Review
This Review is part of a thematic series on Cyclic GMP–Generating Enzymes and Cyclic GMP–Dependent
Signaling, which includes the following articles:
Regulation of Nitric Oxide–Sensitive Guanylyl Cyclase
Cyclic GMP Phosphodiesterases and Regulation of Smooth Muscle Function
Structure, Regulation, and Function of Mammalian Membrane Guanylyl Cyclase Receptors, With a Focus on
Guanylyl Cyclase-A
Cyclic GMP–Dependent Protein Kinases and the Cardiovascular System: Insights From Genetically Modified Mice
Regulation of Gene Expression by Cyclic GMP
Explaining the Phenomenon of Nitrate Tolerance
Rudi Busse, Editor
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Structure, Regulation, and Function of Mammalian
Membrane Guanylyl Cyclase Receptors, With a Focus on
Guanylyl Cyclase-A
Michaela Kuhn
Abstract—Besides soluble guanylyl cyclase (GC), the receptor for NO, there are at least seven plasma membrane enzymes
that synthesize the second-messenger cGMP. All membrane GCs (GC-A through GC-G) share a basic topology, which
consists of an extracellular ligand binding domain, a short transmembrane region, and an intracellular domain that
contains the catalytic (GC) region. Although the presence of the extracellular domain suggests that all these enzymes
function as receptors, specific ligands have been identified for only three of them (GC-A through GC-C). GC-A
mediates the endocrine effects of atrial and B-type natriuretic peptides regulating arterial blood pressure and volume
homeostasis and also local antihypertrophic actions in the heart. GC-B is a specific receptor for C-type natriuretic
peptide, having more of a paracrine function in vascular regeneration and endochondral ossification. GC-C mediates the
effects of guanylin and uroguanylin on intestinal electrolyte and water transport and on epithelial cell growth and
differentiation. GC-E and GC-F are colocalized within the same photoreceptor cells of the retina and have an important
role in phototransduction. Finally, the functions of GC-D (located in the olfactory neuroepithelium) and GC-G
(expressed in highest amounts in lung, intestine, and skeletal muscle) are completely unknown. This review discusses
the structure and functions of membrane GCs, with special emphasis on the physiological endocrine and cardiac
functions of GC-A, the regulation of hormone-dependent GC-A activity, and the relevance of alterations of the atrial
natriuretic peptide/GC-A system to cardiovascular diseases. (Circ Res. 2003;93:700-709.)
Key Words: guanylyl cyclase receptors
I
n the past 20 years, the purification, cloning, and expression of various forms of guanylyl cyclase (GC) have
revealed that besides soluble GC, the receptor for NO, there
are at least seven mammalian plasma membrane enzymes that
synthesize the second-messenger cGMP. Although partly
homologous to soluble GC, the membrane GCs (GC-A
through GC-G) share a unique topology that consists of an
extracellular ligand binding domain, a short transmembrane
region, and an intracellular domain that contains the catalytic
(GC) region at its C-terminal end (Figure 1). The presence of
the extracellular domain suggests that all these enzymes
function as receptors for specific ligands, but such ligands
have been identified for only three of the membrane forms
(GC-A through GC-C), with the remaining four forms pre-
Original received May 8, 2003; revision received August 27, 2003; accepted August 28, 2003.
From the Institute of Pharmacology and Toxicology, Universitätsklinikum Münster, Münster, Germany.
Correspondence to Dr Michaela Kuhn, Institute of Pharmacology and Toxicology, Universitätsklinikum Münster, Domagkstrasse 12, D-48149
Münster, Germany. E-mail [email protected]
© 2003 American Heart Association, Inc.
Circulation Research is available at http://www.circresaha.org
DOI: 10.1161/01.RES.0000094745.28948.4D
700
Kuhn
Figure 1. Predicted shared basic topology of membrane GC-A
through GC-G. A single transmembrane segment separates an
extracellular domain from intracellular protein kinase–like, hinge,
and cyclase catalytic domains. All membrane forms appear to
form homodimers or higher ordered structures.
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sumed as orphan receptors.1 In contrast to soluble GC
activators, which are small gaseous molecules (NO and CO),
activators of membrane GCs are peptides. Their ligands and
most prominent tissue expression sites are listed in Table 1.
The first mammalian cyclase cDNAs isolated encode specific
receptors for the natriuretic peptide (NP) family: GC-A for
atrial NP (ANP) and B-type NP (BNP)2,3 and GC-B for
C-type NP (CNP).4,5 The structure and regulation of GC-A as
well as the functions of ANP/BNP in blood pressure regulation and cardiac (patho)physiology will be reviewed in the
next sections. CNP is mainly produced by vascular endothelial cells and may act locally as an autocrine/paracrine
regulator of vascular tone and cell growth through GC-B.6 It
potently inhibits the proliferation of vascular smooth muscle
cells but stimulates endothelial growth and might therefore
modulate vascular regeneration.7,8 Surprisingly, targeted disruption of the murine genes for CNP or cGMP-dependent
protein kinase II (PKG II) resulted in severe dwarfism as a
result of impaired endochondral ossification, demonstrating
that the CNP/GC-B system has an essential role in the local
stimulation of growth plate chondrocyte proliferation and
differentiation through cGMP-mediated activation of PKG
II.9,10 GC-C, which contains an extracellular domain with
limited sequence similarity to the above two isoforms, is
mainly expressed in the intestinal epithelium and represents
the receptor for bacterial heat-stable enterotoxins and for two
Membrane Guanylyl Cyclase Receptors
701
endogenous intestinal peptides, guanylin and uroguanylin.11–13 It mediates the local effects of these peptides on
intestinal electrolyte and water transport and epithelial cell
growth and differentiation14 –16 and possibly also the renal
diuretic/natriuretic responses to uroguanylin.17 Additionally,
a role of GC-C in liver regeneration has been suggested.18
However, disruption of the GC-C gene in mice has not
resulted in a unique phenotype.19 Of the four orphan GC
receptors, three are expressed in sensory tissues. GC-E is
expressed in the eye and the pineal gland, whereas GC-F
expression is confined to the retina.20 –22 In the retina, both
cyclases are colocalized within the same photoreceptor cells,
suggesting that they are involved in the phototransduction
cascade.20 Indeed, disruption of the GC-E gene in mice
demonstrated an important role in both the survival of cone
photoreceptors as well as in the maintenance of vision.23 The
expression of GC-D is restricted to a small population of
neurons within a single topographic zone in the olfactory
neuroepithelium; it may function directly in odor recognition
or in modulating the sensitivity of a subpopulation of sensory
neurons to specific odors.24,25 GC-G is the last member of the
membrane GC form to be identified.26 No other mammalian
transmembrane GCs are predicted on the basis of gene
sequence repositories. In contrast to the other orphan receptor
GCs, GC-G has a broad tissue distribution in the rat,
including the lung, intestine, kidney, and skeletal muscle,
raising the possibility that there is another yet-to-bediscovered family of cGMP-generating ligands.26
The intracellular region of all membrane GCs consists of a
juxtamembranous protein kinase– homology domain (KHD),
an amphipathic ␣-helical or hinge region, and a C-terminal
cyclase– homology catalytic domain (Figure 1).27–29 The
function of the KHD is incompletely understood. Although it
binds ATP and contains many residues conserved in the
catalytic domain of protein kinases, kinase activity has not
been detected. It modulates the enzyme activity of the
C-terminal cyclase– homology catalytic domain27,29 –31 and
may act as a docking site for the direct association of
membrane GCs with other proteins, such as protein phosphatase PP5 and HSP90 (GC-A)32 or the intracellular Ca2⫹binding proteins GCAP-1 and GCAP-2 that regulate GC-E
TABLE 1. Prominent Regions of Expression, Ligands, and Main Functions of the Various Mammalian Membrane Guanylyl
Cyclase Receptors
Tissue Distribution
Ligand(s)
Functions
GC-A
Receptor
Vascular smooth muscle, endothelium,
central and peripheral
nervous system, adrenals,
kidney, spleen, heart
ANP, BNP
Decrease in arterial blood pressure
Decrease in blood volume
Inhibition of cardiomyocyte growth
GC-B
Fibroblasts, other tissues
CNP
Endochondral ossification
Vascular regeneration
GC-C
Intestinal epithelium, regenerating liver
Heat-stable enterotoxins, guanylin,
Increased intestinal and renal water and electrolyte transport
uroguanylin
Epithelial cell growth and differentiation
GC-D
Olfactory neuroepithelium
Orphan
Odor recognition?
