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ANNUAL
REVIEWS
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Structure and Regulation of
Soluble Guanylate Cyclase
Emily R. Derbyshire1 and Michael A. Marletta2
1
Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical
School, Boston, Massachusetts 02115
2
Department of Chemistry, The Scripps Research Institute, La Jolla, California 92037;
email: [email protected]
Annu. Rev. Biochem. 2012. 81:533–59
Keywords
First published online as a Review in Advance on
February 9, 2012
nitric oxide, heme, signaling, desensitization, nitrosation
The Annual Review of Biochemistry is online at
biochem.annualreviews.org
Abstract
This article’s doi:
10.1146/annurev-biochem-050410-100030
c 2012 by Annual Reviews.
Copyright All rights reserved
0066-4154/12/0707-0533$20.00
Nitric oxide (NO) is an essential signaling molecule in biological systems. In mammals, the diatomic gas is critical to the cyclic guanosine monophosphate (cGMP) pathway as it functions as the primary
activator of soluble guanylate cyclase (sGC). NO is synthesized from
L-arginine and oxygen (O2 ) by the enzyme nitric oxide synthase (NOS).
Once produced, NO rapidly diffuses across cell membranes and binds
to the heme cofactor of sGC. sGC forms a stable complex with NO
and carbon monoxide (CO), but not with O2 . The binding of NO to
sGC leads to significant increases in cGMP levels. The second messenger then directly modulates phosphodiesterases (PDEs), ion-gated
channels, or cGMP-dependent protein kinases to regulate physiological
functions, including vasodilation, platelet aggregation, and neurotransmission. Many studies are focused on elucidating the molecular mechanism of sGC activation and deactivation with a goal of therapeutic
intervention in diseases involving the NO/cGMP-signaling pathway.
This review summarizes the current understanding of sGC structure
and regulation as well as recent developments in NO signaling.
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INTRODUCTION
Contents
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INTRODUCTION . . . . . . . . . . . . . . . . . .
Enzymes Critical to the Nitric
Oxide/Cyclic Guanosine
Monophosphate Pathway . . . . . . . .
ISOLATION OF SOLUBLE
GUANYLATE CYCLASE . . . . . . . .
SOLUBLE GUANYLATE
CYCLASE ISOFORMS . . . . . . . . . . .
ARCHITECTURE OF SOLUBLE
GUANYLATE CYCLASE . . . . . . . .
Heme-Nitric Oxide and Oxygen
Binding Domain . . . . . . . . . . . . . . . .
Per/Arnt/Sim and Coiled-Coil
Domains . . . . . . . . . . . . . . . . . . . . . . .
Catalytic Domain . . . . . . . . . . . . . . . . . .
SOLUBLE GUANYLATE
CYCLASE HOMOLOGS . . . . . . . . .
Eukaryotic Atypical Soluble
Guanylate Cyclases . . . . . . . . . . . . .
Prokaryotic Heme-Nitric Oxide
and Oxygen Binding
Proteins . . . . . . . . . . . . . . . . . . . . . . . .
STRUCTURAL INSIGHTS FROM
STUDIES ON SOLUBLE
GUANYLATE CYCLASE AND
ITS HOMOLOGS . . . . . . . . . . . . . . . .
LIGAND SELECTIVITY . . . . . . . . . . . .
REGULATION OF SOLUBLE
GUANYLATE CYCLASE BY
NITRIC OXIDE . . . . . . . . . . . . . . . . . .
Activation and Nitric Oxide
Association . . . . . . . . . . . . . . . . . . . . .
Deactivation and Nitric Oxide
Dissociation . . . . . . . . . . . . . . . . . . . .
Desensitization . . . . . . . . . . . . . . . . . . . .
MODULATORS OF SOLUBLE
GUANYLATE CYCLASE
ACTIVITY . . . . . . . . . . . . . . . . . . . . . . .
Soluble Guanylate Cyclase
Activators . . . . . . . . . . . . . . . . . . . . . .
Soluble Guanylate Cyclase
Inhibitors. . . . . . . . . . . . . . . . . . . . . . .
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The nitric oxide/cyclic guanosine monophosphate (NO/cGMP) pathway was discovered
in the 1980s, but chemical modulation of the
pathway for the treatment of angina pectoris
had been unknowingly achieved 100 years
earlier. This stimulation of cGMP production
occurred with the clinical administration of
organic nitrites (isoamyl nitrite) (1) and organic
nitrates (glyceryl trinitrate; GTN) (2). These
compounds alleviate the pain associated with
angina by relaxing the vascular smooth muscle,
leading to vasodilation. For years, investigations were focused on the mechanism of smooth
muscle relaxation by these molecules, and these
efforts led to the discovery that NO is a physiologically relevant signaling molecule. Additionally, these efforts led to the identification of the
enzymes that biosynthesize NO and cGMP.
Enzymes Critical to the Nitric
Oxide/Cyclic Guanosine
Monophosphate Pathway
Early studies showed that both cytosolic and
particulate fractions of mammalian tissue
exhibit guanylate cyclase activity. Within
the insoluble fractions, membrane-bound
particulate guanylate cyclases are present,
which are activated by natriuretic peptides
(reviewed in References 3 and 4), whereas the
cytosolic fractions contain soluble guanylate
cyclases (sGCs), which are activated by NO.
NO-responsive guanylate cyclase activity is
also associated with cell membranes in certain
tissues, including skeletal muscle and brain, as
well as in platelets (5–7). Guanylate cyclases
are found in most tissues, and the distribution
of these proteins in various cells is isoform
specific. This provides an additional means to
regulate cGMP-dependent responses because
localized pools of the signaling molecule can
be generated within specific tissues and in
proximity to either soluble or membranebound cGMP receptors. Thus, tissues can
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regulate cGMP levels by expression of specific
GC isoforms, and the isoforms have a distinct
peptide receptor or ligand activator. Additionally, a reciprocal communication between
particulate guanylate cyclase and sGC has been
observed in the regulation of human and mouse
vascular homeostasis (8), and it remains likely
that communication between these pathways
occurs in several cGMP-dependent processes.
Generally, in eukaryotic NO signaling,
the initial event involves calcium release,
followed by binding of a calcium/calmodulin
complex to nitric oxide synthase (NOS), which
activates the enzyme. NO is synthesized and
then diffuses into target cells, where it binds
to the heme in sGC (Figure 1). sGC is a
histidine-ligated hemoprotein that binds NO
and carbon monoxide (CO), but not oxygen
(O2 ). This binding event leads to a several
hundredfold increase in cGMP synthesis. Once formed, cGMP targets include
Generator cell
phosphodiesterases (PDEs), ion-gated channels, and cGMP-dependent protein kinases
in the regulation of several physiological
functions, including vasodilation, platelet
aggregation, and neurotransmission (9–11).
In 1998, the Nobel Prize in Physiology or
Medicine was awarded to Robert F. Furchgott,
Louis J. Ignarro, and Ferid Murad, in recognition of their achievements toward the discovery
of the NO-signaling pathway. Currently, this
pathway is actively studied because drugs modulating NO-dependent processes have the potential to treat several maladies, including cardiovascular and neurodegenerative diseases, as
well as various airway diseases.
cGMP: cyclic
guanosine
monophosphate
GTN: glyceryl
trinitrate
Vasodilation: blood
vessel widening from
smooth muscle
relaxation
sGC: soluble
guanylate cyclase
NOS: nitric oxide
synthase
ISOLATION OF SOLUBLE
GUANYLATE CYCLASE
Despite many years of research on sGC, an efficient low-cost purification of the protein has
Ta
Target cell
α1 β1
Ca2+/CaM
GTP
NO
L-Arg
+ O2
FeII
Mg2+
cGMP + PPi
sGC
NOS
L-Cit
+
NO
cGK
PDE
cGMP-gated
ion channels
Figure 1
The nitric oxide/cyclic GMP (NO/cGMP)-signaling pathway. A Ca2+ /calmodulin (CaM) complex binds
nitric oxide synthase (NOS). NOS catalyzes the oxidation of L-arginine (L-Arg) to L-citrulline (L-Cit) and
nitric oxide (NO). NO binds to the FeII heme of α1β1 soluble guanylate cyclase (sGC) at a diffusioncontrolled rate. This binding event leads to significant increases in cGMP and pyrophosphate (PPi ). cGMP
then binds to and activates cGMP-dependent protein kinases (cGKs), phosphodiesterases (PDEs) and
ion-gated channels. Abbreviations: α1 and β1, soluble guanylate cyclase subunits; CaM, calmodulin.
www.annualreviews.org • Structure and Regulation of SGC
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remained elusive, but several methods have
been developed that yield low microgram
amounts of homogeneous protein. Initial characterization of sGC was carried out with protein obtained from rat and bovine tissues. By
the 1980s, studies were being done with purified sGC from rat lung (12) and liver (13),
as well as from bovine lung (14, 15); these
studies showed sGC to be a heterodimer. Importantly, it was observed that sGC could be
purified with and without the heme cofactor, depending on the purification protocol.
