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Vol. 299, Issue 3, 818-824, December 2001
Department of Pharmacology and Molecular Cardiobiology Program, Boyer Center for Molecular Medicine, Yale University School of Medicine, New Haven, Connecticut
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Abstract |
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 Top
 Abstract
 Introduction
Negative Regulatory Proteins
Positive Regulatory Proteins
Summary
References
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Endothelial nitric oxide synthase (eNOS) is important for cardiovascular homeostasis, vessel remodeling, and angiogenesis. Given the impact of endothelium- derived nitric oxide (NO) in vascular biology, much work in the past several years has focused on the control of NO synthesis by regulatory proteins that influence its function. Indeed calcium-activated calmodulin is important for regulation of NOS activity. Herein we discuss why other proteins, in addition to calmodulin, are necessary for eNOS regulation and summarize the biology of negative and positive regulators of eNOS function in vitro, in cells, and in blood vessels.
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Introduction |
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Abstract
Introduction
Negative Regulatory Proteins
Positive Regulatory Proteins
Summary
References
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Endothelium-derived
nitric oxide (NO), the classic relaxing factor discovered by Furchgott
in 1980, is produced by the enzyme endothelial nitric oxide synthase
(eNOS). Early observations by pharmacologists studying
endothelium-dependent relaxations of blood vessels and the release of
relaxing factor by cultured endothelial cells demonstrated that removal
of extracellular calcium from media solutions blocked agonist-induced
release of NO, suggesting that calcium was important for the release of
endothelium-derived NO (Singer and Peach, 1982
; Forstermann et al.,
1991). Additional studies using broken cell systems documented that
calcium removal or antagonism of calmodulin (CaM) with inhibitors
blocked the generation of NO and NOS activity, suggesting that eNOS was
a calcium-calmodulin-requiring enzyme (Busse and Mulsch, 1990;
Forstermann et al., 1991). The requirement for calcium-calmodulin was
proven upon purification of eNOS to homogeneity (Pollock et al., 1991) and rationalized by the presence of a calmodulin binding motif in the
deduced amino acid sequence of the cloned eNOS cDNA.
In the past fours years many laboratories have described proteins other
than CaM that may negatively or positively impact eNOS function.
Insights into the need for additional regulatory proteins important for
NO production from endothelial cells stemmed from observations that
eNOS was an N-myristoyl protein (Pollock et al.,
1992).N-Myristoylation is important for the subcellular targeting of discrete microdomains of cells, and mutations that block
N-myristoylation impede proper subcellular targeting and various aspects of signal transduction. Indeed, expression of a
nonacylated form of eNOS did not affect enzymatic activity in broken
cell lysates but prevented calcium ionophore-stimulated NO release,
arguing that additional mechanisms other than CaM, per se, were
important for the fidelity of signal transduction coupling to eNOS
(Sakoda et al., 1995; Sessa et al., 1995). In addition, the hypothesis
that eNOS had to be localized to proper intracellular membranes to be
near to other regulatory proteins (scaffolds, chaperones, kinases)
provided the rationale for the discovery of additional protein
regulators of eNOS function. Described below are putative regulators
of eNOS function that have been shown to inhibit or enhance eNOS
activity and NO release (Table 1).
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Negative Regulatory Proteins |
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Abstract
Introduction
Negative Regulatory Proteins
Positive Regulatory Proteins
Summary
References
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Caveolin.
Caveolin, being the major coat protein of caveolae,
has several faces that may influence the biology of proteins that
localize to cholesterol-rich plasmalemma caveolae. Indeed caveolin-1 is necessary for the biogenesis of caveolae through an unknown mechanism (Smart et al., 1999). In addition, caveolin-1 can serve as a
cholesterol binding protein and traffic cholesterol from the
endoplasmic reticulum through the Golgi to the plasma membrane.
Finally, caveolin has the capacity to directly interact with other
intracellular proteins such as c-Src and H-Ras through amino acids
82-101, the putative scaffolding domain (Smart et al., 1999). Indeed,
three groups independently demonstrated that eNOS could directly
interact with caveolin-1 or caveolin-3 (Feron et al., 1996;
García-Cardeña et al., 1996; Ju et al., 1997). The
primary binding region of caveolin-1 for eNOS is within amino acids
60-101 and, to a lesser extent, amino acids 135-178 (Garcia-Cardena
et al., 1997; Ju et al., 1997). Furthermore, the caveolin-eNOS
immunocomplex is disrupted in the presence of caveolin scaffolding
peptides (amino acids 82-101) (Michel et al., 1997b).
)
located within amino acids 350-358. The importance of the caveolin
interaction with eNOS has been most reproducibly demonstrated by the
effects of caveolin scaffolding peptides and GST-caveolin on NOS
activity. Incubation of pure eNOS with peptides derived from the
scaffolding domains of caveolin-1 and -3 result in inhibition of eNOS
activity (Garcia-Cardena et al., 1997). In cotransfection experiments,
caveolin over-expression in COS-7 cells resulted in a reduction of eNOS
activity (Michel et al., 1997b), and a reduction in NO release was also
observed (Garcia-Cardena et al., 1997). Furthermore, mutagenesis of the
predicted caveolin binding motif within eNOS blocked the ability of
caveolin to suppress NO release in these latter experiments
(Garcia-Cardena et al., 1997). The reduction of eNOS activity by
caveolin peptides, or over-expressed caveolin, is reversed by exogenous
addition of calmodulin, suggesting a reciprocal regulation of eNOS by
calmodulin, an activator, and caveolin, an inhibitor (Michel et al.,
1997a).
Collectively, these overall results suggest that NO production is
negatively regulated by interactions with caveolin and that for NO
release to occur, the inhibitory clamp by caveolin must be overcome.
CaM has been proposed to be solely responsible for the dissociation of
eNOS from caveolin (Michel et al., 1997a). However, the relationship
between caveolin as an inhibitor of eNOS and CaM as its allosteric
modulator has not been examined in light of new findings demonstrating
a role for other positive and negative regulators of eNOS activation.
Another important issue is that there are no direct data showing more
NO release from cells that do not express caveolins or that disruption
of the eNOS-caveolin complex can lead to increased or prolonged NO release from cells, fundamental experiments if caveolin-1 truly negatively regulates eNOS and NO release.
In Vivo Evidence Supporting the eNOS-Caveolin Interaction.
To
date, caveolin knockout mice are not available; therefore, examining
endothelial function in these mice is not yet feasible. However, recent
work using the caveolin scaffolding domain as a surrogate for caveolin
has demonstrated that eNOS can be regulated in situ. Exposure of
permeabilized cardiac myocytes to the caveolin-3 scaffolding domain
peptide (amino acids 55-74), but not a scrambled version, antagonized
the negative chronotropic actions of carbachol (Feron et al., 1998).