GC-E
Retina, pineal gland
Orphan
Vision, survival of cones
GC-F
Retina
Orphan
Vision?
GC-G
Skeletal muscle, lung, intestine
Orphan
?
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and GC-F function.33 The coiled-coil hinge region is involved
in higher order oligomerization. Hence, although the transmembrane GC receptors contain a single cyclase catalytic
active site per polypeptide chain, receptor dimerization is
essential for the activation of the catalytic domain.27,34 Presumably, all effects of the membrane GCs are mediated by
the synthesis of cGMP as an intracellular signaling molecule,
which then modulates the activity of specific target proteins,
such as phosphodiesterases, ion channels, and cGMPdependent protein kinases, and thereby modifies cellular
functions (see reviews9,35,36). Last, another common feature
of all membrane GCs, with the exception of GC-F, is the
glycosylation of residues located at the N-terminal end of the
extracellular domain.30 Glycosylation results in heterogeneity
in the size of GC receptors, but the functional consequences
remain controversial. In studies with cells overexpressing
GC-A,37 GC-B,38 or GC-C,39 removal of carbohydrate residues with endoglycosidase prevented or reduced ligand binding, indicating that glycosylation plays a role in ligand
binding. Others instead suggested that glycosylation functions in folding and/or transport of particulate GCs to the cell
membrane.40 Interestingly, a recent report has shown, for the
first time, tissue-specific differences in glycosylation of
GC-A, namely, the occurrence of a specific, less glycosylated
GC-A subtype in the brain; however, the functional implications remain unknown.41
ANP, BNP, and Their Receptor, GC-A
In 1981, de Bold et al42 published a landmark study showing
that rat atrial extracts contain a potent diuretic and natriuretic
factor, thereby establishing for the first time the connection
between the heart and the kidney. This observation led to the
isolation of ANP from cardiac tissue.43 Subsequently, two
other peptides possessing a common core structure (a 17member disulfide ring with a highly conserved internal
sequence of -FGXXXDRIGXXSGL-) have been recognized
and named in alphabetical order, BNP and CNP.44,45 Because
ANP possessed strong natriuretic properties, all three molecules have been referred to as NPs, even though they possess
other activity or may never function to regulate sodium
excretion (ie, CNP; it is found only in traces in blood, and
little evidence suggests that it regulates sodium excretion in
the kidney). ANP and BNP are cardiac hormones that are
produced mainly in the atrium and ventricle, respectively.46
Outside the heart, ANP-specific mRNA or proANP-like
immunoreactivity has been detected in aortic arch cells, lung,
brain, adrenals, kidney, gastrointestinal tract, thymus, choroidea, and ciliary bodies (see comprehensive review47).
However, immunoreactive ANP levels are far lower in these
tissues (⬍1% to 2%) than in cardiac atria; therefore, they are
unlikely to contribute substantially to plasma levels, at least
under physiological conditions. Instead, because most of
these tissues also express GC-A, ANP is likely to exert local
autocrine/paracrine actions. In particular, ANP is synthesized
in several discrete loci within the brain, such as the hypothalamus and the AV3V region, where it may serve as a neuropeptide involved in the modulation of nervous system functions
(see Table 2 and review47). In the kidney, alternative processing of the ANP precursor generates a 32–amino acid peptide
TABLE 2. Hypotensive and Hypovolemic Effects of the
ANP/GC-A System
Hormonal Effects
Inhibition of the renin-angiotensin-system61
Fall in aldosterone and cortisol and rise in testosterone levels47,62
Renal Effects
Rise in glomerular filtration rate63
Rise in filtration fraction64
Rise in renovascular resistance65
Inhibition of sodium reabsorption in the inner medullary collecting duct66
Increase in urinary volume and electrolytes60
Effects in the Spleen
Increased splenic microvascular pressure67
Fluid extravasation from the splenic vasculature into the lymphatic
system67
Vascular Effects
Vasodilatation68
Modulation of endothelial permeability69
Central Nervous System Effects
Modulation of sympathetic activity70,71
Inhibition of arginine vasopressin secretion47,72
Diminished salt appetite and water drinking47,60
Effects on blood pressure/volume-regulating regions in the brain47,60
called urodilatin, which may be important for the local
regulation of renal sodium and water handling.48
The main triggering factor for the cardiac release/production of ANP and BNP is an increase in wall stretch and/or
pressure,46 but neurohumoral factors such as glucocorticoids,
catecholamines, arginine vasopressin, angiotensin II (Ang II),
and endothelin may also play a role.47 Cardiac ANP and BNP
are released into the bloodstream, activate the GC-A receptor,
which is expressed in a variety of tissues, and thereby
modulate blood pressure/volume (see Table 2). Competitive
binding against 125I-human ANP on human GC-A established
a higher potency of human ANP compared with human BNP,
with dissociation constants (Kds) in the range of 1.6 pM for
ANP and 7.3 pM for BNP.49 The plasma half-life of ANP and
BNP in humans is 2 to 5 minutes, and elimination occurs by
enzyme metabolism, by cellular internalization by specific
clearance receptors (NP receptor-C [NPR-C]; see next section), and (to a limited extend) by urinary excretion. The
degrading enzyme is a neutral endopeptidase (NEP 24.11)
maximally found in the brush border of the proximal convoluted tubule and also found in the lungs, intestine, seminal
vesicles, and neutrophiles.50
The important role of the ANP/GC-A system in the
physiological regulation of arterial blood pressure/volume
has been emphasized in different genetic mouse models.
Targeted deletion of the peptide (ANP⫺/⫺) or its receptor
(GC-A⫺/⫺), leads to severe, chronic arterial hypertension,
cardiac hypertrophy, and sudden death51–54 (Figure 2). In
contrast, overexpression of ANP or GC-A elicits a “dosedependent” fall in arterial blood pressure.55,56 Intriguingly,
although BNP and ANP appear to signal through the same
receptor, mice without BNP exhibit a different phenotype
Kuhn
Figure 2. Targeted global deletion of the GC-A gene in mice
(GC-A⫺/⫺) leads to severe, chronic arterial hypertension and
marked cardiac hypertrophy. *P⬍0.05 vs corresponding wildtype (GC-A⫹/⫹) mice. Data from Kuhn et al.78
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than do ANP-deficient mice. Whereas BNP-deficient mice do
not have hypertension or cardiac hypertrophy, they are
susceptible to cardiac fibrosis.57 Thus, gene-deletion experiments suggest that ANP and BNP have distinct physiological
roles. Under physiological conditions (in the absence of
cardiac pressure or volume overload), the peripheral circulating plasma concentrations of BNP are much lower than the
concentrations of ANP. Also, as mentioned above, the affinity of BNP for binding to GC-A is lower than that for ANP.49
Accordingly, the potency for vasorelaxation is also markedly
less.58 Thus, it is possible that BNP (which is constitutively
expressed in cardiomyocytes) mainly acts as a local paracrine
antifibrotic factor within the heart (ie, because fibroblasts
express relatively high levels of both GC-A and GC-B).
However, the following observations suggest that BNP might
activate another so-far-unknown receptor: (1) Whereas the
amino acid sequences of ANP and CNP are conserved among
animal species, the sequence of BNP is highly different
between humans, rats, and mice. (2) Some tissues of GC-A⫺/⫺
mice (ie, the testis and adrenal gland) retain significant
high-affinity cGMP responses to BNP. This residual response
cannot be accounted for by GC-B or any other known
mammalian membrane GC,59 suggesting that an as-yetunidentified receptor may exist that specifically recognizes
BNP.