In short, the use of solubilizing agents or ammonium sulfate precipitation can lead to misfolded apoprotein. Heme reconstitution of this
apoprotein yields an sGC species [later termed
sGC1 by Vogel et al. (16)] with biochemical properties that vary from the native protein [named sGC2 by Vogel et al. (16)]. To
date, the bovine lung sGC prep is the most efficient method of isolating heme-bound protein from source tissue (17, 18). This method
typically yields ∼1 mg of pure protein per
kilogram of lung.
The development of heterologous expression systems for recombinant sGC expression
led to significant advances. The first successful
heterologous expression system for sGC was
accomplished in COS-7 cells (19). Although
COS-7 cells do not produce enough sGC for
protein purification, the procedure was pivotal
to establishing that sGC is an obligate heterodimer composed of α1 and β1 subunits (20).
Additionally, COS-7 cells have been used to
examine mutants and truncations of sGC via
lysate activity assays (21, 22).
The overexpression of rat sGC in insect cells
with the Sf9/baculovirus expression system was
the first procedure used to isolate pure recombinant protein (21, 23, 24). sGC expression in
insect cells was initially accomplished without
an affinity tag (21), but current protocols
involve a His tag to facilitate the purification
process (25–27). Although highly expressed,
most (>90%) of the protein is insoluble. This
method typically yields 0.2–0.4 mg of pure soluble protein per liter of culture and is now commonly used to obtain purified rat and human
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GTP: guanosine
5 -triphosphate
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sGC. A clear advantage of the Sf9/baculovirus
expression system is that it enables in vitro
characterization, including the generation
of site-directed mutants. However, both the
COS-7 and Sf9/baculovirus expression systems
facilitated the biochemical characterization of
subunit dimerization, allowed for the generation of site-directed mutants, and provided
larger quantities of enzyme for in vitro studies.
The full-length mammalian α1β1 heterodimer has not yet been isolated from a bacterial expression system, but several truncations
of sGC have been successfully obtained via expression in Escherichia coli. These proteins include N-terminal truncations of α1, β1, and
β2 (28–30), C-terminal truncations of α1 and
β1 (31, 32), and a domain within the central region of β1 (Figure 2) (31). These constructs
purify with yields ranging from 0.5 to 5 mg
a
Domain architecture of sGC
α
Heme
β
H-NOX PAS
b
CC
CAT
Characterized sGC truncations
Heme binding
β1(1–385)
β1(1–194)
β2(1–217)
GTP binding
α1(467–690)
β1(414–619)
Figure 2
Domain architecture of soluble guanylate cyclase
(sGC). (a) Heme-nitric oxide/oxygen binding
(H-NOX, yellow), Per/Arnt/Sim domain (PAS,
gray), coiled-coil domains (CC, white), and catalytic
domains (CAT, blue) are shown. (b) Characterized
heme-binding and GTP-binding truncations of rat
sGC. Heme is represented by the red parallelogram.
α1, β1, and β2 isoforms are shown.
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of pure protein per liter of culture. Recently
an E. coli expression system was used for the
full-length Manduca sexta α1β1 heterodimer
(33). This method provides low amounts (0.5–
1 mg/liter) of partially pure full-length protein,
but higher yields (1–2 mg/liter) of pure protein
were obtained by truncating the C terminus of
the α1 and β1 subunits (33). The resulting heterodimeric proteins (msGC-NT1 and msGCNT2) lack the ability to cyclize GTP but can be
purified to homogeneity. Perhaps future work
optimizing the expression conditions and/or
purification of sGC from E. coli will overcome
the current limitations in protein yield.
SOLUBLE GUANYLATE
CYCLASE ISOFORMS
Heterodimeric sGC consists of two homologous subunits, α and β. The most commonly
studied isoform is the α1β1 protein; however,
α2 and β2 subunits have also been identified
(34, 35). These proteins were first characterized in mammals, but they also exist in insects,
such as Drosophila melanogaster and M. sexta, and
in fish. Isoforms of α-subunits are highly homologous with ∼48% sequence identity, and
β-subunits have an overall sequence identity of
∼41%.
The localization of each subunit has been
studied in mammals, including humans, rats,
and cows. Both α1 and β1 subunits are expressed in most tissues, and it is well accepted
that these proteins form a physiologically
relevant heterodimer (36). By quantitative
polymerase chain reaction analysis and Western blotting, the α2 subunit is found in fewer
tissues when compared to the α1 and β1 isoforms but is highly expressed in the brain, lung,
colon, heart, spleen, uterus, and placenta (36).
Studies with purified protein have shown that
the α2β1 heterodimer exhibits ligand-binding
characteristics identical to the α1β1 heterodimer (37, 38), but a splice variant of the α2
subunit (α2i) forms a dimer with the β1 subunit
to form an inactive complex. α2i contains an
in-frame insertion of 31 amino acids within
the catalytic domain and appears to function as
a dominant-negative protein (39). Despite the
similar biochemical properties of the two physiologically relevant sGC heterodimers, they
have unique roles in cGMP signaling that may
be attributed to their varying cellular localization. Specifically, α2β1 has been associated
with the membrane in several tissues (5–7),
and consequently, α2β1 responds differently
than cytosolic α1β1 (37). In rat brain, it was
found that this association is mediated by an
interaction between the C terminus of the α2
protein and PSD-95 (postsynaptic density-95)
protein (6). Most recently, a distinct presynaptic role has been identified for α1β1 in
glutamate release in the hippocampus (40).
The β2 isoform is not ubiquitously expressed like the β1 isoform, and analysis of
mRNA levels indicates that it is found primarily
in the kidney (35). Unlike β1, the C terminus
of β2 contains a possible isoprenylation site,
but the subcellular localization of this protein
is unknown. The β2 isoform has not yet been
purified and characterized, but the protein has
been studied after transient expression in insect
cells. Using this approach, it was found that β2
does not exhibit cyclase activity when expressed
with α1 or α2 but that β2 is active in the absence
of an α-subunit, suggesting that the β2 protein
can function as a homodimer ex vivo (41). This
is in contrast to the β1 protein, which has been
isolated as an inactive homodimer after overexpression in insect cells (26). In rat kidney, the
mRNA levels of the β2 subunit were shown to
be developmentally regulated (42); however, it
remains unclear what the physiological role of
the β2 isoform is in cGMP signaling.
Recently, several genetic studies in mouse
models have emphasized the importance of
the various sGC isoforms for physiological
processes (reviewed in Reference 43). Knockout mice lacking the sGC β1 subunit exhibit
elevated blood pressure, reduced heart rate,
and dysfunction in gastrointestinal contractility
(44). Additionally, deletion of the β1 subunit
within only smooth muscle cells implicates the
loss of the protein in these cells as the cause
of hypertension in the knockout mice (45).
