophages are not the only cells of

the atherosclerotic plaque involved in the arginaseNO network. The smooth muscle cell also produces arginase I.
Ornithine, one of the products of arginase activity, is the
precursor of proline, glutamate, and polyamines. Proline is a
major substrate for collagen synthesis. The polyamines promote cell cycle progression and therefore cell proliferation.11,12 Both of these processes could influence the nature
and composition of the atherosclerotic plaque.
Variation in arginase type and activity may also be expressed in endothelial cells and smooth muscle cells as well
as in macrophages, though this is yet to be explicitly shown.
Alterations in the expression of arginase I in these vascular
wall cells may have complex effects on atherogenesis. One
might predict that increased arginase in endothelial cells
would not be atheroprotective because of decreased NO
production and decreased vasorelaxation, whereas expression
in smooth muscle cells could influence the cellular matrix
composition of an atherosclerotic plaque as a result of
increased generation of proline and polyamines. On the other
hand, increased expression in macrophages and smooth
muscle cells may lead to decreased production of peroxynitrite after NO production by iNOS. The contribution of
arginase expression by each of these cell types can only be
resolved by experiments with the selective overexpression of
arginase in endothelial cells, macrophages, or smooth muscle

cells. The phenotypes of tissue specific arginase transgenic

mice crossed with a standard murine atherosclerosis model,
eg, apoE-deficient mouse, would be of interest.
The long and short variant arginase transcripts in rabbit
depends on the presence or absence of the C-type repeat in the
3 untranslated region of the transcript, affecting message
stability. The C-type repeat belongs to a family of short
interspersed nucleotide elements or SINES.13,14 This may be
unique to the rabbit and perhaps other lagomorphs because
other species have different types of SINES. For example,
SINES belonging to B1 and B2 subfamilies are found in
mice.15,16 Thus, there is no current basis for anticipating the
variation in arginase I transcripts will be duplicated in the
mouse.
What is particularly important and attractive about the
work on rabbit arginase is that it shows that quantitative
variation in macrophage arginase activity correlates with
different atherosclerosis responses. However, the role of
arginase in competing for arginine substrate otherwise available for NOS is more complex than might be apparent. The
arginine Km for NOS is much lower than the intracellular
concentration of arginine in most cells.17 The Km for arginase
is much higher, though its Vmax is much higher than that of
iNOS. With this in mind, it is not straightforward that
modifying the availability of arginine, either by providing or
removing exogenous arginine or by degrading intracellular
arginine as a result of increased expression of arginase, would
affect iNOS activity. This has been designated the arginine
paradox. That arginine availability may influence NO generation is clearly illustrated by the phenotype of rare genetic
arginine deficiency in humans.18 Two explanations for the
arginine paradox have been offered. Decreased availability of
arginine influences the synthesis of iNOS not by direct
competition for substrate but by interfering with iNOS
mRNA translation. Both arginine and leucine are required for
initiation of protein synthesis and the level of arginine
influences the phosphorylation and activity of eukaryotic
initiation factor.19 Another possible explanation is that arginine competes with asymmetrical dimethyl arginine
(ADMA), an inhibitor of iNOS and eNOS.20,21 ADMA is
generated by the proteolysis of ADMA containing proteins.
In conclusion, the work of Teupser and colleagues highlights the potential importance of arginine in vascular inflammation and atherosclerosis via iNOS, and in vascular response and remodeling. A great deal of work remains to be
done to unravel all the mechanisms at work, especially in
relation to the effect of arginine metabolism in the participant
cells during atherogenesis. Although depletion of arginine by
arginase is here implicated in atherogenesis, it may also
influence other pathologies, eg, immune function and asthma.22 Because quantitative variation in arginase activity
appears to impact on atherosusceptibility, it will be of interest
to ascertain whether mice wild type for arginase or heterozygous for arginase deficiency exhibit differences in
atherosusceptibility.

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