Endocrine Actions of the ANP/GC-A System
Cooperate in Acute and Chronic Regulation of
Blood Pressure and Blood Volume
As depicted in Table 1, the GC-A receptor is expressed in
many different organs and cell types and mediates a variety of
central and peripheral actions of ANP, which probably all
contribute to lower blood pressure (see Table 2).47,60 –72 In
particular, the renal63– 66 and sympatholytic actions of
ANP70,71 probably have a major role in the hypotensive
actions of the peptide. In contrast, the relevance of the direct
vasodilating ANP effect is controversial, because very variable GC-A levels are expressed in different vascular beds.68,73
To characterize this issue, in a recent study, we selectively
deleted GC-A in vascular smooth muscle cells (SMC GC-A
KO) using Cre-lox technology.74 Surprisingly, in spite of the
clear abolition of the direct vasorelaxing effects of ANP, the
resting arterial blood pressure of conscious SMC GC-A KO
mice was completely normal (Figure 3A). However, these
Membrane Guanylyl Cyclase Receptors
703
Figure 3. Effect of selective deletion of GC-A in vascular
smooth muscle cells (SMC GC-A KO mice) on arterial blood
pressure (Cre-lox technology). A, Resting arterial blood pressure
of conscious SMC GC-A KO mice was not different from floxed
GC-A control mice. B, Changes in systolic blood pressure
(⌬mm Hg) during and immediately after acute volume expansion
are compared with baseline values. Whereas floxed GC-A control mice are totally able to compensate this volume overload,
SMC GC-A KO mice react with a prompt and marked hypertension. *P⬍0.05 vs baseline, before volume overload; †P⬍0.05 vs
floxed GC-A. Modified from Holtwick R, Gotthardt M, Skryabin
B, Steinmetz M, Potthast R, Zetsche B, Hammer B, Herz J,
Kuhn M. Smooth muscle-selective deletion of guahylyl
cyclase-A prevents the acute but not chronic effects of ANP on
blood pressure. Proc Natl Acad Sci U S A. 2002;99:7142–7147.
Copyright 2002, National Academy of Sciences U.S.A.
mice reacted to acute vascular volume expansion with a
prompt and marked hypertension, which was never observed
in control mice expressing normal GC-A levels (Figure 3B).74
On the basis of these observations, we suggest that vascular
GC-A is dispensable in the chronic but crucial in the acute
moderation of blood pressure by ANP.
Local Cardiac Functions of the ANP/GC-A System
During chronic hemodynamic overload, the expression levels
of ANP and especially BNP in the cardiac ventricles significantly increase.46 NPs in this situation may act not only as
circulating endocrine factors to maintain arterial blood pressure and volume homeostasis but also as local antihypertrophic (ANP) and antifibrotic (BNP) cardiac factors.57 For
example, ANP inhibits growth and proliferation of cultured
cardiac myocytes and fibroblasts via GC-A.75,76 Also, targeted overexpression of GC-A in cardiomyocytes exerts
antihypertrophic effects in vivo.77 Conversely, mice with a
global genetic disruption of the GC-A gene (GC-A⫺/⫺ mice)
not only have increased systemic blood pressure but also
display a marked cardiac hypertrophy that is disproportionate
to their increased blood pressure and resistant to antihypertensive medication.52,53,78 To test whether ANP locally modulates cardiomyocyte growth and contractility in vivo, we
generated mice with selective deletion of GC-A in cardiomyocytes.79 All endocrine blood pressure and volumeregulating effects of ANP are preserved. Our studies in these
mice demonstrate that the ANP/GC-A system indeed plays an
autocrine/paracrine role in the heart, which inhibits cardiomyocyte growth and stimulates diastolic relaxation.79 Furthermore, our results corroborate published studies indicating
that ANP inhibits its own synthesis and release via GC-A/
cGMP as a negative-feedback mechanism (see Figure 4).79
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Figure 4. Effect of selective deletion of GC-A in cardiomyocytes
(CM GC-A KO mice) on systolic blood pressure (A), the heart
weight (HW) to body weight (BW) ratios (B), and the circulating
plasma concentrations of ANP (C) and cGMP (D). CM GC-A KO
mice exhibit arterial hypotension and mild cardiac hypertrophy.
This is accompanied by elevated plasma levels of ANP and
cGMP, indicating enhanced cardiac release and increased peripheral effects of the peptide. The latter probably contributes to
the hypotensive phenotype. *P⬍0.05 vs corresponding floxed
GC-A control mice. Panels A, B, and C are from Holtwick et
al.79 (J Clin Invest. 2003;111:1399 –1407), with permission.
In summary, these observations from various genetic
mouse models with global or conditional cell-restricted deletion of GC-A emphasize the importance of the NP/GC-A/
cGMP system in the endocrine acute and chronic regulation
of arterial blood pressure and blood volume and also its local
actions on cardiac growth, remodeling, and contractile
functions.
Regulation of ANP/BNP-Dependent
GC-A Activity
In some forms of arterial hypertension and as one of the
earliest and pathognomonic events in cardiac hypertrophy
and insufficiency, the cardiac synthesis and release of ANP
and BNP is markedly enhanced, but the GC-A–mediated
effects of the NPs are clearly diminished, indicating a
receptor or postreceptor defect of GC-A (see next section).
Thus, from a clinical and pathophysiological perspective,
identifying the specific biochemical and genetic mechanisms
involved in the downregulation of GC-A activity has important implications. The present understanding of the regulation
of hormone-dependent GC-A activity, derived mainly from in
vitro studies using cultured cell lines, is incomplete. Five
different mechanisms have been implicated: (1) sequestration
of NPs by the clearance receptor (NPR-C), (2) phosphorylation and dephosphorylation of GC-A, (3) receptor internalization, (4) regulation of GC-A signaling activity by extracellular tonicity, and (5) regulation of GC-A at the level of
gene transcription.
Sequestration of NPs by NPR-C
Besides GC-A and GC-B, a third specific receptor subtype
which triggers the effects of NPs is the C-receptor (NPRC),60,80 a clearance receptor that mainly serves for cellular
internalization and degradation of NPs. In many tissues,
Figure 5. Hypothetical model of the process of ANP- and BNPdependent activation and homologous desensitization of GC-A
and the role of receptor phosphorylation and dephosphorylation.
P indicates phosphate.
NPR-C is the most abundant of the NP receptors, and it binds
ANP, BNP, and CNP with relatively similar affinities.80 The
extracellular domain shares a high amino acid sequence
homology with GC-A and GC-B. However, it has only 37
intracellular amino acids and does not possess GC activity.80
It may participate in mediating some of the cellular actions of
NPs via coupling to Gi proteins and negative modulation of
adenylyl cyclase activity.81 However, within the cardiovascular system, the primary action of the NPR-C seems to be
the modulation of circulating and local NP concentrations
that are available to bind GC-A and GC-B.82 Hence, changes
in GC-A–mediated responses to ANP could theoretically
result from changes in the expression levels of NPR-C.
Indeed, an upregulation of NPR-C has been described in at
least two clinical settings with blunted responses to exogenous ANP: (1) in obesity-related hypertension83 and (2) in
patients with chronic heart failure84 (see next section). Vice
versa, it is conceivable that a downregulation of NPR-C
enhances the effects of ANP under specific physiological
conditions. For instance, continued exposure of cultured
endothelial cells to increased NaCl concentrations resulted in
a dramatic loss of NPR-C, which was accompanied with a
marked potentiation of ANP/GC-A–induced cGMP accumulation.85 Although not confirmed in vivo, these observations
raise the possibility that changes in extracellular tonicity, in
particular, in the kidney, could modulate NPR-C expression
levels and thereby the interaction of NPs with GC-A.