Deletion of the β1 subunit is generally viewed
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PAS: protein fold
named for its
association with the
Per, ARNT, and Sim
proteins
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H-NOX: heme-nitric
oxide and oxygen
binding
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as a global sGC knockout because α1 and α2
do not form functional heterodimers with β2.
sGC α1 and α2 knockout mice have also been
generated. Both proteins were found to be
essential for long-term potentiation (46), and
vasodilation was mediated primarily by the α1
isoform (47). In addition to clarifying the role of
the α1β1 protein in NO-mediated pulmonary
vasodilation (48, 49), studies with α1 subunitdeficient mice suggest that both α-subunits are
involved in gastric nitrergic relaxation (50) and
relaxation of colon tissue (51).
Several invertebrates also contain genes
that encode predicted NO-sensitive guanylate
cyclases. Likely owing to the genetic tools
available in D. melanogaster, this organism
contains the best-characterized insect guanylate cyclase. In D. melanogaster, Gycα-99B
and Gycβ-100B are orthologs of the α1 and
β1 subunits, respectively. Drosophila mutants
deficient in the production of these proteins
suggest that the NO/cGMP pathway mediates
a behavioral phenotype (52) and development
of the visual system (53), and this pathway is
important for larval foraging locomotion (54).
In other organisms, several biochemical studies
have been aimed at elucidating the significance
of the NO/cGMP-signaling pathway. As mentioned above, the full-length M. sexta α1β1
heterodimer has recently been characterized
(33). There is evidence that sGC in M. sexta is
involved in neuronal excitability (55) and odor
responsiveness (56). In mollusks, such as the
Limax marginatus and Limax maximus slugs,
sGC may modulate the electrical oscillation of
interneurons in the central olfactory pathway
(57, 58).
ARCHITECTURE OF SOLUBLE
GUANYLATE CYCLASE
The rat sGC α1 and β1 subunits are 690
and 619 amino acids in length, respectively.
These proteins are part of a large family of sGC
subunits that are conserved in eukaryotes. Generally, there is the highest sequence variability
at the N terminus of α-subunits and the greatest sequence identity at the C terminus of both
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the α- and β-proteins. Each sGC subunit consists of four distinct domains. The β1 subunit
contains a N-terminal heme-binding domain,
a Per/Arnt/Sim (PAS) domain, a coiled-coil
domain, and a C-terminal catalytic domain
(Figure 2) (reviewed in Reference 59).
Heme-Nitric Oxide and Oxygen
Binding Domain
Experiments with sGC truncations and sitedirected mutants were necessary to localize the
minimal heme-binding domain of sGC. These
experiments involved the systematic mutation
of conserved histidines (60), expression of various truncations in E. coli (29), and deletion
of the β1 N terminus (61). Taken together,
these studies showed that the β1 N terminus
constituted the heme-binding domain and suggested that histidine 105 (rat) was the proximal
heme ligand. The sGC heme-binding domain
has been localized to residues 1 to ∼194 on
the β1 subunit (28). Like the full-length sGC,
the isolated heme domain binds NO and CO,
but not O2 . The N terminus of the α1 subunit
has homology to the β1 N terminus and was
shown to have affinity for heme despite lacking
the proximal histidine ligand (30). The sGC N
terminus is part of a conserved family of proteins found in both prokaryotes and eukaryotes
(62). This family of proteins has been termed
heme-nitric oxide binding (62), sensor of nitric
oxide (63), and heme-nitric oxide and oxygen
binding (H-NOX) (64). H-NOX is the most
used abbreviation and will be used throughout this review. To date, all of the characterized H-NOX proteins bind heme as well as the
gaseous heme ligands NO and CO (reviewed in
Reference 65). Some H-NOX proteins, including β1 and β2, discriminate against O2 binding, whereas others form a stable complex with
O2 . In eukaryotes, H-NOX proteins have only
been found with the known sGC domain architecture. In bacteria, H-NOX domains can
be found as proteins of ∼200 amino acids in
length with a single predicted function or as a
domain within a larger protein on the basis of
sequence analysis programs. Additionally, the
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genes that encode these H-NOX proteins in
bacteria are in proximity to genes that encode
putative histidine kinases and diguanylate cyclases, or in some cases, a gene encodes the
H-NOX as a domain within a larger protein,
often fused to a methyl-accepting chemotaxis
domain. It is likely that these proteins have an
evolutionarily conserved function and serve as
gas sensors in prokaryotes and eukaryotes.
Per/Arnt/Sim and Coiled-Coil
Domains
The central region of sGC contains two
domains of unresolved function. The domain
closer to the N terminus is predicted to adopt a
PAS-like fold. Typically, PAS domains mediate
protein-protein interactions and have often
been found to bind heme, a flavin, or a nucleotide (66). The other domain, a coiled-coil
domain, appears to be unique to sGC and shares
no significant homology with any other protein
in the National Center for Biotechnology Information protein database (http://www.ncbi.
nlm.nih.gov/protein). The coiled-coil domain of the rat β1 subunit (residues 348–409)
was isolated as a tetramer and structurally
elucidated with X-ray crystallography (31).
This structural study, in addition to experiments involving site-directed mutagenesis of
residues on the α1 subunit (67), a bimolecular
fluorescence complementation assay in cells
(68), and structural analysis of homologs of the
sGC PAS domain (69), suggests that the central
regions of both sGC subunits are important
for the formation of a functional heterodimer.
Catalytic Domain
The C-terminal regions of the α1 and β1 proteins are highly homologous to the particulate
guanylate cyclase and adenylate cyclase catalytic
domains. In the 1990s, structural insights on
the sGC catalytic domains came from homology models on the basis of crystal structures of
the adenylate cyclase catalytic domains (70, 71).
These models identified key catalytic residues,
including two conserved aspartate residues on
the α1 subunit (D485 and D529, rat numbering), which are predicted to bind two Mg2+
ions (72). These residues are critical to catalysis as the associated metals likely activate both
the nucleotide 3 -hydroxyl and the α-phosphate
for the reaction, as well as stabilize the charge
on the β- and γ-phosphates on both the substrate and product. Additionally, β1 N548 is
proposed to orient the ribose ring for the reaction. Residues thought to be responsible for
base recognition include E473 and C541 on the
β1 subunit. Other residues on both α1 (R573)
and β1 (R552) are thought to interact with the
nucleotide triphosphate (72). With the identification of these critical residues, predictions
can be made about guanylate cyclase activity
using sequence analysis. This type of analysis
would correctly predict that the β2 isoform
could function as a homodimer but that β1, α1,
and α2 need a partner to be active.
The catalytic domains have now been localized to the C-terminal 467–690 and 414–619
residues of the α1 and β1 subunits, respectively
(32). These catalytic domains must form a
heterodimer for cGMP to be synthesized, and
in the full-length protein, the catalytic efficiency of the protein is dependent on the heme
ligation state of the β1 H-NOX domain. sGC
is highly selective for GTP as a substrate, but
the protein can also cyclize 2 -d-GTP, GTPγ-S, guanosine 5 -[β,γ-imido]-triphosphate
(GMP-PNP), ITP, UTP, and ATP (73–75).
The isolated α1cat β1cat heterodimer is also
selective for GTP but can synthesize cAMP
from ATP (76). Interestingly, the activity of
α1cat β1cat is inhibited by the presence of the
H-NOX domain [β1(1–194) or β1(1–385)]
(32).1,2 This shows that these domains interact
in trans and suggests that the NO mechanism
of activation involves the relief of an inhibitory
interaction between the H-NOX domain
and the catalytic domains. In support of this
1
α1cat and β1cat are the catalytic domains of the rat sGC α1
and sGC β1 subunits, respectively.
2
β1(1–194) represents residues 1–194 of the rat sGC β1
subunit, the minimum heme-binding domain.
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sGC homologs:
proteins with high
sequence homology to
mammalian sGCs
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Atypical sGCs:
distinct subset of sGCs
with reduced
sensitivity to NO
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proposal, a fluorescence (or Förster) resonance
energy transfer-based study showed that the
N terminus of both the α1 and α2 subunits
interact with the C terminus of the β1 subunit
(77). In addition to the heterodimeric rat sGC
catalytic domains, the catalytic domain from
the Chlamydomonas reinhardtii sGC (CYG12)
has been biochemically characterized (78).