Our group recently used a membrane-permeable form of the caveolin-1
scaffolding domain (amino acids 82-101) by fusing it to a
cell-permeable leader sequence (Bucci et al., 2000a). Exposure of the
peptide to blood vessels resulted in uptake into the endothelium and
adventitia and blockade of ACh-induced relaxations, with no effect on
relaxant responses to sodium nitroprusside or the release of
prostacyclin, showing that in an intact blood vessel, the caveolin
peptide is a potent inhibitor of eNOS. In addition, the peptide also
blocked inflammation in two different models by influencing vascular
permeability, suggesting that peptidomimetics may be useful
therapeutically. With respect to disease mechanisms that may influence
the caveolin/eNOS interaction, there is evidence that in a rat model of
cirrhosis, caveolin-1 is over-expressed, more caveolin-1 interacts with
eNOS, and the basal and stimulated production of NO is depressed (Shah
et al., 1999a), suggesting that this interaction may increase portal
pressures and contribute to the disease state.
Intracellular Domains of G-Protein-Coupled Receptors.
Work by
Venema et al. (1996) has shown that the intracellular domain 4 (ID4) of
the bradykinin 2 (B2) and the angiotensin II R1 receptors can
negatively regulate eNOS activity in vitro (Ju et al., 1998). Indeed,
eNOS coprecipitated with the B2 receptor and in vitro interacted with a
GST fusion of ID4, and synthetic peptides from ID4 inhibited eNOS
activity in a dose-dependent manner in vitro. Mechanistically, the ID4
peptide has been shown to affect NOS catalysis by interference with
flavin to heme electron transfer (Golser et al., 2000). The concept
that a receptor can directly interact with eNOS is extremely novel,
suggesting that signaling, albeit negative signaling to eNOS, can occur
in the absence of a G-protein intermediate. However, direct evidence supporting the physiological relevance of this interaction is presently unavailable.
NOSIP.
NOSIP is the newest protein to interact with eNOS
(Dedio et al., 2001). NOSIP is a 34-kDa protein that was initially
identified as an eNOS binding partner. The interaction between NOSIP
and eNOS has been shown both in vitro and in vivo, and through deletion analysis, NOSIP was shown to bind eNOS between amino acids 366 and 486. Stimulation of cells with calcium ionophore does not change the
association of NOSIP and eNOS; however, a peptide derived from the
scaffolding domain of caveolin (82-101) is able to displace eNOS from
NOSIP. NOSIP does not affect eNOS activity assays in vitro but, when
coexpressed in cells, does reduce ionomycin-stimulated NO release. The
ability of NOSIP to reduce NO release from intact cells is due to the
redistribution of eNOS from the plasma membrane to intracellular
compartments. The specificity of NOSIP to eNOS and the true function of
NOSIP are not known.
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Positive Regulatory Proteins |
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Top
Abstract
Introduction
Negative Regulatory Proteins
Positive Regulatory Proteins
Summary
References
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Calmodulin.
The first protein shown to be involved in eNOS
regulation was calmodulin (CaM). Early studies on neuronal NOS and eNOS
(Bredt and Snyder, 1990; Forstermann et al., 1991), demonstrated that purified NOS utilized CaM as an activator of NO synthesis.
Mechanistically, CaM binding to a canonical CaM binding motif can
displace an adjacent autoinhibitory loop on eNOS and neuronal
NOS, thus facilitating NADPH-dependent electron flux from the
reductase domain of the protein through to the oxygenase domain. The
terminal electron acceptor in the oxygenase domain is heme, which can
bind oxygen for insertion into the NOS substrate,
L-arginine. To date, there are no
papers documenting that CaM can actually be recruited to eNOS in a
stimulus-dependent manner and that the recruitment occurs contemporaneously with NO release. Pharmacological evidence using inhibitors of CaM or calcium-free buffers have indirectly shown the
requirement for CaM. Recent work has shown that eNOS immunoprecipitated from human endothelial cells has immunoreactive CaM bound to it (Russell et al., 2000). Moreover, upon challenge of the cells with
estrogen, the amount of CaM recovered in the eNOS immunocomplex does
not change. This suggests that CaM may serve as a tightly bound
prosthetic group, akin to CaM found in inducible NOS (Cho et al.,
1992), and that regulation of the affinity of CaM interactions with NOS
may occur through subtle changes in free calcium levels.
Heat Shock Protein 90 (hsp90).
The hsp90 family is a group of
highly conserved stress proteins that are expressed in all eukaryotic
cells (Pratt, 1997). Two genes encode hsp90, with the human gene
products hsp90
and hsp90
having 86% sequence homology. The hsp90
is highly abundant in cells, accounting for 1 to 2% of cytosolic
protein, and is localized to the cytoplasm, with a small amount found
in the nucleus and cytoskeleton (Pratt, 1997). The main function of
hsp90 has been its involvement in a multicomponent chaperone system
that is responsible for the proper folding of proteins such as steroid receptors and cell cycle-dependent kinases (Pratt, 1997) However, the
abundance of hsp90 associated with newly synthesized proteins suggests
that this may not be its only function. There is increasing evidence
that hsp90 may be an integral part of signal transduction in all cells.
Indeed, hsp90 orthologs are important for tyrosine kinase signaling in
Drosophila and receptor-G-protein signaling in yeast.
; Garcia-Cardena et al., 1998). Indeed, hsp90 was associated
with eNOS in resting endothelial cells; and treatment of cells with
four distinct stimuli that cause NO release, vascular endothelial
growth factor (VEGF), histamine, fluid shear stress, and estrogen, all
enhanced the interaction between hsp90 and eNOS in a time frame
consistent with NO release (Garcia-Cardena et al., 1998; Russell et
al., 2000). The rapid stimulus-dependent formation of the hsp90-eNOS
hetero-complexes suggests that it occurs simultaneously with other
signaling events such as the mobilization of intracellular calcium
and/or protein phosphorylation.
The mechanism of how hsp90 regulates eNOS function is less clear.
Previously we have shown that hsp90 can directly activate eNOS in vitro
(Garcia-Cardena et al., 1998), and coexpression of eNOS with hsp90 in
COS cells increased NOS activity in broken cell lysates. These results
suggest that hsp90 may act as an allosteric modulator of eNOS by
inducing a conformational change in the enzyme that results in
increased activity or possibly stabilize the "activation" complex.
A recent paper has shown that hsp90 increases CaM affinity for neuronal
NOS (Song et al., 2001). Alternatively, hsp90 may act as a scaffold for
the recruitment of other regulatory molecules including kinases and
phosphatases that may influence eNOS function.
Interactions Between Caveolin, CaM, and hsp90.
As mentioned
previously, CaM has been proposed to be exclusively responsible for the
dissociation of eNOS from caveolin. Recently the relationship between
caveolin as an inhibitor of eNOS and CaM as its allosteric modulator
has been examined in light of hsp90 as an additional regulatory
protein. Labeling of endothelial cells with
[35S]methionine followed by immunoprecipitation
of eNOS resulted in the appearance of several co-associated
radiolabeled proteins interacting substoichiometrically with eNOS
(Gratton et al., 2000). Western blotting of these immunoprecipitated
proteins demonstrated the presence of eNOS, caveolin-1, and hsp90 in
the same complex. Moreover, the addition of exogenous CaM weakly
displaced caveolin from CaM. Reconstitution of the heterotrimeric
complex in vitro showed the eNOS interaction with both hsp90 and
caveolin, but the latter proteins did not interact with each other,
demonstrating that eNOS was the bridge holding the complex together.