Phosphorylation and Dephosphorylation of GC-A
Biochemical studies in transfected GC-A– overexpressing
cells showed that phosphorylation of GC-A within the KHD
is essential for its activation process.86 – 88 In turn, desensitization and/or inactivation of GC-A probably involves ANPdependent dephosphorylation of the KHD.86 On these experiments, the groups of Garbers (Foster et al89) and Potter
(Potter and Hunter90) constructed a working model of the
process of ANP- and BNP-dependent activation and deactivation of GC-A, which is illustrated in Figure 5 (see comprehensive reviews89,90). In the absence of NP, the receptor is
highly phosphorylated on at least six amino acid residues
located within the KHD, and its GC activity is repressed. On
NP binding, a conformational change occurs that facilitates
Kuhn
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three subsequent events. First, the normal inhibitory effect
that the KHD has on catalytic activity is relieved. Second, an
increased dissociation rate decreases the affinity of the
extracellular domain of GC-A for ANP and BNP.91 Third, a
conformational change in the KHD may expose the phosphorylated residues to a specific protein phosphatase. The
resulting dephosphorylated receptor is unresponsive to further hormonal stimulation. Unfortunately, the specific kinase(s) and phosphatase(s) involved in the phosphorylation
(sensitization) and dephosphorylation (desensitization) of
GC-A remain unknown. Notably, a very recent study suggested that cGMP-dependent protein kinase type I (PKG I)
might be involved.92 The authors showed that ANP stimulation of GC-A recruits PKG I to the plasma membrane and
simultaneously promotes PKG I activation via ANP/GC-A–
generated cGMP. Hence, an ANP-stimulated GC-A–PKG
association may represent a novel mechanism for both compartmentation of cGMP-mediated signaling and regulation of
receptor sensitivity.92
NP-dependent activation of GC-A can be reduced not only
by chronic exposure to NPs (homologous desensitization) but
also by exposure to agents other than NPs (heterologous
desensitization). It has been shown in cell culture systems that
Ang II and endothelin decrease the responsiveness of GCA.92,93 This is probably mediated by a protein kinase C–induced dephosphorylation of GC-A.93–95 Whether the protein
kinase C– dependent dephosphorylation of GC-A results from
activation of a protein phosphatase or the inhibition of a
protein kinase is not known. However, on the basis of these
observations, it is conceivable that increased local concentrations of endothelin, Ang II, or other growth factors such as
platelet-derived growth factor and fibroblast growth factor96
interfere with NP/GC-A signaling under certain pathological
conditions in vivo.
Receptor Internalization
Another process that may account for ANP-dependent downregulation of GC-A is the internalization of ligand-receptor
complexes. So far, this phenomenon has been shown only in
stably transfected 293 cells expressing a very high density of
recombinant GC-A receptors.97 Most of the internalized ANP
and GC-A was degraded. But a portion of the internalized
ligand-receptor complexes dissociated intracellularly, escaping the lysosomal degradative pathway and recycling back to
the plasma membrane.97
Regulation of GC-A Signaling Activity by
Extracellular Tonicity
Studies in various cultured cell types have shown that
hyperosmolarity acutely inhibits GC-A,98,99 whereas chronic
exposure results in elevated GC-A activity.100,101 Chen and
Gardner101 demonstrated in primary cultures of rat inner
medullary collecting duct cells that both the expression and
cGMP-synthesizing activity of GC-A are stimulated by increased extracellular tonicity and that this process seems to
involve the activity of p38 mitogen-activated protein kinase.
Thus, changes in extracellular tonicity, in particular in the
kidney, might modulate the diuretic/natriuretic responses to
Membrane Guanylyl Cyclase Receptors
705
ANP at the level of the expression and also the sensitivity of
both GC-A and NPR-C.
Regulation of GC-A at the Level of
Gene Transcription
In addition to these short-term posttranscriptional mechanisms, regulation of GC-A gene transcription also plays a role
in the regulation of receptor activity. Several studies have
shown that GC-A mRNA and receptor number are reduced in
cultured cells stimulated for a prolonged period of time with
ANP.102 Functional analysis of the GC-A gene promoter
revealed a putative cGMP-responsive element, suggesting
that the ANP-dependent downregulation of GC-A mRNA is
cGMP dependent.103,104 However, other studies could not
reproduce this result.105
Two recent in vitro studies have indicated that GC-A gene
transcription might be inhibited by Ang II and endothelin.106,107 It has been suggested that local endothelin production mediates the effects of changes in extracellular tonicity
on GC-A expression in the kidney.107 Again, the relevance of
these experiments to the in vivo situation needs to be
confirmed.
In summary, the regulation of hormone-dependent GC-A
activity is complex and surely involves different processes at
both the posttranscriptional and transcriptional level. Which
of these processes accounts for the termination and physiological as well as pathophysiological modulation of effects of
NP in vivo remains an open and important question. It is also
likely that different types of tissues and cells rely on different
mechanisms to regulate GC-A activity.
Relevance of Alterations in the ANP/GC-A
System to Cardiovascular Diseases
The findings summarized in the present review indicate that
the ANP/GC-A system is not only critical in the regulation of
blood pressure/volume but also locally involved in moderating the cardiac growth response to hypertrophic stimuli,
suggesting that some forms of hypertension and/or cardiac
hypertrophy in humans are explained in part by inappropriate
secretion of ANP or diminished expression and/or responsiveness of GC-A.
Quantitative alterations in gene expression governed by
polymorphisms in noncoding sequences seem to contribute to
the genetic susceptibility to complex diseases, such as essential hypertension (EH) and cardiac hypertrophy. Many of
such variations have been published within the human genes
for ANP, GC-A, and NPR-C, and some interesting examples
will be mentioned here. An HpaII-polymorphism in intron 2
of the ANP gene is more frequently observed in salt-sensitive
hypertensive African Americans (50%) compared with normotensive subjects (3%) or with white patients with EH
(15%).108,109 For GC-A, the association between a functional
deletion mutation in the promoter region of the human GC-A
gene, decreased receptor expression, and EH as well as
ventricular hypertrophy has been shown in the Japanese
population110 but not in European patients with EH.111
Knowles et al112 recently identified 10 additional polymorphic sites in the noncoding sequence of GC-A and, by
transient expression analysis, demonstrated that they can alter
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GC-A expression as much as 2-fold. Last, a promoter variant
of the NPR-C gene has been shown to be associated with
lower ANP levels and higher blood pressure in obese
hypertensives.113
Apart from these genetic variations, functional alterations
of ANP or GC-A might also be involved in some forms of
EH. Inappropriate ANP secretion was reported in saltsensitive hypertensive black patients manifesting a paradoxical decrease in ANP secretion under conditions of high salt
intake.114 On the other hand, blunted vasodilating and diuretic/natriuretic responses to exogenous ANP have been reported in EH,111 in Cushing’s disease,115 and in hypertensive
obese patients.83 The latter group has been shown to exhibit
a lower GC-A/NPR-C mRNA ratio in adipose tissue as well
as diminished plasma ANP levels, suggesting enhanced
sequestration and clearance of ANP by NPR-C in adipocytes.
Patients with cardiac hypertrophy and/or congestive heart
failure (CHF) have elevated plasma levels of ANP and BNP,
with these peptide levels being highly related to the severity
of the disease.116 However, the cardiovascular and cGMP
responses to exogenous ANP or BNP are markedly attenuated, indicating a downregulation or impaired receptor or
postreceptor responsiveness of GC-A (as demonstrated by
Hirooka et al117 and many others). Some studies have
suggested that the decreased biological responses to ANP are
partly due to an upregulation of NPR-C receptors in peripheral tissues, leading to increased sequestration and degradation of ANP.84 Other studies have even indicated a downregulation of NPR-C receptors.118 Thus, the mechanisms
accounting for a diminished NP-dependent stimulation of
GC-A in patients with severe CHF are poorly understood.
The aforementioned processes of downregulation and/or homologous/heterologous desensitization of GC-A may be involved. Using various methodologies, such as “real-time”
quantitative reverse transcription–polymerase chain reaction
and stimulation of cGMP production, we recently determined
the level of transcripts of both GC-A and NPR-C as well as
ANP-stimulated GC-A activity in explanted hearts from CHF
patients subjected to cardiac transplantation. Compared with
“nonfailing” hearts, the cardiac GC-A/NPR-C mRNA ratios
and ANP-dependent GC-A activities were significantly decreased in both ischemic and dilated cardiomyopathy (author’s unpublished observations, 2003). Our studies in mice
with cardiomyocyte-restricted deletion of GC-A indicate that
an inhibition of the local cardiac ANP effects might accelerate the progression of cardiac hypertrophy and insufficiency79
and support further work to assess the importance of the
ANP/GC-A system in human cardiovascular diseases and, in
particular, its systemic but also local cardiac alterations.
Future Directions
The findings described in the present review open several
new directions for study. First, they demonstrate the potential
utility and power of the Cre-lox approach in dissecting out the
tissue-specific functions of ANP and GC-A. This is important
because the GC-A receptor is widely distributed in many
different cell types and mediates the effects of ANP and BNP
in many biological functions. In addition to its role in blood
pressure control, recent reports have indicated that GC-A
might be critical in the regulation of other completely
different physiological processes, such as cellular growth in
the brain and kidney,60 angiogenesis,119 liver regeneration,120
or even lipolysis in adipose tissue.121
Second, an important and still unresolved issue is whether
cGMP production by different membrane-bound versus soluble GCs is compartmentalized. For instance, it has been
shown that GC-A, but not soluble GC, has potent effects on
plasma membrane control of the calcium ATPase pump.122
Interestingly, a very recent study has demonstrated that
ANP/GC-A, but not NO/soluble GC, stimulates the translocation of PKG I to the plasma membrane.92 Hence, the
mechanisms for compartmentation of cGMP-mediated signaling within different cell types remain an intriguing issue.