This protein, as well as Synechocystis PCC6803
Cya2, a particulate guanylate cyclase (79), is
discussed in more detail below.
SOLUBLE GUANYLATE
CYCLASE HOMOLOGS
After the initial identification in mammals, it
was not until the emergence of genome sequencing that the prevalence of sGC and sGC
homologs in other organisms was fully realized.
Full-length guanylate cyclases with domain architecture similar to the α1 and β1 subunits
were found in several eukaryotic organisms. In
prokaryotes, sGC-like H-NOX domains were
found as stand-alone proteins or fused to other
functional domains (62). In some genomes, an
sGC-like PAS domain was also identified. To
date, the genes for sGC-like PAS domains are
always found near genes that encode sGC-like
H-NOX domains (62).
Eukaryotic Atypical Soluble
Guanylate Cyclases
As mentioned above, an increasing number
of eukaryotic organisms are known to contain
predicted sGCs. Some sGCs are very similar
to the well-characterized rat α1 and β1 subunits, whereas others vary significantly. Collectively, these cyclases have been termed atypical
sGCs (80), owing to their distinct activation and
dimerization properties. Some atypical sGCs
exhibit a very surprising property—the ability
to respond to O2 .
The D. melanogaster genome contains five
genes that code for sGCs. Two of these genes
code for subunits with high homology to the α1
and β1 proteins and have been shown to form
a highly NO-sensitive heterodimeric sGC
(Gycα-99B and Gycβ-100B). The other three
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genes code for subunits with greater homology
to the β2 protein (Gyc-88E, Gyc-89Da,
and Gyc-89Db) (81). Like β2, Gyc-88E can
function as a homodimer, whereas Gyc-89Da
and Gyc-89Db are only active as heterodimers
(81). Experiments with cells overexpressing
Gyc-88E, Gyc-89Da, and Gyc-89Db indicate
that cGMP synthesis in these cyclases is activated in the absence of O2 (80), and work with
purified protein confirms that the Gyc-88E
homodimer forms a stable complex with O2
(82). As expected, Gyc-88E also binds NO
and CO, but there are data that support the
role of these proteins in mediating behavioral responses in hyperoxic environments in
D. melanogaster (83). Interestingly, Gyc-88E
is inhibited two- to threefold by the binding
of NO, CO, and O2 , a property that is quite
distinct from the ligand-induced activation of
the sGC α1β1 heterodimer.
On the basis of sequence analysis, atypical sGCs exist in several organisms, including Caenorhabditis elegans (GCY-31-GCY-37);
however, worms do not contain a predicted
NOS or NO-sensitive sGC. Seven β-like
guanylate cyclases are contained within the
C. elegans genome, and it is likely that each gene
has an important functional role. GCY-35 is involved in social feeding (84), and both GCY-35
and GCY-36 promote aggregation and bordering behaviors (85). Significantly, these cGMPdependent behavioral responses are mediated
by O2 , which suggests the proteins function as
O2 sensors in vivo (84, 86). In support of this
proposal, the GCY-35 H-NOX domain was
isolated and shown to bind O2 (84). Analysis of
gcy-31 and gcy-33 mutants also implicates these
genes in O2 -dependent behavioral responses in
C. elegans, and the proteins encoded by these
genes (GCY-31 and GCY-33) likely function
in distinct sensory neurons (BAG versus URX)
when compared to GCY-35 and GCY-36 (87).
Thus, a distinct class of cyclases exist, which
bind O2 and are inhibited by ligand binding, but
there is currently no means to predict if a cyclase is activated or inhibited by gaseous ligand
binding on the basis of sequence analysis. However, residues that contribute to gaseous ligand
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selectivity have been identified, as described
below.
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Prokaryotic Heme-Nitric Oxide
and Oxygen Binding Proteins
Several species of bacteria are known to contain
H-NOX proteins, and this number continues
to increase as more genomes are sequenced. Interestingly, all of the isolated H-NOX domains
from facultative aerobes have ligand-binding
properties like sGC, namely they do not
bind O2 . In facultative aerobes, the bacterial
members of the H-NOX family encode a single
domain as a predicted stand-alone protein,
and genes that encode either putative histidine
kinases or diguanylate cyclases are found in
the same predicted operon, suggesting that the
domain has a role in two-component signaling
in bacteria. In support of this hypothesis, an
H-NOX domain and a predicted histidine
kinase from Shewanella oneidensis were isolated
and found to interact in vitro. Additionally, the
functional interaction between the H-NOX
and kinase was mediated by NO (88). Vibrio fischeri also contains an H-NOX protein, termed
H-NOXVf , which is proposed to regulate a putative histidine kinase. In V. fischeri, H-NOXVf
was shown to regulate genes involved in iron
uptake, and these same genes are modulated
by the presence of NO (89), suggesting that
H-NOXVf mediates gene expression by sensing
NO. V. fischeri colonizes the light-emitting
organ of the Hawaiian bobtail squid, Euprymna
scolopes, and it is likely that this mutualistic
host-microbe symbiosis is mediated by a
NO/H-NOXVf interaction. Some bacteria,
like Rhodobacter sphaeroides and Nostoc punctiforme, also encode a predicted PAS-like domain
upstream of the H-NOX gene (62). The functional significance of this PAS domain to
bacterial signaling is unknown, but it has been
proposed to mediate protein dimerization (69).
In obligate anaerobes, such as Thermonanaerobacter tengcongensis, the N-terminal
H-NOX domain is fused to a C-terminal
methyl-accepting chemotaxis protein, suggesting a role in a chemotactic/signaling function.
This H-NOX domain was found to form a
NITRIC OXIDE SIGNALING IN BACTERIA
A currently expanding topic within biological studies on NO involves the role of the diatomic gas in bacterial signaling. NOS-like
proteins have been identified in several prokaryotic organisms,
including Bacillus anthracis, Bacillus subtilis, Sorangium cellulosum,
and Streptomyces turgidiscabies. Additionally, a wide range of bacteria also have nitrite reductases, which generate NO as part of
denitrifying, assimilatory, and dissimilatory pathways. This endogenously produced NO is known to regulate several transcription factors via S-nitrosation. There are also two potential classes
of prokaryotic heme-based NO sensors: globin-like proteins and
H-NOX proteins. Microbial globin-like proteins bind O2 , NO,
and CO, and are thought to be involved in the nitrosative stress
response. Some H-NOX proteins bind O2 , in addition to CO
and NO, whereas others exclude O2 binding. The genes that
code for these H-NOX proteins are found in the same operons
as predicted histidine kinases or diguanylate cyclases. In vitro
studies have shown that the H-NOX protein and histidine kinase
from S. oneidensis interact in a NO-dependent manner, but the
physiological significance of this interaction is unknown. However, NO is known to be important for the symbiosis between the
bobtail squid and V. fischeri, where the H-NOX from V. fischeri
regulates colonization of the bacteria within the symbiotic host
squid by a mechanism that is dependent on NO. Further experiments in different microbial systems will likely uncover additional
NO-dependent signaling processes in bacteria.
very stable heme-O2 complex (Kd = 90 nM)
(90), a molecular distinction from the H-NOX
domains from aerobic bacteria. The isolation
and characterization of the T. tengcongensis
H-NOX domain have significantly influenced
current understanding of sGC because it was
the first H-NOX domain to be structurally
determined, and, moreover, it was crystallized
bound to the diatomic ligand O2 (63, 64).
Thus far, these proteins have been used as
tools for probing sGC structure and regulation, but functional studies in microbial systems
(see the sidebar titled Nitric Oxide Signaling
in Bacteria) will be particularly interesting as
the variable ligand-binding properties of these
H-NOXs may have consequences for their ability to respond to different gases, i.e., some proteins may sense O2 in addition to NO or CO.
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H-NOX domain
PAS domain
CC domain
Catalytic domain
Figure 3
Structures of heme nitric oxide and oxygen binding (H-NOX), PAS, coiled-coil (CC), and catalytic domains. The structures shown are
the T. tengcongensis H-NOX domain [Protein Data Bank (pdb) code 1U55], the dimerized PAS domain from N. punctiforme PCC 73102
(pdb 2p04), the CC domain from the rat β1 subunit (pdb 3hls), and the homodimeric guanylate cyclase domain from C. reinhardtii
(pdb 3et6).