Interestingly, the binding of caveolin to eNOS was displaced by the
caveolin scaffolding domain peptide, but not by calcium-activated CaM, demonstrating that CaM cannot physically disrupt the eNOS-caveolin complex in vitro. However, hsp90, per se, did not influence the eNOS/caveolin interaction but facilitated the ability of CaM to displace caveolin from eNOS. These data are consistent with two potential models: 1) perhaps the "recruitment or activation" of hsp90 and CaM to eNOS results in weak physical displacement of eNOS
from caveolin, but the complex remains in caveolae; or 2) hsp90 and
calcium-activated CaM coexist with eNOS bound to caveolin, and a slight
change in eNOS conformation, in the absence of bulk translocation away
from caveolin, allows for efficient stimulus-response coupling.
In Vivo Evidence Supporting the eNOS-hsp90 Interaction.
To
study the relationship between hsp90-mediated signaling and NO
production, a specific inhibitor of hsp90, the ansamycin antibiotic
geldanamycin (GA) was used. GA binds to the unique ATP site of hsp90
and a related protein, GRP94, and influences the conformational
stability of hsp90 binding to its substrates (Pratt, 1997). GA
attenuated histamine and VEGF-stimulated cGMP production in cultured
endothelial cells and blocked ACh-induced vasorelaxation of rat aortic
rings (Garcia-Cardena et al., 1998), middle cerebral artery (Khurana et
al., 2000), and flow-induced dilation (Viswanathan et al., 1999),
indicating that hsp90 signaling was crucial for NO release and
endothelial function. Further support for the relevance of hsp90/eNOS
interactions in vivo was demonstrated in a model of portal vein
ligation in rats (Shah et al., 1999b) and in a model of inflammation
(Bucci et al., 2000b). In the former study, the physical interaction of
hsp90 with eNOS isolated from the mesenteric microcirculation was
documented, and GA attenuated ACh-dependent vasodilatation to the same
extent as conventional NOS inhibitors. In portal hypertensive rats,
eNOS protein levels are not changed compared with control rats, but NOS
activity is markedly enhanced in the mesenteric tissue of hypertensive
rats. The enhanced activity correlated with hyporesponsiveness to the vasoconstrictor methoxamine, and GA potentiated the methoxamine-induced vasoconstriction after portal vein ligation, partially reversing the
hyporeactivity to this agent, indicating that hsp90 can act as a
signaling component leading to NO-dependent responses in the mesenteric
microcirculation. In the latter study, GA inhibited inflammation in a
dose-dependent manner, an effect as potent as a steroid. Because GA
blocks NO release and NOS inhibitors reduce edema formation, it is
possible that drugs that specifically inhibit hsp90 will be good
anti-inflammatory drugs.
Dynamin-2.
Dynamin-2 belongs to the family of large GTPases
and is believed to be involved in vesicle formation, receptor-mediated
endocytosis, caveolae internalization and vesicle trafficking in and
out of the Golgi. Dynamin-2 has been shown by confocal microscopy to colocalize with eNOS in the Golgi membranes of endothelial cells and to
bind eNOS directly, both in vivo and in vitro (Cao et al., 2000).
Previous work by others has documented dynamin-2 in the plasma membrane
of endothelial cells, suggesting that, like eNOS, dynamin-2 exists in
the plasma membrane and Golgi. Based on extensive in vitro studies,
dynamin-2 has an affinity for eNOS in the nanomolar range. In cells,
the association between eNOS and dynamin-2 is increased by calcium
ionophore, and in in vitro activity assays, dynamin has been shown to
directly augment NOS activity. The physiological role of this
interaction has yet to be explored.
Signaling Kinases: Akt, RAF, and Erk.
Immunoprecipitation of
eNOS from endothelial cells also results in the coprecipitation of
kinases and related proteins including Akt, Erk, and RAF (Michell et
al., 1999; Bernier et al., 2000). The interaction of these proteins
with eNOS distinguishes them from other protein/protein interactions in
that it is the effect of the kinase (phosphorylation) rather than the
presence of the protein that influences eNOS activity. The relevance of
Akt binding is clear based on in vitro and in vivo evidence
describing the phosphorylation site (see below), whereas the
phosphorylation by Erk has not been characterized. However, the
association of these proteins to eNOS following agonist activation is
indicative of a dynamic multiprotein signaling complex influencing eNOS
function (i.e., NOS-osome).
Protein Phosphorylation.
Another post-translational
modification that can potentially regulate eNOS activity is through
protein phosphorylation (Fig. 1; Table
2). eNOS is primarily phosphorylated on
serine residues and to a lesser extent on tyrosine and threonine
residues (Michel et al., 1993; Corson et al., 1996;
García-Cardeña et al., 1996). The ability of
pharmacological inhibitors of phosphatidylinositol 3-kinase (PI-3K;
i.e., wortmannin and LY298004) to reduce insulin- and VEGF-stimulated
NO release provided the first evidence that a downstream effector of
PI-3K could modulate eNOS activity (Zeng and Quon, 1996;
Papapetropoulos et al., 1997). The protein kinase Akt is activated by
the 3-phosphorylated inositol lipids generated by PI-3K, and is known
to phosphorylate a limited number of cellular substrates according to a
distinct substrate motif that is found in eNOS (RXRXXXS/T). Akt can
directly phosphorylate recombinant eNOS or eNOS in situ, at serine 1177 (human)/1179 (bovine) (Dimmeler et al., 1999; Fulton et al., 1999;
Gallis et al., 1999; Michell et al., 1999). Due to the promiscuous
nature of kinase cascades, it is much more difficult to demonstrate
that eNOS is a direct Akt substrate in vivo; however, several lines of
evidence support this idea. Cotransfection of wild-type Akt and eNOS in
COS cells increases eNOS phosphorylation in a wortmannin-sensitive
manner and is inhibited by mutation of serine 1177/1179 to alanine
(Dimmeler et al., 1999; Fulton et al., 1999). Stimulation of
endothelial cells with VEGF and shear stress phosphorylates and
activates Akt, and within a similar time frame, eNOS is phosphorylated
on serine 1179 in a PI-3K-dependent manner (Dimmeler et al., 1999; Fulton et al., 1999; Gallis et al., 1999; Michell et al., 1999). Akt-phosphorylated eNOS is 15- to 20-fold more active than
unphosphorylated eNOS (Gallis et al., 1999) or more active at lower
levels of calcium or calmodulin (Fulton et al., 1999; Michell et al.,
1999). Mutation of serine 1177 to aspartate (S1177D), which mimics the
negative charge afforded by phosphorylation, results in an enzyme that is constitutively active at low levels (10 nM) of calcium (Dimmeler et
al., 1999), whereas mutation of serine 1177/1179 to an alanine prevents
Akt-dependent NO release (Fulton et al., 1999). Adenoviral-mediated gene transfer of constitutively active Akt to endothelial cells markedly increases basal NO release. Activated Akt potentiated and the
activation-deficient Akt inhibited VEGF-stimulated NO release by
approximately 50 to 70%, similar to levels after wortmannin treatment
(Papapetropoulos et al., 1997; Fulton et al., 1999). As stated
previously, the subcellular localization of eNOS is crucial for
agonist-induced NO release; therefore, we tested whether Akt-dependent
activation of eNOS was influenced by its distribution (Fulton et al.,
1999). Transfection of cells with a mutant form of eNOS that cannot be
acylated prevents the ability of Akt to stimulate NO release,
suggesting that eNOS must be membrane-associated for this interaction
to occur, and may explain in part the reduction in agonist-induced NO
release from cells where eNOS is mislocalized (Sessa et al., 1995; Liu
et al., 1996).