Third, the present review also demonstrates that the processes regulating GC-A activity and responsiveness in vivo
are largely unknown. In view of the many clinical studies
showing that in some forms of hypertension and in all forms
of cardiac insufficiency the GC-A–mediated effects of ANP
and BNP are greatly diminished, clarification of the responsible GC-A receptor or postreceptor defects and identification
of proteins that regulate the activity of GC-A may have
important pathophysiological implications. This could lead to
the development of drugs that could revolutionize the treatment of some forms of cardiac disease. Even more, because
synthetic BNP (Nesiritide) and ANP (Anaritide) have been
proven to be clinically highly beneficial in the acute treatment
of CHF,123 an understanding of the mechanisms involved in
the ligand-dependent desensitization of GC-A might also be
important from this therapeutical perspective.
As a final point, additional work may uncover other
receptor GCs. Although seven have been identified in mammals, 29 genes encode putative GCs in Caenorhabditis
elegans, even though the nematode genome is only 1/30 that
of the mammalian genome.124 Clearly, there is an exciting
field of physiological and putative clinical importance with
more questions than answers.
Acknowledgments
The author’s work reviewed in the present article was supported by
grants from the Bundesministerium für Bildung und Forschung
(BMBF 01EC9801), the University of Münster (Interdisziplinäre
Klinische Forschung, IZKF B12), and the Deutsche Forschungsgemeinschaft (SFB 556, DFG 1037/3-1). The author is deeply grateful
to Dr Lawrence A. Scheving, Department of Pediatrics at Vanderbilt
University, Nashville, Tenn, for his great help in preparing the
manuscript.
References
1. Garbers DL, Lowe DG. Guanylyl cyclase receptors. J Biol Chem.
1994;269:30741–30744.
2. Chinkers M, Garbers DL, Chang M-S, Lowe DG, Chin H, Goeddel DV,
Schulz S. A membrane form of guanylate cyclase is an atrial natriuretic
peptide receptor. Nature. 1989;338:78 – 83.
3. Lowe DG, Chang M-S, Hellmiss R, Chen E, Singh S, Garbers DL,
Goeddel DV. Human atrial natriuretic peptide receptor defines a new
paradigm for second messenger signal transduction. EMBO J. 1989;8:
1377–1384.
4. Schulz S, Singh S, Bellet RA, Singh G, Tubb DJ, Chin H, Garbers DL.
The primary structure of a plasma membrane guanylate cyclase demonstrates diversity within this new receptor family. Cell. 1989;58:
1155–1162.
Kuhn
Downloaded from http://circres.ahajournals.org/ by guest on July 31, 2017
5. Koller KJ, Lowe DG, Bennett GL, Minamino N, Kangawa K, Matsuo H,
Goeddel DV. Selective activation of the B natriuretic peptide receptor by
C-type natriuretic peptide (CNP). Science. 1991;252:120 –123.
6. Chen HH, Burnett JC Jr. C-type natriuretic peptide: the endothelial
component of the natriuretic peptide system. J Cardiovasc Pharmacol.
1998;32:S22–S28.
7. Komatsu Y, Itoh H, Suga S-I, Ogawa Y, Hama N, Kishimoto I,
Nakagawa O, Igaki T, Doi K, Yoshimasa T, Nakao K. Regulation of
endothelial production of C-type natriuretic peptide in coculture with
vascular smooth muscle cells: role of the vascular natriuretic peptide
system in vascular growth inhibition. Circ Res. 1996;78:606 – 614.
8. Yamahara K, Itoh H, Chun TH, Ogawa Y, Yamashita J, Sawada N,
Fukunaga Y, Sone M, Yurugi-Kobayashi T, Miyashita K, Tsujimoto H,
Kook H, Feil R, Garbers DL, Hofmann F, Nakao K. Significance and
therapeutic potential of the natriuretic peptides/cGMP/cGMP-dependent
protein kinase pathway in vascular regeneration. Proc Natl Acad Sci
U S A. 2003;100:3404 –3409.
9. Pfeifer A, Ruth P, Dostmann W, Sausbier M, Klatt P, Hofmann F.
Structure and function of cGMP-dependent protein kinases. Rev Physiol
Biochem Pharmacol. 1999;135:105–149.
10. Chusho H, Tamura N, Ogawa Y, Yasoda A, Suda M, Miyazawa T,
Nakamura K, Nakao K, Kurihara T, Komatsu Y, Itoh H, Tanaka K, Saito
Y, Katsuki M, Nakao K. Dwarfism and early death in mice lacking
C-type natriuretic peptide: regeneration. Proc Natl Acad Sci U S A.
2001;98:4016 – 4021.
11. Schulz S, Green CK, Yuen PS, Garbers DL. Guanylyl cyclase is a
heat-stable enterotoxin receptor. Cell. 1990;63:941–948.
12. Currie MG, Fok KF, Kato J, Moore RJ, Hamra FK, Duffin KL, Smith
CE. Guanylin: an endogenous activator of intestinal guanylate cyclase.
Proc Natl Acad Sci U S A. 1992;89:947–951.
13. Hamra FK, Forte LR, Eber SL, Pidhorodeckyj NV, Krause WJ, Freeman
RH, Chin DT, Tompkins JA, Fok KF, Smith CE. Uroguanylin: structure
and activity of a second endogenous peptide that stimulates intestinal
guanylate cyclase. Proc Natl Acad Sci U S A. 1993;90:10464 –10468.
14. Forte LR, Currie MG. Guanylin: a peptide regulator of epithelial
transport. FASEB J. 1995;9:643– 650.
15. Kuhn M, Adermann K, Jähne J, Forssmann W-G, Rechkemmer G.
Segmental differences in the effects of guanylin and Escherichia coli
heat-stable enterotoxin on electrogenic chloride secretion in human
intestine. J Physiol (Lond). 1994;479:433– 440.
16. Steinbrecher KA, Wowk SA, Rudolph JA, Witte DP, Cohen MB.
Targeted inactivation of the mouse guanylin gene results in altered
dynamics of colonic epithelial proliferation. Am J Pathol. 2002;161:
2169 –2178.
17. Forte LR, London RM, Freeman RH, Krause WJ. Guanylin peptides:
renal actions mediated by cyclic GMP. Am J Physiol. 2000;278:
F180 –F191.
18. Scheving LA, Russell WE. Guanylyl cyclase C is up-regulated by
nonparenchymal cells and hepatocytes in regenerating rat liver. Cancer
Res. 1996;56:5186 –5191.
19. Schulz S, Lopez MJ, Kuhn M, Garbers DL. Disruption of the guanylyl
cyclase-C gene leads to a paradoxical phenotype of viable but heatstable enterotoxin-resistant mice. J Clin Invest. 1997;100:1590 –1595.
20. Yang R-B, Foster DC, Garbers DL, Fülle H-J. Two membrane forms of
guanylyl cyclase found in the eye. Proc Natl Acad Sci U S A. 1995;92:
602– 606.
21. Shyjan AW, de Sauvage FJ, Gillett NA, Goeddel DV, Lowe DG.
Molecular cloning of a retina-specific membrane guanylyl cyclase.
Neuron. 1992;9:727–737.
22. Lowe DG, Dizhoor AM, Liu K, Gu Q, Spencer M, Laura R, Lu L,
Hurley JB. Cloning and expression of a second photoreceptor-specific
membrane retina guanylyl cyclase (RetGC), RetGC-2. Proc Natl Acad
Sci U S A. 1995;92:5535–5539.
23. Yang R-B, Robinson SW, Xiong WH, Yau KW, Birch DG, Garbers DL.
Disruption of a retinal guanylyl cyclase gene leads to cone-specific
dystrophy and paradoxical rod behavior. J Neurosci. 1999;19:
5889 –5897.
24. Fulle HJ, Vassar R, Foster DC, Yang R-B, Axel R, Garbers DL. A
receptor guanylyl cyclase expressed specifically in olfactory sensory
neurons. Proc Natl Acad Sci U S A. 1995;92:3571–3575.