STRUCTURAL INSIGHTS
FROM STUDIES ON SOLUBLE
GUANYLATE CYCLASE AND
ITS HOMOLOGS
YC-1:
3-(5 -hydroxymethyl2 -furyl)-1-benzyl
indazole
542
An increasing number of sGC homologs have
been isolated and characterized. Significantly,
some of these homologs have been amenable
to crystallography, thus providing a foundation
for structural proposals on the β1 H-NOX,
PAS, and catalytic domains. The first crystal
structure of an sGC-like domain was that of
the O2 -binding H-NOX domain (residues
1–188) from T. tengcongensis (Figure 3) (63,
64). This protein crystallized in two different
six-coordinate heme states, with O2 bound
to the reduced heme iron and in the oxidized heme state. These reports identified
several amino acids with critical structural
roles that are highly conserved within the
H-NOX family. Among the highly conserved
amino acids was the T. tengcongensis H-NOX
heme-coordinating histidine (H102) and
three residues that stabilize heme binding
(Figure 4). Specifically, arginine (R135) is critical to the coordination of the heme propionate
groups and forms a hydrogen bond with both
carboxyl groups, and tyrosine (Y135) and serine
(S133) coordinate to one of the heme carboxyl
groups. Together, these residues form a YxSxR
motif that is strictly conserved in H-NOX
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proteins. The proximal histidine, arginine,
and tyrosine had been previously identified as
critical residues for heme binding in the sGC β1
subunit on the basis of mutagenesis studies (91,
92), and the role of these residues became clear
when the T. tengcongensis H-NOX structure was
solved.
Another striking characteristic of the
T. tengcongensis H-NOX structure is a highly
nonplanar heme conformation; it contains one
of the most highly distorted hemes reported in
the Protein Data Bank. Interestingly, different
molecules of the O2 -bound H-NOX structure
exhibit varying degrees of deformation, suggesting that the heme can exist in a range of conformations. This proposal is supported by the
observation that the crystal structure of the HNOX protein from Nostoc sp. contains a moderately distorted heme (93). This heme distortion
is not an artifact of crystallization as it is also
observed in solution on the basis of an NMR
study of the S. oneidensis H-NOX (94) and resonance Raman experiments with T. tengcongensis
H-NOX (95) and α1β1 sGC (96–98). In fact,
the dynamic range of heme conformations can
be accessed in solution by site-directed mutagenesis (95) or, in the case of sGC, by addition
of the allosteric activators YC-1 or BAY 412272 (96–98). Specifically, the α1β1 sGC heme
becomes more planar upon activator binding;
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αA
αD
αG
Y140
αF
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N74
W9
H102
Figure 4
Distal heme pocket of the T. tengcongensis H-NOX domain [Protein Data Bank (pdb) 1U55 code]. Residues
important to ligand selectivity (W9, N74, and Y140) and heme binding (H102) are shown. Image
reproduced with permission of Elsevier (65).
however, it is unclear if this is a cause or consequence of enzyme activation. Thus, although
the functional importance of heme distortion
in sGC activation remains unknown, there is
a potential to utilize changes in distortion in
biological responses.
Among the residues thought to be critical to
maintaining the nonplanar heme conformation
are I5, D45, R135 (in the YxSxR motif ), L144,
and P115, all of which are highly conserved
in H-NOX proteins (64). In T. tengcongensis
H-NOX, mutation of proline 115 to alanine
leads to relaxation of the distorted heme (95,
99). This heme relaxation was not observed after mutation of proline 118 to alanine (P115 in
T. tengcongensis H-NOX) in α1β1 sGC (96);
however, it is not surprising that additional
residues or domain interactions are important
for maintaining the heme conformation in the
mammalian protein.
The crystal structure of the non-O2 -binding
H-NOX from Nostoc sp. has been solved in
the five-coordinate unligated state and as the
six-coordinate NO- and CO-heme complexes
(93). Nostoc sp. H-NOX could function as a
redox or NO sensor (100). Comparison of the
T. tengcongensis H-NOX and Nostoc sp. H-NOX
structures in different ligation states (FeII -CO,
FeII -NO, and FeII -unligated states) led to
speculation on a molecular mechanism of sGC
activation. Specifically, the differential pivoting
and bending in the H-NOX heme upon NO or
CO binding was suggested to account for the
varying degree of activation induced by the two
ligands (200-fold versus fourfold, respectively)
(93). In support of this proposal, an UV
resonance Raman study found that significant
conformational changes occurred at the N
terminus of sGC upon activator binding (96).
In sGC, the breaking of the proximal iron
(Fe)-His bond is an essential event in the
activation by NO, and thus, a structure of a
five-coordinate H-NOX protein would be very
important. Although a five-coordinate NO
complex has not yet been determined, there are
two recently solved structures where the net
effect is the severing of the proximal Fe-His
bond; the solution structure of a S. oneidensis HNOX mutant (94) and the crystal structure of
BAY 58-2667-bound Nostoc sp. H-NOX (101).
Wild-type S. oneidensis H-NOX was shown to
modulate the activity of a histidine kinase in
a ligand-dependent manner: Kinase activity
is inhibited by the FeII -NO H-NOX but not
the FeII -unligated H-NOX (88). The H103G
FeII -CO mutant mimics the kinase-inhibitory
activity of the wild-type FeII -NO H-NOX (94).
Because H103 is the proximal iron ligand in
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S. oneidensis H-NOX, mutation of this residue
to glycine leads to the isolation of apoprotein.
Similar to previous reports with other heme
proteins (92, 102), heme binding in S. oneidensis
H-NOX H103G can be rescued by imidazole.
H103 is in α-helix F, and in the heme-binding
rescued mutant, this helix is free to adopt a
position like that in a five-coordinate NO
complex. The S. oneidensis H103G H-NOX
structure, along with the structure of the
weakly active wild-type FeII -CO S. oneidensis
H-NOX, was solved by NMR. The solution
structures of both of these proteins indicate
that major changes in heme planarity and
H-NOX conformation occur upon cleavage of
the proximal histidine heme ligand.
Major structural changes are also observed
between the unbound and BAY 58-2667bound Nostoc sp. H-NOX structures (101).
BAY 58-2667 is a NO- and heme-independent
sGC activator (103) and is a candidate to treat
decompensated heart failure. The crystal structure shows that BAY 58-2667 is able to displace
the heme and occupy the heme-binding site,
thereby leading to a shift in the α-helix F that
contains the proximal histidine residue. Additional experiments mutating residues around
the proximal histidine on α-helix F suggest that
D102 could play a critical role in enzyme activation (104). In addition to providing a structural
basis for proposals on sGC activation, these
H-NOX structures enabled the development
of homology models of the heme-binding
domains of both NO-sensitive and atypical cyclases (28, 63, 105, 106), as well as density functional theory analysis and computer simulations
to address questions concerning structural
dynamics (107, 108).
The crystal structure of a domain from the
N. punctiforme signal transduction histidine kinase has also been determined (Figure 3). This
domain has high sequence identity (35%–38%)
to the sGC PAS domain, and the crystal structure showed that the domain dimerized and
adopted a PAS fold (69). There is no structure of a eukaryotic sGC PAS domain, but the
crystal structure of the coiled-coil domain of
the β1 subunit has been solved (Figure 3) (31).
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This structure indicates that the coiled-coil domain forms a tetramer composed of a dimer
of dimers. In addition to identifying potential
residues involved in mediating dimerization,
the authors propose that interhelix salt-bridge
formation selects for heterodimerization versus
homodimerization in sGCs on the basis of their
structure (31).