|
). Other kinases that phosphorylate serine 1177/1179 are the
AMP-activated kinase, which phosphorylates serine 1177 in addition to
threonine 495 (Chen et al., 1999), protein kinase A, and protein kinase
G (Butt et al., 2000). Inhibition of calcium-calmodulin kinases with
KN-62 does not affect serine 1179 phosphorylation and activation of
cyclic GMP-dependent kinases with NO or cGMP did not induce
phosphorylation of this site (Gallis et al., 1999). It is likely that
other kinases capable of phosphorylating serine 1177/1179 remain to be identified.
Serine 1177/1179 is not the only eNOS phosphorylation site. For
example, shear stress induces the phosphorylation of another site on
eNOS, serine 116 (Gallis et al., 1999). The region surrounding this
site does not conform to the accepted Akt motif, thus the kinase and
the functional significance are not known. eNOS is also phosphorylated
at threonine 495 (497) by AMP-activated kinase and by protein kinase C. Phosphorylation of eNOS at this site reduces eNOS catalytic activity
(Chen et al., 1999). Tyrosine phosphorylation of eNOS results in an
enzyme with less activity; however, the kinase responsible and the
site(s) of phosphorylation are unknown (García-Cardeña et
al., 1996).
Given the demonstrated importance of serine 1177/1179, there are still
unresolved issues: in particular, the mechanism of why phosphorylation
of serine 1179 increases eNOS activity and sensitivity to
calcium/calmodulin in intact cells. It has been speculated that this
may be due to a conformational change and displacement of an
autoinhibitory carboxy tail. Comparison of recombinant wild-type eNOS
to an S1179D mutant to mimic the activated state shows that S1179D eNOS
is a much more efficient NOS due to enhanced electron flux from the
reductase to the oxygenase domain. In addition, the dissociation of CaM
is slower than that of wild-type enzyme. The idea that phosphorylation
on the carboxy tail removes autoinhibition is supported by the
increased activity of truncation mutants of eNOS, terminated at serine
1177 (Chen et al., 1999). It is also possible that phosphorylation
influences the relationship between the carboxy tail and the
calmodulin-autoinhibitory loop (Fulton et al., 1999). It is unlikely
that phosphorylation-induced changes in eNOS activity can be entirely
attributed to changes in eNOS-associated proteins because
recombinant-phosphorylated eNOS and eNOS S1179D are more active than
unphosphorylated eNOS (McCabe et al., 2000).
In Vivo Evidence Supporting a Functional Role for eNOS
Phosphorylation.
A physiological role of endogenous Akt in
vasomotor function has been identified in blood vessels. Akt activity
in the endothelium of blood vessels was modified by adenoviruses
encoding constitutively active Akt, myr-Akt, and a dominant negative
Akt, Akt-AA (Luo et al., 2000). In femoral arteries infected with the
myr-Akt virus, the resting diameter was significantly larger than
control arteries, and the increase in diameter was reversed by
N
-nitro-L-arginine
methyl ester, an inhibitor of nitric oxide synthesis. Also,
myr-Akt-infected blood vessels displayed an increase in baseline blood
flow. Although dominant negative Akt did not significantly affect
resting diameter or blood flow, it did reduce the ability of ACh, but
not nitroglycerin, to increase both diameter and blood flow. These data
strongly indicate that the Akt-eNOS axis is important for blood flow
control in conduit vessels.
), the
improvement in endothelial function may not entirely be attributed to
changes in gene expression (Kaesemeyer et al., 1999). Recently, it has been shown that simvastatin can activate endothelial Akt, leading to an
increase in eNOS phosphorylation and NO release (Kureishi et al.,
2000). Simvastatin treatment resulted in an increase in endothelial
cell survival and angiogenesis in normocholesterolemic animals.
Collectively, these results suggest that endogenous Akt can participate
in the release of NO from the endothelium in response to agonist
stimulation and that increasing Akt activity, either using adenoviruses
or simvastatin, can stimulate NO release.
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Summary |
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Top
Abstract
Introduction
Negative Regulatory Proteins
Positive Regulatory Proteins
Summary
References
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Given the fundamental importance of endothelial-derived NO in cardiovascular homeostasis and physiology, elucidation of the enzymatic control mechanisms by the aforementioned protein regulators will increase our understanding of how NO release is controlled in vivo. In addition, perhaps novel insights into the mechanisms of endothelial dysfunction, a manifestation of many cardiovascular diseases, may be attributable to impairments in upstream protein regulators of eNOS function.
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Note Added in Proof. |
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Recently, two papers have demonstrated
enhanced endothelium-dependent responses in mice lacking the gene for
caveolin-1 consistent with the concept that caveolin-1 negatively
regulates eNOS function (Drab et al., 2001; Razani et al., 2001).
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Footnotes |
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Accepted for publication May 25, 2001.
Received for publication February 2, 2001.
This work is supported by grants from the National Institutes of Health (RO1 HL57665, HL61371, and HL64793 to W.C.S.; T32HL10183 to D.F.) and a Grant-in-Aid from the American Heart Association (National Grant to W.C.S.). J.P.G. is the recipient of a fellowship from the Canadian Institutes of Health Research. W.C.S. is an Established Investigator of the American Heart Association.
Address correspondence to: Dr. William C. Sessa, Yale University School of Medicine, Boyer Center for Molecular Medicine, Rm 436 D, New Haven, CT 06536. E-mail: william.sessa{at}yale.edu
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Abbreviations |
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NO, nitric oxide; eNOS, endothelial NO synthase; ACh, acetylcholine; B2, bradykinin 2; CaM, calmodulin; Erk, extracellular signal-related kinase; GA, geldanamycin; GST, glutathione S-transferase; hsp90, heat shock protein 90; ID4, intracellular domain 4; IGF, insulin-like growth factor; NOSIP, nitric oxide synthase interacting protein; PI-3K, phosphatidylinositol 3-kinase; VEGF, vascular endothelial growth factor.