25. Julifs DM, Fülle H-J, Zhao AZ, Houslay MD, Garbers DL, Beavo JA. A
subset of olfactory neurons that selectively express cGMP-stimulated
phosphodiesterase (PDE2) and guanylyl cyclase-D define a unique
olfactory signal transduction pathway. Proc Natl Acad Sci U S A. 1997;
94:3388 –3395.
Membrane Guanylyl Cyclase Receptors
707
26. Schulz S, Wedel BJ, Matthews A, Garbers DL. The cloning and
expression of a new guanylyl cyclase orphan receptor. J Biol Chem.
1998;273:1032–1037.
27. Wilson EM, Chinkers M. Identification of sequences mediating guanylyl
cyclase dimerization. Biochemistry. 1995;34:4696 – 4701.
28. Liu Y, Ruoho AE, Rao VD, Hurley JH. Catalytic mechanism of the
adenylyl and guanylyl cyclases: modeling and mutational analysis. Proc
Natl Acad Sci U S A. 1997;94:13414 –13419.
29. Chinkers M, Garbers DL. The protein kinase domain of the ANP
receptor is required for signaling. Science. 1989;245:1392–1394.
30. Garbers DL. Guanylyl cyclase-linked receptors. Pharmacol Ther. 1991;
50:337–345.
31. Koller KJ, de Sauvage FJ, Lowe DG, Goeddel DV. Conservation of the
kinaselike regulatory domain is essential for activation of the natriuretic
peptide receptor guanylyl cyclases. Mol Cell Biol. 1992;12:2581–2590.
32. Kumar R, Grammatikakis N, Chinkers M. Regulation of the atrial
natriuretic peptide receptor by heat shock protein 90 complexes. J Biol
Chem. 2001;276:11371–11375.
33. Laura RP, Hurley JB. The kinase homology domain of retinal guanylyl
cyclases 1 and 2 specifies the affinity and cooperativity of interaction
with guanylyl cyclase activating protein-2. Biochemistry. 1998;37:
11264 –11271.
34. Yang R-B, Garbers DL. Two eye guanylyl cyclases are expressed in the
same photoreceptor cells and form homomers in preference to heteromers. J Biol Chem. 1997;272:13738 –13742.
35. Soderling SH, Beavo JA. Regulation of cAMP and cGMP signaling:
new phosphodiesterases and new functions. Curr Opin Cell Biol. 2000;
12:174 –179.
36. Lohmann SM, Vaandrager AB, Smolenski A, Walter U, De Jonge HR.
Distinct and specific functions of cGMP-dependent protein kinases.
Trends Biochem Sci. 1997;22:307–312.
37. Lowe DG, Fendly BM. Human natriuretic peptide receptor-A guanylyl
cyclase: hormone cross-linking and antibody reactivity distinguish
receptor glycoforms. J Biol Chem. 1992;267:21691–21697.
38. Fenrick R, Bouchard N, McNicoll N, De Lean A. Glycosylation of
asparagine 24 of the natriuretic peptide receptor-B is crucial for the
formation of a competent ligand binding domain. Mol Cell Biochem.
1997;173:25–32.
39. Hasegawa M, Hidaka Y, Wada A, Hirayama T, Simonishi Y. The
relevance of N-glycosylation to the binding of a ligand to guanylate
cyclase C. Eur J Biochem. 1999;263:338 –346.
40. Miyagi M, Zhang X, Misono KS. Glycosylation sites in the atrial
natriuretic peptide receptor: oligosaccharide structures are not required
for hormone binding. Eur J Biochem. 2000;267:5758 –5768.
41. Müller D, Middendorff R, Olcese J, Mukhopadhyay AK. Central
nervous system-specific glycosylation of the type A natriuretic peptide
receptor. Endocrinology. 2002;143:23–29.
42. de Bold AJ, Borenstein HB, Veress AT, Sonnenberg H. A rapid and
potent natriuretic response to intravenous injection of atrial myocardial
extract in rats. Life Sci. 1981;28:89 –94.
43. Kangawa K, Matsuo H. Purification and complete amino acid sequence
of ␣-human atrial natriuretic polypeptide (␣-hANP). Biochem Biophys
Res Commun. 1984;118:131–139.
44. Sudoh T, Kangawa K, Minamino N, Matsuo H. A new natriuretic
peptide in porcine brain. Nature. 1988;332:78 – 81.
45. Sudoh T, Minamino N, Kangawa K, Matsuo H. C-type natriuretic
peptide (CNP): a new member of natriuretic peptide family identified in
porcine brain. Biochem Biophys Res Commun. 1990;168:863– 870.
46. de Bold AJ, Ma KK, Zhang Y, de Bold ML, Bensimon M, Khoshbaten
A. The physiological and pathophysiological modulation of the
endocrine function of the heart. Can J Physiol Pharmacol. 2001;79:
705–714.
47. Ruskoaho H. Atrial natriuretic peptide: synthesis, release, and metabolism. Pharmacol Rev. 1992;44:481– 602.
48. Schulz-Knappe P, Forssmann K, Herbst F, Hock D, Pipkorn R,
Forssmann WG. Isolation and structural analysis of “urodilatin,” a new
peptide of the cardiodilatin-(ANP)-family, extracted from human urine.
Klin Wochenschr. 1988;66:752–759.
49. Bennett, BD, Bennett GL, Vitangcol RV, Jewett JRS, Burnier J, Henzel
W, Lowe DG. Extracellular domain-IgG fusion proteins for three human
natriuretic peptide receptors. J Biol Chem. 1991;266:23060 –23067.
50. Matsumura T, Kugiyama K, Sugiyama S, Ohgushi M, Amanaka K,
Suzuki M, Yasue H. Neutral endopeptidase 24.11 in neutrophils modulates protective effects of natriuretic peptides against neutrophilinduced endothelial cytotoxicity. J Clin Invest. 1996;97:2192–2203.
708
Circulation Research
October 17, 2003
Downloaded from http://circres.ahajournals.org/ by guest on July 31, 2017
51. John SW, Veress AT, Honrath U, Chong CK, Peng L, Smithies O,
Sonnenberg H. Blood pressure and fluid-electrolyte balance in mice with
reduced or absent ANP. Am J Physiol. 1996;271:R109 –R114.
52. Lopez MJ, Wong SK, Kishimoto I, Dubois S, Mach V, Friesen J,
Garbers DL, Beuve A. Salt-resistant hypertension in mice lacking the
guanylyl cyclase-A receptor for atrial natriuretic peptide. Nature. 1995;
378:65– 68.
53. Oliver PM, Fox JE, Kim R, Rockman HA, Kim HS, Reddick RL,
Pandey KN, Milgram SL, Smithies O, Maeda N. Hypertension, cardiac
hypertrophy, and sudden death in mice lacking natriuretic peptide
receptor A. Proc Natl Acad Sci U S A. 1997;94:14730 –14735.
54. Lopez MJ, Garbers DL, Kuhn M. The guanylyl cyclase-deficient mouse
defines differential pathways of natriuretic peptide signaling. J Biol
Chem. 1997;272:23064 –23068.
55. Steinhelper ME, Cochrane KL, Field LJ. Hypotension in transgenic mice
expressing atrial natriuretic factor fusion genes. Hypertension. 1990;16:
301–307.
56. Oliver PM, John SW, Purdy KE, Kim R, Maeda N, Goy MF, Smithies
O. Natriuretic peptide receptor 1 expression influences blood pressures
of mice in a dose-dependent manner. Proc Natl Acad Sci U S A. 1998;
95:2547–2551.
57. Tamura N, Ogawa Y, Chusho H, Nakamura K, Nakao K, Suda M,
Kasahara M, Hashimoto R, Katsuura G, Mukoyama M, Itoh H, Saito Y,
Tanaka I, Otani H, Katsuki M. Cardiac fibrosis in mice lacking brain
natriuretic peptide. Proc Natl Acad Sci U S A. 2000;97:4239 – 4244.
58. Van der Zander K, Houben AJ, Kroon AA, de Leeuw PW. Effects of
brain natriuretic peptide on forearm vasculature: comparison with atrial
natriuretic peptide. Cardiovasc Res. 1999;44:595– 600.