The catalytic domains of two different functional guanylate cyclases (78, 79), and the inactive human β1β1 catalytic domains [Protein
Data Bank (pdb) code 2WZ1], have been structurally elucidated (Figure 3). The functional
cyclase domains include a soluble homodimeric
guanylate cyclase from the eukaryotic algae
C. reinhardtii (78) and a particulate homodimeric guanylate cyclase from the unicellular
cyanobacterium Synechocystis PCC6803 Cya2
(79). Both proteins cyclize GTP and likely
contain two catalytic sites per dimer. On the
basis of kinetic analysis of cGMP synthesis, the
eukaryotic algae catalytic domains exhibit positive cooperativity with a Hill coefficient of 1.5.
This is similar to the cooperativity observed
in particulate guanylate cyclases and may be a
common feature of homodimeric sGCs (78).
The α1β1 heterodimer contains one active site
and a proposed pseudosymmetric site. This
proposed pseudosymmetric site is thought
to constitute an allosteric nucleotide-binding
site that communicates with the catalytic
nucleotide-binding site. On the basis of the
algae sGC structure, this communication may
involve residues contained on the β2-β3 loop
of each catalytic domain monomer. Specifically,
the interaction of D527 or E523 (C. reinhardtii
sGC numbering) with a nucleotide in one
active site could alter the loop conformation
and thus lead to a conformational change in
the other nucleotide-binding site (78).
Unfortunately, both structures are in an
inactive state, but they confirm the guanylate
cyclase residues critical to metal binding and
nucleotide recognition that were predicted
from the adenylate cyclase crystal structures.
Together, the reports of the C. reinhardtii
sGC and Cya2 catalytic domains provided the
first structures of a guanylate cyclase, and the
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structures show that there is high homology
between guanylate cyclase and adenylate
cyclase domains; however, the elucidation of
a mammalian heterodimeric structure remains
an important task for understanding sGC
regulation. Details about how movement in
the β1 H-NOX domain affects the catalytic
domain, and the role of the PAS and coiled-coil
domain in relaying a signal from the H-NOX
domain, may remain open questions until the
full-length sGC structure is elucidated.
LIGAND SELECTIVITY
The α1β1 heme environment is unique when
compared to the globins because the cofactor
efficiently binds NO while having no affinity
for O2 . Additionally, the α1β1 heterodimer exhibits an extremely slow rate of oxidation and
has among the highest midpoint potential reported for a high-spin heme protein (+187 mV
versus +58 mV for myoglobin) (109). This ability of mammalian sGC to select against O2
binding is important for it to function as a NO
sensor, as O2 is present at much higher levels
than NO in vivo, and FeII -O2 proteins react
rapidly with NO. Hemoproteins in the FeIII
oxidation state also form weak NO complexes
and could inadvertently serve as a sink for NO.
Since the discovery of sGC, several potential
mechanisms of ligand discrimination against
O2 have been proposed, including a weak FeHis bond strength, a negatively charged distal
pocket, and a sterically constrained distal pocket
(109–111). One proposal on the mechanism by
which α1β1 excludes O2 is based on analysis of
the T. tengcongensis H-NOX crystal structure
(63, 64). T. tengcongensis H-NOX stabilizes O2
binding with a hydrogen-bonding network involving a tyrosine (Y140), a tryptophan (W9),
and an asparagine (N74) (Figure 4). These
residues appear to be absent in the β1 H-NOX
protein and other O2 -excluding H-NOXs on
the basis of primary sequence alignments and
homology modeling. Conversely, several atypical sGCs, including the known O2 -binding cyclase Gyc-88E, encode a tyrosine that aligns
with T. tengcongensis H-NOX Y140. Therefore,
the absence of a hydrogen bond donor in the
α1β1 distal heme pocket, and the subsequent
increase in the O2 off rate, likely contributes
to the protein’s inability to form a stable FeII O2 complex (64, 90). Molecular dynamics simulations of O2 -binding and non-O2 -binding HNOX proteins suggest that they have varying
tunnel systems, which may also contribute to
their different ligand-binding properties (112),
and site-directed mutagenesis within the proposed tunnel indicates that it is important for
diffusion of gaseous ligands (113).
Several biochemical studies aimed at
probing ligand selectivity in sGC have been
reported. The distal-pocket tyrosine in the
O2 -binding T. tengcongensis H-NOX was
mutated to leucine (Y140L), and this mutation
significantly reduced O2 affinity. Additionally,
the introduction of a distal-pocket tyrosine in a
non-O2 -binding H-NOX from Legionella pneumophilia (L2 H-NOX) enabled the protein to
bind O2 (90). Mutagenesis studies introducing
a tyrosine into the distal pocket of the β1 HNOX-PAS domain β1(1–385) also produced a
protein that was capable of binding O2 (90), but
the same mutation in full-length sGC did not
facilitate O2 binding (106, 114). However, the
introduction of a tyrosine and glutamine into
the β1 heme pocket (I145Y/I149Q) resulted in
a full-length α1β1 protein with altered reactivity to O2 (105). A homology model of the O2 binding guanylate cyclase Gyc-88E places these
amino acids within the predicted distal heme
pocket. Thus, some O2 -binding guanylate cyclases may utilize a Tyr/Gln hydrogen-bonding
network, similar to several truncated globins
(115–118), to stabilize ligand binding. The
marked variability in O2 reactivity between fulllength sGC and heme-binding truncations of
the β1 protein highlights the potential for other
sGC domains to influence ligand selectivity.
Thus, despite significant progress in our understanding of ligand discrimination in sGC, including the identification of residues critical for
stabilizing O2 binding in sGC homologs, there
remain some unknown variables that may contribute to the ligand specificity of these heme
proteins.
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REGULATION OF SOLUBLE
GUANYLATE CYCLASE BY
NITRIC OXIDE
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Physiological responses to NO, such as smooth
muscle relaxation, are rapidly induced by low
levels of the diatomic gas. In cells, cGMP
levels rise within milliseconds after exposure
to nanomolar concentrations of NO, and this
fast response occurs because sGC efficiently
binds to and is activated by NO. When NO
dissociates from the protein, the amount of
cGMP decreases to a basal level. Thus, both
the rise and fall of cGMP levels must be tightly
regulated for proper function of cGMPdependent processes. Upon repeated exposure
to NO or NO-donors, like GTN, maximal
sGC activation decreases, and this process is
called sGC desensitization. sGC activation,
deactivation, and desensitization have been
extensively studied in vivo and in vitro. A major
challenge with understanding these processes
occurs when reconciling discrepancies in
results obtained with purified protein versus
sGC examined in the cellular milieu.
Activation and Nitric
Oxide Association
As mentioned above, the activation of sGC
in vitro and in vivo is rapid (occurs in milliseconds). Using a NO donor or NO gas,
the apparent 50% effective concentration for
NO acting on purified sGC is between 80 and
250 nM (119, 120). A similar potency of NO
has been measured in rat cerebellar cells (apparent 50% effective concentration ∼45 nM)
(121). On the basis of studies with purified
protein, it is known that NO binds to the heme
of sGC at a diffusion-controlled rate to form an
initial six-coordinate complex, which rapidly
converts to a five-coordinate ferrous nitrosyl
(FeII -NO) complex (120). The six-coordinate
FeII -NO complex is not stable and has only
been observed with time-resolved spectroscopic methods (120, 122, 123). The presence
of Mg2+ GTP or Mg2+ /cGMP/pyrophosphate
(PPi ) accelerates the formation of the
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five-coordinate sGC-NO complex, and
this effect is blocked by the addition of ATP
(122, 123).
Breakage of the Fe-His bond is thought to
be a critical step in the activation of sGC by
NO; however, recent data have shown that NO
coordination to the heme is not sufficient for
full activation ex vivo (122, 123). A low-activity
FeII -NO complex can be formed in the presence of stoichiometric amounts of NO, whereas
a high-activity FeII -NO complex is formed in
the presence of excess NO. These two fivecoordinate FeII -NO species are indistinguishable by electronic absorption spectroscopy, but
they exhibit distinct signals by electron paramagnetic spectroscopy (124). Preincubation of
sGC with substrate Mg2+ GTP or the reaction
products Mg2+ /cGMP/PPi produces a highactivity FeII -NO species in the presence of
stoichiometric amounts of NO (123). ATP can
compete with the GTP effect, and in the presence of ATP and GTP, a low-activity FeII -NO
species is formed (122). This clear difference
in sGC activity indicates that preincubation
of the enzyme with the small molecules leads
to a conformational change such that sGC is
highly activated by low levels of NO. It was
also found that the small-molecule YC-1 can
activate the low-activity FeII -NO complex to
the high-activity state (122, 125). Thus, both
YC-1 and excess NO activate the low-activity
five-coordinate FeII -NO complex that is
formed in the absence of substrate or reaction
products.