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References |
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Top
Abstract
Introduction
Negative Regulatory Proteins
Positive Regulatory Proteins
Summary
References
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 R. Martinelli, M. Gegg, R. Longbottom, P. Adamson, P. Turowski, and J. Greenwood ICAM-1-mediated Endothelial Nitric Oxide Synthase Activation via Calcium and AMP-activated Protein Kinase Is Required for Transendothelial Lymphocyte Migration Mol. Biol. Cell, February 1, 2009; 20(3): 995 - 1005. [Abstract] [Full Text] [PDF]
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 C. Chabot, K. Spring, J.-P. Gratton, M. Elchebly, and I. Royal New Role for the Protein Tyrosine Phosphatase DEP-1 in Akt Activation and Endothelial Cell Survival Mol. Cell. Biol., January 1, 2009; 29(1): 241 - 253. [Abstract] [Full Text] [PDF]
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 Md. S. Bhuiyan, N. Shioda, M. Shibuya, Y. Iwabuchi, and K. Fukunaga Activation of Endothelial Nitric Oxide Synthase by a Vanadium Compound Ameliorates Pressure Overload-Induced Cardiac Injury in Ovariectomized Rats Hypertension, January 1, 2009; 53(1): 57 - 63. [Abstract] [Full Text] [PDF]
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W.-Z. Ying, K. Aaron, and P. W. Sanders Dietary Salt Activates an Endothelial Proline-Rich Tyrosine Kinase 2/c-Src/Phosphatidylinositol 3-Kinase Complex to Promote Endothelial Nitric Oxide Synthase Phosphorylation Hypertension, December 1, 2008; 52(6): 1134 - 1141. [Abstract] [Full Text] [PDF]
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 M. J. Shackcloth, A. R. Conant, J. Thekkudan, S. Ghotkar, A. W.M. Simpson, and W. C. Dihmis Attenuation of receptor-dependent and -independent vasoconstriction in the human radial artery Eur. J. Cardiothorac. Surg., October 1, 2008; 34(4): 839 - 844. [Abstract] [Full Text] [PDF]
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Z. Hu, J. Chen, Q. Wei, and Y. Xia Bidirectional Actions of Hydrogen Peroxide on Endothelial Nitric-oxide Synthase Phosphorylation and Function: CO-COMMITMENT AND INTERPLAY OF Akt AND AMPK J. Biol. Chem., September 12, 2008; 283(37): 25256 - 25263. [Abstract] [Full Text] [PDF]
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 M Hennenberg, J Trebicka, T Sauerbruch, and J Heller Mechanisms of extrahepatic vasodilation in portal hypertension Gut, September 1, 2008; 57(9): 1300 - 1314. [Abstract] [Full Text] [PDF]
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R. P. Ilagan, M. Tiso, D. W. Konas, C. Hemann, D. Durra, R. Hille, and D. J. Stuehr Differences in a Conformational Equilibrium Distinguish Catalysis by the Endothelial and Neuronal Nitric-oxide Synthase Flavoproteins J. Biol. Chem., July 11, 2008; 283(28): 19603 - 19615. [Abstract] [Full Text] [PDF]
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 T. Inoue, Y. Suzuki, T. Yoshimaru, and C. Ra Nitric oxide protects mast cells from activation-induced cell death: the role of the phosphatidylinositol-3 kinase-Akt-endothelial nitric oxide synthase pathway J. Leukoc. Biol., May 1, 2008; 83(5): 1218 - 1229. [Abstract] [Full Text] [PDF]
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R. Lopez-Sepulveda, R. Jimenez, M. Romero, M. J. Zarzuelo, M. Sanchez, M. Gomez-Guzman, F. Vargas, F. O'Valle, A. Zarzuelo, F. Perez-Vizcaino, et al. Wine Polyphenols Improve Endothelial Function in Large Vessels of Female Spontaneously Hypertensive Rats Hypertension, April 1, 2008; 51(4): 1088 - 1095. [Abstract] [Full Text] [PDF]
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 E. Mata-Greenwood and D.-B. Chen Racial Differences in Nitric Oxide--Dependent Vasorelaxation Reproductive Sciences, January 1, 2008; 15(1): 9 - 25. [Abstract] [PDF]
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 D. Tang, J. Lu, J. P. Walterscheid, H.-H. Chen, D. A. Engler, T. Sawamura, P.-Y. Chang, H. J. Safi, C.-Y. Yang, and C.-H. Chen Electronegative LDL circulating in smokers impairs endothelial progenitor cell differentiation by inhibiting Akt phosphorylation via LOX-1 J. Lipid Res., January 1, 2008; 49(1): 33 - 47. [Abstract] [Full Text] [PDF]
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H. Cai, D. Liu, and J. G.N. Garcia CaM Kinase II-dependent pathophysiological signalling in endothelial cells Cardiovasc Res, January 1, 2008; 77(1): 30 - 34. [Abstract] [Full Text] [PDF]
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Y. Zhang, V. Brovkovych, S. Brovkovych, F. Tan, B.-S. Lee, T. Sharma, and R. A. Skidgel Dynamic Receptor-dependent Activation of Inducible Nitric-oxide Synthase by ERK-mediated Phosphorylation of Ser745 J. Biol. Chem., November 2, 2007; 282(44): 32453 - 32461. [Abstract] [Full Text] [PDF]
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 C. Csortos, I. Kolosova, and A. D. Verin Regulation of vascular endothelial cell barrier function and cytoskeleton structure by protein phosphatases of the PPP family Am J Physiol Lung Cell Mol Physiol, October 1, 2007; 293(4): L843 - L854. [Abstract] [Full Text] [PDF]
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D. M. Dudzinski and T. Michel Life history of eNOS: Partners and pathways Cardiovasc Res, July 15, 2007; 75(2): 247 - 260. [Abstract] [Full Text] [PDF]
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 G. Li, E. J. Barrett, M. O. Barrett, W. Cao, and Z. Liu Tumor Necrosis Factor-{alpha} Induces Insulin Resistance in Endothelial Cells via a p38 Mitogen-Activated Protein Kinase-Dependent Pathway Endocrinology, July 1, 2007; 148(7): 3356 - 3363. [Abstract] [Full Text] [PDF]
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 C. Long, L. G. Cook, G.-Y. Wu, and B. M. Mitchell Removal of Fkbp12/12.6 From Endothelial Ryanodine Receptors Leads to an Intracellular Calcium Leak and Endothelial Dysfunction Arterioscler. Thromb. Vasc. Biol., July 1, 2007; 27(7): 1580 - 1586. [Abstract] [Full Text] [PDF]
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J.-Z. Sheng and A. P. Braun Small- and intermediate-conductance Ca2+-activated K+ channels directly control agonist-evoked nitric oxide synthesis in human vascular endothelial cells Am J Physiol Cell Physiol, July 1, 2007; 293(1): C458 - C467. [Abstract] [Full Text] [PDF]