59. Goy MF, Oliver PM, Purdy KE, Knowles JW, Fox JE, Mohler PJ, Qian
X, Smithies O, Maeda N. Evidence for a novel natriuretic peptide
receptor that prefers brain natriuretic peptide over atrial natriuretic
peptide. Biochem J. 2001;358:379 –387.
60. Brenner BM, Ballermann BJ, Gunning ME, Zeidel ML. Diverse biological actions of atrial natriuretic peptide. Physiol Rev. 1990;70:
665– 699.
61. Johnston CI, Hodsman PG, Kohzuki M, Casley DJ, Fabris B, Phillips
PA. Interaction between atrial natriuretic peptide and the renin angiotensin aldosterone system: endogenous antagonists. Am J Med. 1989;
87:24S–28S.
62. Kudo T, Baird A. Inhibition of aldosterone production in the adrenal
glomerulosa by atrial natriuretic factor. Nature. 1984;312:756 –757.
63. Marin-Grez M, Fleming JT, Steinhausen M. Atrial natriuretic peptide
causes pre-glomerular vasodilatation and post-glomerular vasoconstriction in rat kidney. Nature. 1986;324:473– 476.
64. Cogan MG. ANF can increase renal solute excretion primarily by raising
glomerular filtration. Am J Physiol. 1986;250:F710 –F714.
65. Kiberd BA, Larson TS, Robertson CR, Jamison RL. Effect of atrial
natriuretic peptide on vasa recta blood flow in the rat. Am J Physiol.
1987;252:F1112–F1117.
66. Sonnenberg H, Honrath U, Chong CK, Wilson DR. Atrial natriuretic
factor inhibits sodium transport in medullary collecting duct. Am J
Physiol. 1986;250:F963–F966.
67. Sultanian R, Deng Y, Kaufman S. Atrial natriuretic factor increases
splenic microvascular pressure and fluid extravasation in the rat.
J Physiol. 2001;533:273–280.
68. Winquist RJ, Faison EP, Waldman SA, Schwartz K, Murad F, Rapoport
RM. Atrial natriuretic peptide elicits an endothelium-independent
relaxation and activates particulate guanylate cyclase in vascular smooth
muscle. Proc Natl Acad Sci U S A. 1984;81:7661–7664.
69. Holschermann H, Noll T, Hempel A, Piper HM. Dual role of cGMP in
modulation of macromolecule permeability of aortic endothelial cells.
Am J Physiol. 1997;272:H91–H98.
70. Melo LG, Veress AT, Ackermann U, Steinhelper ME, Pang SC, Tse Y,
Sonnenberg H. Chronic regulation of arterial blood pressure in ANP
transgenic and knockout mice: role of cardiovascular sympathetic tone.
Cardiovasc Res. 1999;43:437– 444.
71. Imaizumi T, Takeshita A, Higashi H, Nakamura M. ␣-ANP alters reflex
control of lumbar and renal sympathetic nerve activity and heart rate.
Am J Physiol. 1987;252:R878 –R882.
72. Nonoguchi H, Sands JM, Knepper MA. Atrial natriuretic factor inhibits
vasopressin-stimulated osmotic water permeability in rat inner medullary collecting duct. J Clin Invest. 1988;82:1383–1390.
73. Osol G, Halpern W, Tesfamariam B, Nakayama K, Weinberg D. Synthetic atrial natriuretic factor does not dilate resistance-sized arteries.
Hypertension. 1986;8:606 – 610.
74. Holtwick R, Gotthardt M, Skryabin B, Steinmetz M, Potthast R, Zetsche
B, Hammer B, Herz J, Kuhn M. Smooth muscle-selective deletion of
guanylyl cyclase-A prevents the acute but not chronic effects of ANP on
blood pressure. Proc Natl Acad Sci U S A. 2002;99:7142–7147.
75. Gardner DG, Cao L. Natriuretic peptides inhibit DNA synthesis in
cardiac fibroblasts. Hypertension. 1995;25:227–234.
76. Calderone A, Thaik CM, Takahashi N, Chang DL, Colucci WS. Nitric
oxide, atrial natriuretic peptide, and cyclic GMP inhibit the growthpromoting effects of norepinephrine in cardiac myocytes and fibroblasts.
J Clin Invest. 1998;101:812– 818.
77. Kishimoto I, Rossi K, Garbers DL. A genetic model provides evidence
that the receptor for atrial natriuretic peptide (guanylyl cyclase-A)
inhibits cardiac ventricular myocyte hypertrophy. Proc Natl Acad Sci
U S A. 2001;98:2703–2706.
78. Kuhn M, Holtwick R, Baba HA, Perriard J-C, Schmitz W, Ehler E.
Progressive cardiac hypertrophy and dysfunction in atrial natriuretic
peptide receptor (GC-A) deficient mice. Heart. 2002;87:368 –374.
79. Holtwick R, Van Eickels M, Skryabin BV, Baba HA, Bubikat A,
Begrow F, Schneider MD, Garbers DL, Kuhn M. Pressure-independent
cardiac hypertrophy in mice with cardiomyocyte-restricted inactivation
of the atrial natriuretic peptide receptor guanylyl cyclase-A. J Clin
Invest. 2003;111:1399 –1407.
80. Maack T. Receptors of atrial natriuretic factor. Annu Rev Physiol.
1992;54:11–27.
81. Murthy KS, Teng BQ, Zhou H, Jin JG, Grider JR, Makhlouf GM.
Gi-1/Gi-2-dependent signaling by single-transmembrane natriuretic
peptide clearance receptor. Am J Physiol. 2000;278:G974 –G980.
82. Matsukawa N, Grzesik WJ, Takahashi N, Pandey KN, Pang S,
Yamauchi M, Smithies O. The natriuretic peptide clearance receptor
locally modulates the physiological effects of natriuretic peptide system.
Proc Natl Acad Sci U S A. 1999;96:7403–7408.
83. Dessi-Fulgheri P, Sarzani R, Tamburrini P, Moraca A, Espinosa E, Cola
G, Giantomassi L, Rappelli A. Plasma atrial natriuretic peptide and
natriuretic peptide receptor gene expression in adipose tissue of normotensive and hypertensive obese patients. J Hypertens. 1997;15:
1695–1699.
84. Andreassi MG, Del Ry S, Palmieri C, Clerico A, Biagini A, Giannessi
D. Up-regulation of “clearance” receptors in patients with chronic heart
failure: a possible explanation for the resistance to biological effects of
cardiac natriuretic hormones. Eur J Heart Fail. 2001;3:407– 414.
85. Katafuchi T, Mizuno T, Hagiwara H, Itakura M, Ito T, Hirose S.
Modulation by NaCl of atrial natriuretic peptide receptor levels and
cyclic GMP responsiveness to atrial natriuretic peptide of cultured
vascular endothelial cells. J Biol Chem. 1992;267:7624 –7629.
86. Potter LR, Garbers DL. Dephosphorylation of the guanylyl cyclase-A
receptor causes desensitization. J Biol Chem. 1992;267:14531–14534.
87. Potter LR, Hunter T. Phosphorylation of the kinase homology domain is
essential for activation of the A-type natriuretic peptide receptor. Mol
Cell Biol. 1998;18:2164 –2172.
88. Foster DC, Garbers DL. Dual role for adenine nucleotides in the regulation of the atrial natriuretic peptide receptor, guanylyl cyclase-A.
J Biol Chem. 1998;273:16311–16318.
89. Foster DC, Wedel BJ, Robinson SW, Garbers DL. Mechanisms of
regulation and functions of guanylyl cyclases. Rev Physiol Biochem
Pharmacol. 1999;135:1–39.
90. Potter LR, Hunter T. Guanylyl cyclase-linked natriuretic peptide
receptors: structure and regulation. J Biol Chem. 2001;276:6057– 6060.
91. Jewett JR, Koller KJ, Goeddel DV, Lowe DG. Hormonal induction of
low affinity receptor guanylyl cyclase. EMBO J. 1993;12:769 –777.
92. Airhart N, Yang Y-F, Roberts CT, Silberbach M. Atrial natriuretic
peptide induces natriuretic peptide receptor-cGMP-dependent protein
kinase interaction. J Biol Chem. 2003;278:38693–38698.
93. Potter LR, Garbers DL. Protein kinase C-dependent desensitization of
the atrial natriuretic peptide receptor is mediated by dephosphorylation.