Two mechanisms have been proposed to account for the varying activity of the sGC FeII NO complex observed ex vivo. One proposal
is that excess NO activates the FeII -NO complex by binding to nonheme sites on the protein (122). If NO binds to a nonheme site,
it is likely that cysteines would comprise this
binding site as experiments with the thiol reactive reagent methyl methanethiosulfonate suggest that reduced cysteines are necessary for the
mechanism of NO activation (125). The second
proposal regarding sGC activation involves excess NO binding to the heme to form a transient dinitrosyl complex, which then converts
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In vitro measurements
NO dissociation
NO
NO
Slow
FeII
FeII
Preincubation with GTP increases rate
Slow
FeII
His
His
His
399 nm
399 nm
431 nm
ATP inhibits this GTP effect
YC-1 or BAY 41-2272 increases rate
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Deactivation
NO
NO
Fast
NO
FeII
FeII
His
His
High activity
No effect with GTP or ATP
YC-1 decreases rate
Low activity
Figure 5
Model of nitric oxide (NO) dissociation and deactivation of NO-stimulated soluble guanylate cyclase (sGC)
in vitro. The sGC-NO complex consists of two different five-coordinate species that slowly interconvert.
NO dissociation from the sGC heme is slow, whereas NO deactivation is rapid on the basis of in vitro
measurements, including spectroscopic methods (Absmax indicated in nanometers) and cyclic GMP analysis.
The presence of allosteric modulators (GTP, ATP, YC-1, or BAY 41-2272) influences the deactivation and
NO dissociation rates differently. Taken together, these results indicate that the deactivated sGC species is
distinct from the unligated sGC species (431 nm) produced from NO dissociation from the heme and
suggests that two molecules of NO are able to bind to sGC.
to a five-coordinate complex with NO bound in
the proximal heme pocket (123). Such proximal
heme pocket NO binding has been observed
with other histidine-ligated heme proteins including cytochrome c (126).
The importance of the two different sGCNO states in vivo remains unclear. One report
examining sGC activity in endothelial cells
proposed that enzyme activation by excess NO
is important for the physiological activation of
sGC (125), whereas another report examining
sGC activation in rat platelets and cerebellar
cells proposed that the observed activation
kinetics were consistent with a single NObinding event (127). Thus, further experiments
are necessary to resolve the mechanism of sGC
activation in vivo.
Deactivation and Nitric
Oxide Dissociation
Deactivation of the sGC FeII -NO complex
has been extensively studied to understand the
lifetime of the NO signal in vivo. This process
was originally thought to correlate directly with
NO dissociation from the sGC heme; however,
as mentioned above, a NO-bound protein that
is partially activated has been characterized.
This indicates that sGC deactivation and NO
dissociation from the heme cofactor are not
explicitly linked, and thus, deactivation and
heme-NO dissociation measurements cannot
be used interchangeably (Figure 5).
NO-induced relaxation of smooth muscle
cells dissipates within seconds upon removal of
free NO due to the deactivation of sGC (128).
sGC deactivation, the rate of decline in cGMP
synthesis after an activator is removed, has
been determined in various cells and with purified protein. In cells, sGC deactivation is rapid
(t1/2 < 5 s at 37◦ C) in the presence of an NO trap
(oxyhemoglobin) to limit NO rebinding (121,
129). Activity assays with the cytosol of retina
homogenate indicate that the sGC-NO deactivation rate is not influenced by the presence
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S-nitrosation:
posttranslational
modification involving
the oxidative addition
of NO to a thiol
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of Mg2+ GTP and is only slightly increased by
the presence of reducing agents, such as glutathione and dithiothreitol (129). In agreement
with cellular data, the deactivation rate of purified protein is also rapid (t1/2 ∼ 4 s at 37◦ C)
(130). This deactivation rate is unaffected by the
presence of a GTP analog and/or ATP (122),
but the rate is significantly decreased (140-fold)
by the presence of the allosteric activator YC-1
(t1/2 > 10 min at 37◦ C) (130).
The rate of NO dissociation from the sGC
heme must be spectroscopically determined,
and as a consequence, this rate has only been
measured in vitro. Early reports found that
the sGC heme-NO complex is very stable
(t1/2 ∼ 87 min at 37◦ C) in the absence of NO
traps or allosteric effectors (23, 131). Unlike
most heme proteins, which rapidly oxidize in
the presence of O2 and NO, a reduced FeII
protein is formed when NO dissociates from
the sGC heme. Using a trap to scavenge NO,
like CO/dithionite or oxymyoglobin, the dissociation rate can be determined without interference from NO rebinding. Reductants like
dithiothreitol or glutathione react with NO and
can also be used to prevent rebinding (23). With
these NO trapping methods, it was determined
that the NO dissociation rate is relatively slow
(t1/2 = 3–8 min at 37◦ C) (23, 27, 131) compared to previously determined deactivation
rates but increases (∼50-fold) in the presence
of Mg2+ GTP (131). However, if Mg2+ GTP is
added to sGC after the NO complex is formed,
it has no effect on the dissociation rate (23, 123,
131), indicating that the allosteric affect of GTP
is dependent on substrate binding in the absence of NO. In contrast to the previously discussed deactivation results, the GTP effect on
the dissociation rate is inhibited by the presence of ATP (122). Therefore, in the presence
of both GTP and ATP, sGC exhibits a slow
NO dissociation rate but rapid deactivation. In
the presence of YC-1, the NO dissociation rate
increases (27), whereas the deactivation rate decreases (130). A model summarizing deactivation of sGC and NO dissociation is shown in
Figure 5.
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Desensitization
The sGC response to GTN or NO decreases
upon repeated exposure to the activators. This
desensitization is rapid, such that a single
pretreatment of cells with GTN or NO can
significantly reduce cGMP stimulation upon
the second exposure (132, 133). Purified
protein in the absence of reducing agents also
exhibits a decrease in NO-stimulated activity
after pretreatment with NO. This desensitization has implications for therapies used in
the treatment of heart disease as this loss of
responsiveness, or tolerance, is also observed
when organic nitrates are administered to
patients with angina pectoris (134). As a consequence, these drugs cannot be used repeatedly.
Over the past 30 years, several proposals have
been advanced to explain the phenomenon of
tolerance, including increased PDE activity
(135), inhibition of mitochondrial aldehyde
dehydrogenase (136), and most recently
S-nitrosation (137, 138). There is currently
significant evidence in support of the proposal
that nitrosation, the oxidative addition of NO
to a thiol, contributes to sGC desensitization.
It has long been known that thiol oxidation
inhibits sGC activity. Cysteines can oxidize
sequentially to form a sulfenic acid, sulfinic
acid, sulfonic acid, or a disulfide bond in the
presence of O2 , or form a nitrosothiol in the
presence of NO and O2 . In the 1980s, the formation of sGC-cysteine mixed disulfides was
shown to reversibly inhibit cGMP production
(139). More recently, the induction of reactive
O2 species in vascular smooth muscle cells was
found to inhibit cGMP synthesis and lead to the
oxidative modification of sGC cysteines (140).
Additionally, the thiol modifying reagent
methyl methanethiosulfonate was shown to
inhibit sGC activity in vitro and in primary
endothelial cells. In vitro sGC inhibition by
methyl methanethiosulfonate is reversible, and
the molecule was shown to modify several
cysteines on both the α1 and β1 subunits
(125). The NO-dependent oxidation of cysteines on sGC has also been reported. sGC
is S-nitrosated in the presence of low levels
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of NO, and this modification has been linked
to a reduction in NO-stimulated activity
(137). In addition, GTN has been shown to
induce sGC S-nitrosation and desensitization
in a concentration-dependent manner (138).