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B. Chanrion, C. Mannoury la Cour, F. Bertaso, M. Lerner-Natoli, M. Freissmuth, M. J. Millan, J. Bockaert, and P. Marin From the Cover: Physical interaction between the serotonin transporter and neuronal nitric oxide synthase underlies reciprocal modulation of their activity PNAS, May 8, 2007; 104(19): 8119 - 8124. [Abstract] [Full Text] [PDF]
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J. Garthwaite Neuronal nitric oxide synthase and the serotonin transporter get harmonious PNAS, May 8, 2007; 104(19): 7739 - 7740. [Full Text] [PDF]
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E. Nisoli, E. Clementi, M. O. Carruba, and S. Moncada Defective Mitochondrial Biogenesis: A Hallmark of the High Cardiovascular Risk in the Metabolic Syndrome? Circ. Res., March 30, 2007; 100(6): 795 - 806. [Abstract] [Full Text] [PDF]
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E. D. Motley, K. Eguchi, M. M. Patterson, P. D. Palmer, H. Suzuki, and S. Eguchi Mechanism of Endothelial Nitric Oxide Synthase Phosphorylation and Activation by Thrombin Hypertension, March 1, 2007; 49(3): 577 - 583. [Abstract] [Full Text] [PDF]
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R. M. Touyz Regulation of Endothelial Nitric Oxide Synthase by Thrombin Hypertension, March 1, 2007; 49(3): 429 - 431. [Full Text] [PDF]
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C. Long, L. G. Cook, S. L. Hamilton, G.-Y. Wu, and B. M. Mitchell FK506 Binding Protein 12/12.6 Depletion Increases Endothelial Nitric Oxide Synthase Threonine 495 Phosphorylation and Blood Pressure Hypertension, March 1, 2007; 49(3): 569 - 576. [Abstract] [Full Text] [PDF]
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 D. D. Roberts, J. S. Isenberg, L. A. Ridnour, and D. A. Wink Nitric Oxide and Its Gatekeeper Thrombospondin-1 in Tumor Angiogenesis Clin. Cancer Res., February 1, 2007; 13(3): 795 - 798. [Abstract] [Full Text] [PDF]
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W. O. Sampaio, R. A. Souza dos Santos, R. Faria-Silva, L. T. da Mata Machado, E. L. Schiffrin, and R. M. Touyz Angiotensin-(1-7) Through Receptor Mas Mediates Endothelial Nitric Oxide Synthase Activation via Akt-Dependent Pathways Hypertension, January 1, 2007; 49(1): 185 - 192. [Abstract] [Full Text] [PDF]
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H. Suzuki, K. Eguchi, H. Ohtsu, S. Higuchi, S. Dhobale, G. D. Frank, E. D. Motley, and S. Eguchi Activation of Endothelial Nitric Oxide Synthase by the Angiotensin II Type 1 Receptor Endocrinology, December 1, 2006; 147(12): 5914 - 5920. [Abstract] [Full Text] [PDF]
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H. Sano, K. Hosokawa, H. Kidoya, and N. Takakura Negative Regulation of VEGF-Induced Vascular Leakage by Blockade of Angiotensin II Type 1 Receptor Arterioscler. Thromb. Vasc. Biol., December 1, 2006; 26(12): 2673 - 2680. [Abstract] [Full Text] [PDF]
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C. D. Searles Transcriptional and posttranscriptional regulation of endothelial nitric oxide synthase expression Am J Physiol Cell Physiol, November 1, 2006; 291(5): C803 - C816. [Abstract] [Full Text] [PDF]
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K. Yayama, H. Hiyoshi, D. Imazu, and H. Okamoto Angiotensin II Stimulates Endothelial NO Synthase Phosphorylation in Thoracic Aorta of Mice With Abdominal Aortic Banding Via Type 2 Receptor Hypertension, November 1, 2006; 48(5): 958 - 964. [Abstract] [Full Text] [PDF]
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R. M. Carey and J. Park Role of Angiotensin Type 2 Receptors in Vasodilation of Resistance and Capacitance Vessels Hypertension, November 1, 2006; 48(5): 824 - 825. [Full Text] [PDF]
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M. Saura, C. Zaragoza, C. Bao, B. Herranz, M. Rodriguez-Puyol, and C. J. Lowenstein Stat3 Mediates Interelukin-6 Inhibition of Human Endothelial Nitric-oxide Synthase Expression J. Biol. Chem., October 6, 2006; 281(40): 30057 - 30062. [Abstract] [Full Text] [PDF]
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Y. Chen, M. Medhora, J. R. Falck, K. A. Pritchard Jr, and E. R. Jacobs Mechanisms of activation of eNOS by 20-HETE and VEGF in bovine pulmonary artery endothelial cells Am J Physiol Lung Cell Mol Physiol, September 1, 2006; 291(3): L378 - L385. [Abstract] [Full Text] [PDF]
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B. Roy and J. Garthwaite Nitric oxide activation of guanylyl cyclase in cells revisited PNAS, August 8, 2006; 103(32): 12185 - 12190. [Abstract] [Full Text] [PDF]
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Q. Wei and Y. Xia Proteasome Inhibition Down-regulates Endothelial Nitric-oxide Synthase Phosphorylation and Function J. Biol. Chem., August 4, 2006; 281(31): 21652 - 21659. [Abstract] [Full Text] [PDF]
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 C. Fernandez-Hernando, M. Fukata, P. N. Bernatchez, Y. Fukata, M. I. Lin, D. S. Bredt, and W. C. Sessa Identification of Golgi-localized acyl transferases that palmitoylate and regulate endothelial nitric oxide synthase J. Cell Biol., July 31, 2006; 174(3): 369 - 377. [Abstract] [Full Text] [PDF]
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G. Garthwaite, K. Bartus, D. Malcolm, D. Goodwin, M. Kollb-Sielecka, C. Dooldeniya, and J. Garthwaite Signaling from blood vessels to CNS axons through nitric oxide. J. Neurosci., July 19, 2006; 26(29): 7730 - 7740. [Abstract] [Full Text] [PDF]
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C. E. Wood and D. Giroux Expression of Nitric Oxide Synthase Isoforms in the Ovine Fetal Brain: Alteration by Hormonal and Hemodynamic Stimuli Reproductive Sciences, July 1, 2006; 13(5): 329 - 337. [Abstract] [PDF]
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X.-P. Zhang and T. H. Hintze cAMP signal transduction induces eNOS activation by promoting PKB phosphorylation Am J Physiol Heart Circ Physiol, June 1, 2006; 290(6): H2376 - H2384. [Abstract] [Full Text] [PDF]