J Biol Chem. 1994;269:14636 –14642.
94. Haneda M, Kikkawa R, Maeda S, Togawa M, Koya D, Horide N,
Kajiwara N, Shigeta Y. Dual mechanism of angiotensin II inhibits
ANP-induced mesangial cGMP accumulation. Kidney Int. 1991;40:
188 –194.
95. Yasunari K, Kohno M, Murakawa K, Yokokawa K, Horio T, Takeda T.
Phorbol ester and atrial natriuretic peptide receptor response on vascular
smooth muscle. Hypertension. 1992;19:314 –319.
96. Christman TD, Garbers DL. Reciprocal antagonism coordinates C-type
natriuretic peptide and mitogen-signaling pathways in fibroblasts. J Biol
Chem. 1999;274:4293– 4299.
Kuhn
Downloaded from http://circres.ahajournals.org/ by guest on July 31, 2017
97. Pandey KN, Nguyen HT, Sharma GD, Shi S-J, Kriegel AM. Ligandregulated internalization, trafficking, and down-regulation of guanylyl
cyclase/atrial natriuretic peptide receptor-A in human embryonic kidney
293 cells. J Biol Chem. 2002;277:4618 – 4627.
98. Iimura O, Kusano E, Ishida F, Oono S, Ando Y, Asano Y. Hyperosmolality rapidly reduces atrial-natriuretic-peptide-dependent cyclic
guanosine monophosphate production in cultured rat inner medullary
collecting duct cells. Pflugers Arch. 1995;430:81– 87.
99. Shinohara M, Nonoguchi H, Ujiie K, Terada Y, Owada A, Tonita K,
Marumo F. Effects of hyperosmolality on ANP-stimulated cGMP generation in rat inner medullary collecting duct. Kidney Int. 1994;45:
1362–1368.
100. Berl T, Mansour J, Veis JH. Regulation of atrial natriuretic peptidestimulated cGMP production in the inner medulla. Kidney Int. 1992;41:
37– 42.
101. Chen S, Gardner DG. Osmoregulation of natriuretic peptide receptor
signaling in inner medullary collecting duct: a requirement for p38
MAPK. J Biol Chem. 2002;277:6037– 6043.
102. Cao L, Wu J, Gardner DG. Atrial natriuretic peptide suppresses the
transcription of its guanylyl cyclase-linked receptor. J Biol Chem. 1995;
270:24891–24897.
103. Roubert P, Lonchampt MO, Chabrier PE, Plas P, Goulin J, Braquet P.
Down-regulation of atrial natriuretic factor receptors and correlation
with cGMP stimulation in rat cultured vascular smooth muscle cells.
Biochem Biophys Res Commun. 1987;148:61– 67.
104. Tremblay J, Desjardins R, Hum D, Gutkowska J, Hamet P. Biochemistry
and physiology of the natriuretic peptide receptor guanylyl cyclases. Mol
Cell Biochem. 2002;230:31– 47.
105. Zhang LM, Tao H, Newman WH. Regulation of atrial natriuretic peptide
receptors in vascular smooth muscle cells: role of cGMP. Am J Physiol.
1993;264:H1753–H1759.
106. Garg R, Pandey KN. Angiotensin II–mediated negative regulation of Npr1
promoter activity and gene transcription. Hypertension. 2003;41:730–736.
107. Ye Q, Chen S, Gardner DG. Endothelin inhibits NPR-A and stimulates
eNOS gene expression in rat IMCD cells. Hypertension. 2003;41:
675– 681.
108. Rutledge DR, Sun Y, Ross EA. Polymorphisms within the atrial natriuretic peptide gene in essential hypertension. J Hypertens. 1995;13:
953–955.
109. Beige J, Ringel J, Hohenbleicher H, Rubattu S, Kreutz R, Sharma AM.
HpaII-polymorphism of the atrial-natriuretic-peptide gene and essential
hypertension in whites. Am J Hypertens. 1997;10:1316 –1318.
110. Nakayama T, Soma M, Takahashi Y, Rehemudula D, Kanmatsuse K,
Furuya K. Functional deletion mutation of the 5⬘-flanking region of type
A human natriuretic peptide receptor gene and its association with
essential hypertension and left ventricular hypertrophy in the Japanese.
Circ Res. 2000;86:841– 845.
111. Bulut D, Potthast R, Hanefeld C, Schulz T, Kuhn M, Mügge A. Impaired
vasodilator responses to atrial natriuretic peptide in essential hypertension. Eur J Clin Invest. 2003;33:567–573.
Membrane Guanylyl Cyclase Receptors
709
112. Knowles JW, Erickson LM, Guy VK, Sigel CS, Wilder JC, Maeda N.
Common variations in noncoding regions of the human natriuretic
peptide receptor A gene have quantitative effects. Hum Genet. 2003;
112:62–70.
113. Sarzani R, Dessi-Fulgheri P, Salvi F, Serenelli M, Spagnolo D, Cola G,
Pupita M, Giantomassi L, Rappelli A. A novel promoter variant of the
natriuretic peptide clearance receptor gene is associated with lower atrial
natriuretic peptide and higher blood pressure in obese hypertensives.
Hypertension. 1999;17:1301–1305.
114. Campese VM, Tawadrous M, Bigazzi R, Bianchi S, Mann AS, Oparil S,
Raij L. Salt intake and plasma atrial natriuretic peptide and nitric oxide
in hypertension. Hypertension. 1996;28:335–340.
115. Sala C, Ambrosi B, Morganti A. Blunted vascular and renal effects of
exogenous atrial natriuretic peptide in patients with Cushing’s disease.
J Clin Endocrinol Metab. 2001;86:1957–1961.
116. Burnett JC Jr, Kao PC, Hu DC, Heser DW, Heublein D, Granger JP,
Opgenorth TJ, Reeder GS. Atrial natriuretic peptide in congestive heart
failure in the human. Science. 1986;231:1145–1147.
117. Hirooka Y, Takeshita A, Imaizumi T, Suzuki S, Yoshida M, Ando S,
Nakamura M. Attenuated forearm vasodilative response to intra-arterial
atrial natriuretic peptide in patients with heart failure. Circulation. 1990;
82:147–153.
118. Schiffrin EL. Decreased density of binding sites for atrial natriuretic
peptide on platelets of patients with severe congestive heart failure. Clin
Sci (Lond). 1988;74:213–218.
119. Kook H, Itoh H, Choi BS, Sawada N, Doi K, Hwang TJ, Kim KK, Arai
H, Baik YH, Nakao K. Physiological concentration of atrial natriuretic
peptide induces endothelial regeneration in vitro. Am J Physiol. 2003;
284:H1388 –H1397.
120. Carini R, De Cesaris MG, Splendore R, Domenicotti C, Nitti MP,
Pronzato MA, Albano E. Mechanisms of hepatocyte protection against
hypoxic injury by atrial natriuretic peptide. Hepatology. 2003;37:
277–285.
121. Sengenés C, Berlan M, De Glisezinski I, Lafontan M, Galitzky J.
Natriuretic peptides: a new lipolytic pathway in human adipocytes.
FASEB J. 2000;14:1345–1351.
122. Zolle O, Lawrie AM, Simpson AW. Activation of the particulate and not
the soluble guanylate cyclase leads to the inhibition of Ca2⫹ extrusion
through localized elevation of cGMP. J Biol Chem. 2000;275:
25892–25899.
123. Colucci WS, Elkayam U, Horton DP, Abraham WT, Bourge RC,
Johnson AD, Wagoner LE, Givertz MM, Liang CS, Neibaur M, Haught
WH, LeJemtel TH. Intravenous nesiritide, a natriuretic peptide, in the
treatment of decompensated congestive heart failure: Nesiritide Study
Group. N Engl J Med. 2000;343:246 –253.
124. Yu S, Avery L, Baude E, Garbers DL. Guanylyl cyclase expression in
specific sensory neurons: a new family of chemosensory receptors. Proc
Natl Acad Sci U S A. 1997;94:3384 –3387.
Structure, Regulation, and Function of Mammalian Membrane Guanylyl Cyclase
Receptors, With a Focus on Guanylyl Cyclase-A
Michaela Kuhn
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Circ Res. 2003;93:700-709
doi: 10.1161/01.RES.0000094745.28948.4D
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