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Q. Zhang, J. E. Church, D. Jagnandan, J. D. Catravas, W. C. Sessa, and D. Fulton Functional Relevance of Golgi- and Plasma Membrane-Localized Endothelial NO Synthase in Reconstituted Endothelial Cells Arterioscler. Thromb. Vasc. Biol., May 1, 2006; 26(5): 1015 - 1021. [Abstract] [Full Text] [PDF]
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 P. Kleinbongard, R. Schulz, T. Rassaf, T. Lauer, A. Dejam, T. Jax, I. Kumara, P. Gharini, S. Kabanova, B. Ozuyaman, et al. Red blood cells express a functional endothelial nitric oxide synthase Blood, April 1, 2006; 107(7): 2943 - 2951. [Abstract] [Full Text] [PDF]
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R. d. di Villa Bianca, R. Sorrentino, R. Sorrentino, C. Imbimbo, A. Palmieri, F. Fusco, M. Maggi, R. De Palma, G. Cirino, and V. Mirone Sphingosine 1-Phosphate Induces Endothelial Nitric-Oxide Synthase Activation through Phosphorylation in Human Corpus Cavernosum J. Pharmacol. Exp. Ther., February 1, 2006; 316(2): 703 - 708. [Abstract] [Full Text] [PDF]
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P. Oishi, A. Grobe, E. Benavidez, B. Ovadia, C. Harmon, G. A. Ross, K. Hendricks-Munoz, J. Xu, S. M. Black, and J. R. Fineman Inhaled nitric oxide induced NOS inhibition and rebound pulmonary hypertension: a role for superoxide and peroxynitrite in the intact lamb Am J Physiol Lung Cell Mol Physiol, February 1, 2006; 290(2): L359 - L366. [Abstract] [Full Text] [PDF]
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J. E. Church and D. Fulton Differences in eNOS Activity Because of Subcellular Localization Are Dictated by Phosphorylation State Rather than the Local Calcium Environment J. Biol. Chem., January 20, 2006; 281(3): 1477 - 1488. [Abstract] [Full Text] [PDF]
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M. Herrera, G. Silva, and J. L. Garvin A High-Salt Diet Dissociates NO Synthase-3 Expression and NO Production by the Thick Ascending Limb Hypertension, January 1, 2006; 47(1): 95 - 101. [Abstract] [Full Text] [PDF]
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X. Jiang, F. Yang, H. Tan, D. Liao, R. M. Bryan Jr, J. K. Randhawa, R. E. Rumbaut, W. Durante, A. I. Schafer, X. Yang, et al. Hyperhomocystinemia Impairs Endothelial Function and eNOS Activity via PKC Activation Arterioscler. Thromb. Vasc. Biol., December 1, 2005; 25(12): 2515 - 2521. [Abstract] [Full Text] [PDF]
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C. Cheng, R. van Haperen, M. de Waard, L. C. A. van Damme, D. Tempel, L. Hanemaaijer, G. W. A. van Cappellen, J. Bos, C. J. Slager, D. J. Duncker, et al. Shear stress affects the intracellular distribution of eNOS: direct demonstration by a novel in vivo technique Blood, December 1, 2005; 106(12): 3691 - 3698. [Abstract] [Full Text] [PDF]
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D. Jagnandan, W. C. Sessa, and D. Fulton Intracellular location regulates calcium-calmodulin-dependent activation of organelle-restricted eNOS Am J Physiol Cell Physiol, October 1, 2005; 289(4): C1024 - C1033. [Abstract] [Full Text] [PDF]
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J. S. Isenberg, L. A. Ridnour, E. M. Perruccio, M. G. Espey, D. A. Wink, and D. D. Roberts Thrombospondin-1 inhibits endothelial cell responses to nitric oxide in a cGMP-dependent manner PNAS, September 13, 2005; 102(37): 13141 - 13146. [Abstract] [Full Text] [PDF]
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A. Martinez-Ruiz, L. Villanueva, C. G. de Orduna, D. Lopez-Ferrer, M. A. Higueras, C. Tarin, I. Rodriguez-Crespo, J. Vazquez, and S. Lamas S-nitrosylation of Hsp90 promotes the inhibition of its ATPase and endothelial nitric oxide synthase regulatory activities PNAS, June 14, 2005; 102(24): 8525 - 8530. [Abstract] [Full Text] [PDF]
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P. L. Huang Unraveling the Links Between Diabetes, Obesity, and Cardiovascular Disease Circ. Res., June 10, 2005; 96(11): 1129 - 1131. [Full Text] [PDF]
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 V. Mollace, C. Muscoli, E. Masini, S. Cuzzocrea, and D. Salvemini Modulation of Prostaglandin Biosynthesis by Nitric Oxide and Nitric Oxide Donors Pharmacol. Rev., June 1, 2005; 57(2): 217 - 252. [Abstract] [Full Text] [PDF]
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 J. Zheng, I. M. Bird, D.-B. Chen, and R. R. Magness Angiotensin II regulation of ovine fetoplacental artery endothelial functions: interactions with nitric oxide J. Physiol., May 15, 2005; 565(1): 59 - 69. [Abstract] [Full Text] [PDF]
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Q. Wei and Y. Xia Roles of 3-Phosphoinositide-dependent Kinase 1 in the Regulation of Endothelial Nitric-oxide Synthase Phosphorylation and Function by Heat Shock Protein 90 J. Biol. Chem., May 6, 2005; 280(18): 18081 - 18086. [Abstract] [Full Text] [PDF]
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H. Hiyoshi, K. Yayama, M. Takano, and H. Okamoto Angiotensin Type 2 Receptor-Mediated Phosphorylation of eNOS in the Aortas of Mice With 2-Kidney, 1-Clip Hypertension Hypertension, May 1, 2005; 45(5): 967 - 973. [Abstract] [Full Text] [PDF]
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M. Bucci, F. Roviezzo, I. Posadas, J. Yu, L. Parente, W. C. Sessa, L. J. Ignarro, and G. Cirino Endothelial nitric oxide synthase activation is critical for vascular leakage during acute inflammation in vivo PNAS, January 18, 2005; 102(3): 904 - 908. [Abstract] [Full Text] [PDF]
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D. Fulton, M. B. Harris, B. E. Kemp, R. C. Venema, M. B. Marrero, and D. W. Stepp Insulin resistance does not diminish eNOS expression, phosphorylation, or binding to HSP-90 Am J Physiol Heart Circ Physiol, December 1, 2004; 287(6): H2384 - H2393. [Abstract] [Full Text] [PDF]
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M. T. Gentile, C. Vecchione, A. Maffei, A. Aretini, G. Marino, R. Poulet, L. Capobianco, G. Selvetella, and G. Lembo Mechanisms of Soluble {beta}-Amyloid Impairment of Endothelial Function J. Biol. Chem., November 12, 2004; 279(46): 48135 - 48142. [Abstract] [Full Text] [PDF]
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E. K. Walsh, H. Huang, Z. Wang, J. Williams, R. de Crom, R. van Haperen, C. I. Thompson, D. J. Lefer, and T. H. Hintze Control of myocardial oxygen consumption in transgenic mice overexpressing vascular eNOS Am J Physiol Heart Circ Physiol, November 1, 2004; 287(5): H2115 - H2121. [Abstract] [Full Text] [PDF]
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R. Stocker and J. F. Keaney Jr. Role of Oxidative Modifications in Atherosclerosis Physiol Rev, October 1, 2004; 84(4): 1381 - 1478. [Abstract] [Full Text] [PDF]
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M. T Weis, J. L Crumley, L. H Young, and J. N Stallone Inhibiting long chain fatty Acyl CoA synthetase increases basal and agonist-stimulated NO synthesis in endothelium Cardiovasc Res, August 1, 2004; 63(2): 338 - 346. [Abstract] [Full Text] [PDF]
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D. Fulton, R. Babbitt, S. Zoellner, J. Fontana, L. Acevedo, T. J. McCabe, Y. Iwakiri, and W. C. Sessa Targeting of Endothelial Nitric-oxide Synthase to the Cytoplasmic Face of the Golgi Complex or Plasma Membrane Regulates Akt- Versus Calcium-dependent Mechanisms for Nitric Oxide Release J. Biol. Chem., July 16, 2004; 279(29): 30349 - 30357. [Abstract] [Full Text] [PDF]
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J.-P. Gratton, P. Bernatchez, and W. C. Sessa Caveolae and Caveolins in the Cardiovascular System Circ. Res., June 11, 2004; 94(11): 1408 - 1417. [Abstract] [Full Text] [PDF]
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C. D. Fike, J. L. Aschner, Y. Zhang, and M. R. Kaplowitz Impaired NO signaling in small pulmonary arteries of chronically hypoxic newborn piglets Am J Physiol Lung Cell Mol Physiol, June 1, 2004; 286(6): L1244 - L1254. [Abstract] [Full Text] [PDF]
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M. Bucci, F. Roviezzo, V. Brancaleone, M. I. Lin, A. Di Lorenzo, C. Cicala, A. Pinto, W. C. Sessa, S. Farneti, S. Fiorucci, et al. Diabetic Mouse Angiopathy Is Linked to Progressive Sympathetic Receptor Deletion Coupled to an Enhanced Caveolin-1 Expression Arterioscler. Thromb. Vasc. Biol., April 1, 2004; 24(4): 721 - 726. [Abstract] [Full Text] [PDF]
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S. C. Tai, G. B. Robb, and P. A. Marsden Endothelial Nitric Oxide Synthase: A New Paradigm for Gene Regulation in the Injured Blood Vessel Arterioscler. Thromb. Vasc. Biol., March 1, 2004; 24(3): 405 - 412. [Abstract] [Full Text]
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A. Papapetropoulos, D. Fulton, M. I. Lin, J. Fontana, T. J. McCabe, S. Zoellner, G. Garcia-Cardena, Z. Zhou, J.-P. Gratton, and W. C. Sessa Vanadate Is a Potent Activator of Endothelial Nitric-Oxide Synthase: Evidence for the Role of the Serine/Threonine Kinase Akt and the 90-kDa Heat Shock Protein Mol. Pharmacol., February 1, 2004; 65(2): 407 - 415. [Abstract] [Full Text] [PDF]
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K. S. Aulak, T. Koeck, J. W. Crabb, and D. J. Stuehr Dynamics of protein nitration in cells and mitochondria Am J Physiol Heart Circ Physiol, January 1, 2004; 286(1): H30 - H38. [Abstract] [Full Text] [PDF]
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S. Liu, R. T. Premont, C. D. Kontos, J. Huang, and D. C. Rockey Endothelin-1 Activates Endothelial Cell Nitric-oxide Synthase via Heterotrimeric G-protein {beta}{gamma} Subunit Signaling to Protein Kinase B/Akt J. Biol. Chem., December 12, 2003; 278(50): 49929 - 49935. [Abstract] [Full Text] [PDF]
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R. D. Minshall, W. C. Sessa, R. V. Stan, R. G. W. Anderson, and A. B. Malik Caveolin regulation of endothelial function Am J Physiol Lung Cell Mol Physiol, December 1, 2003; 285(6): L1179 - L1183. [Abstract] [Full Text] [PDF]
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M. I. Lin, D. Fulton, R. Babbitt, I. Fleming, R. Busse, K. A. Pritchard Jr., and W. C. Sessa Phosphorylation of Threonine 497 in Endothelial Nitric-oxide Synthase Coordinates the Coupling of L-Arginine Metabolism to Efficient Nitric Oxide Production J. Biol. Chem., November 7, 2003; 278(45): 44719 - 44726. [Abstract] [Full Text] [PDF]
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 R. van Haperen, C. Cheng, B. M. E. Mees, E. van Deel, M. de Waard, L. C.A. van Damme, T. van Gent, T. van Aken, R. Krams, D. J. Duncker, et al. Functional Expression of Endothelial Nitric Oxide Synthase Fused to Green Fluorescent Protein in Transgenic Mice Am. J. Pathol., October 1, 2003; 163(4): 1677 - 1686. [Abstract] [Full Text] [PDF]
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M. Yoshida and Y. Xia Heat Shock Protein 90 as an Endogenous Protein Enhancer of Inducible Nitric-oxide Synthase J. Biol. Chem., September 19, 2003; 278(38): 36953 - 36958. [Abstract] [Full Text] [PDF]
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Y. C. Boo and H. Jo Flow-dependent regulation of endothelial nitric oxide synthase: role of protein kinases Am J Physiol Cell Physiol, September 1, 2003; 285(3): C499 - C508. [Abstract] [Full Text] [PDF]
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H. Hendrickson, S. Chatterjee, S. Cao, M. M. Ruiz, W. C. Sessa, and V. Shah Influence of caveolin on constitutively activated recombinant eNOS: insights into eNOS dysfunction in BDL rat liver Am J Physiol Gastrointest Liver Physiol, August 8, 2003; 285(3): G652 - G660. [Abstract] [Full Text] [PDF]
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Y. Su, S. Edwards-Bennett, M. R. Bubb, and E. R. Block Regulation of endothelial nitric oxide synthase by the actin cytoskeleton Am J Physiol Cell Physiol, June 1, 2003; 284(6): C1542 - C1549. [Abstract] [Full Text] [PDF]
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M. Pelat, C. Dessy, P. Massion, J.-P. Desager, O. Feron, and J.-L. Balligand Rosuvastatin Decreases Caveolin-1 and Improves Nitric Oxide-Dependent Heart Rate and Blood Pressure Variability in Apolipoprotein E-/- Mice In Vivo Circulation, May 20, 2003; 107(19): 2480 - 2486. [Abstract] [Full Text] [PDF]
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S. Wedgwood, C. J. Mitchell, J. R. Fineman, and S. M. Black Developmental differences in the shear stress-induced expression of endothelial NO synthase: changing role of AP-1 Am J Physiol Lung Cell Mol Physiol, April 1, 2003; 284(4): L650 - L662. [Abstract] [Full Text] [PDF]
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