Cysteines on both the α1 (C243) and β1
(C122) subunits have been identified as targets
of this oxidative modification.
It has become clear that sGC requires free
thiols for proper function. Without a crystal
structure, it is difficult to address why these cysteines are necessary for NO-induced sGC activation. Perhaps free thiols are important to the
structural integrity of the protein, involved in a
conformational change to the activated enzyme
state, and/or directly involved in the NO mechanism of activation. If sGC S-nitrosation is the
primary molecular mechanism of nitrate tolerance, then molecules developed to protect these
thiols from oxidation could be useful for the
treatment of diseases related to sGC dysfunction. Furthermore, the apo- and heme-oxidized
sGC states have been proposed to be physiologically relevant sGC species in diseased tissue.
MODULATORS OF SOLUBLE
GUANYLATE CYCLASE ACTIVITY
Soluble Guanylate Cyclase Activators
sGC is a therapeutic target in the treatment
of heart disease. Organic nitrites and organic
nitrates (like GTN) are perhaps the earliest
agents used to target the NO/cGMP-signaling
pathway and have been in clinical use for over
100 years. The first description of GTN as a
therapeutic agent for the treatment of angina
pectoris appeared in 1879, and it remains the
drug of choice to treat the disease. Despite
decades of clinical use, the precise mechanism
of action of GTN is unknown; however, it
is generally considered a nitrovasodilator or
a NO-donor. This NO release may occur by
spontaneous decomposition or bioconversion
to result in NO-dependent sGC activation.
In addition to NO, other known heme ligands, including CO, nitrosoalkanes, and alkyl
isocyanides, can bind to the sGC heme but can
only weakly activate the protein (17, 141, 142).
The binding of these compounds leads to the
formation of a six-coordinate complex and to a
two- to fourfold increase in the rate of cGMP
production, significantly lower than the 100- to
400-fold increase in cGMP synthesis observed
with NO. Other compounds that have been reported to activate sGC by targeting the hemebinding pocket include protoporphyrin IX (15)
and Co2+ protoporphyrin IX (143).
It has become clear that small molecules
can modulate the activity of sGC and that
new therapeutics might be developed for the
treatment of various diseases. This prompted
a search for novel sGC activators, and several
compounds were screened for the ability to increase cGMP levels in cell lysates. Such a screen
led to the identification of YC-1, a benzylindazole derivative that activates sGC without
coordinating to the heme (144). YC-1 only
activates the FeII -unligated sGC state two- to
fourfold but significantly increases sGC activity
when a ligand is bound at the FeII heme (141,
142, 145, 146). This synergistic activation leads
to an FeII -CO complex that is activated 100to 400-fold and an FeII -NO complex that is
activated 200- to 800-fold. The molecular
mechanism of YC-1 activation is unknown.
Experiments with equilibrium dialysis suggest
that sGC binds one equivalent of YC-1 per
heterodimer (147). This binding site may
be contained within the N terminus of the
α1 subunit (33, 148) or within the pseudosymmetric substrate site (149–151). What
is clear is that YC-1 binding to sGC induces
a conformational change that elevates cGMP
production. This highly active conformational
state has been characterized by spectroscopic
methods, including electronic absorption
(152), resonance Raman spectroscopy (147,
153), and electron paramagnetic resonance
spectroscopy (124, 147). There are also clear
kinetic effects of adding YC-1 to CO- or
NO-bound sGC (27, 130, 154).
The discovery of the novel sGC stimulator
YC-1 led several groups to carry out structure
activity relationships to improve both the solubility and efficacy of YC-1. With this work
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Nitrate tolerance:
loss of sensitivity to
nitrates
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came the identification of several other compounds, including BAY 41-2272, BAY 41-8543,
CMF-1571, and A-350619 (reviewed in Reference 103). Although there is some debate over
the possible inhibition of PDE5 by BAY 412272 (155, 156), it is commonly accepted that
the molecule activates sGC without coordinating to the heme. Collectively, these molecules
constitute a novel class of sGC stimulators
that require the presence of the heme moiety
and have the ability to synergistically stimulate
sGC with both NO and CO. These molecules
were promising drug candidates but had unfavorable drug metabolism. Medicinal chemistry efforts to reduce the problems associated
with these compounds led to the discovery of
BAY 63-2521 (riociguat). This compound induces vasodilation by activating sGC and is currently in Phase III clinical trails (reviewed in
Reference 157).
There are also small-molecule activators
that target heme-oxidized or -deficient sGC;
therefore, they are classified as NO and heme
independent. One of these is BAY 58-2667
(Cinaciguat) (158). BAY 58-2667 is generally
cited as an activator of both heme-deficient
and heme-oxidized (FeIII ) sGC (158), but there
is one report that proposes the molecule exclusively targets the heme-deficient sGC state
(159). BAY 58-2667 is proposed to activate sGC
by binding within the heme pocket, thereby
serving as a mimic of the heme-NO complex
(91, 101). In addition to activating the enzyme, BAY 58-2667 also stabilizes sGC and
protects the protein from degradation (160). In
rats, BAY 58-2667 was shown to lower blood
pressure (161) and protect against ischemic injury (162), and the compound has been tested
on patients with acute decompensated heart
failure (163–165).
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BI81CH22-Marletta
Soluble Guanylate Cyclase Inhibitors
There has been significantly more work on
identifying sGC activators, but compounds
that inhibit sGC activity have also been reported. These compounds are not as selective as the identified activators and include
general oxidants, molecules that target hemoproteins, and nucleotide cyclase inhibitors.
Compounds proposed to target the sGC HNOX domain include heme, hematin (15),
and 1H-[1,2,4]oxadiazolol[4,3-a]quinoxalin-1one (ODQ) (166). These compounds inhibit
sGC by oxidation of the ferrous iron in
the heme cofactor. Molecules such as hydrogen peroxide, superoxide, cystine, and Snitrosocysteine also reduce cGMP production
by oxidizing critical residues on sGC (138–
140). Other known sGC inhibitors, such as
LY83583 (167) and methylene blue (168), inhibit the enzyme via generation of superoxide anion (169, 170). Inhibition of sGC
by these oxidants emphasizes the importance
of the redox environment for proper protein
function.
Substrate analogs that bind to the sGC
catalytic domain inhibit guanylate cyclase
activity. To date, several such analogs have
been identified as sGC inhibitors, including,
but not limited to, ITP, XTP, CTP, ATP,
ADP, AMP, 2 -deoxyadenosine 5 -diphosphate
GDP,
guanosine-5 -[(α,β)(2 -dADP),
methylene]triphosphate (GMP-CPP), and
N-methylanthraniloyl (MANT)-nucleotides
(75, 151, 171, 172). These compounds inhibit
sGC by varying mechanisms depending on
whether they target the pseudosymmetric site
in addition to the substrate-binding site. For
example, AMP-PNP is a competitive inhibitor
(151), while ATP is a mixed-type inhibitor and
substrate (76).
SUMMARY POINTS
1. Soluble guanylate cyclase (sGC) is the most thoroughly characterized receptor for the
signaling molecule nitric oxide (NO). NO activation of sGC is essential to several physiological processes, and thus dysfunction in sGC is linked to several diseases. Compounds
that modulate sGC are in clinical trails for the treatment of heart disease.
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2. Each sGC subunit consists of an H-NOX, PAS, coiled-coil, and catalytic domain. Functional truncations of sGC include the isolated β1 H-NOX domain, the β1 coiled-coil
domain, and the α1β1 catalytic domains.
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3. Genome mining has revealed that several predicted proteins exist with high sequence
homology to sGC, including prokaryotic H-NOXs and eukaryotic NO-sensitive and
atypical sGCs.
4. The structure of full-length sGC is not yet determined, but studies on H-NOXs and sGC
homologs have illuminated many important structural features of the enzyme. On the
basis of these studies, sGC likely contains a hydrophobic heme-binding pocket, without
a distal-pocket hydrogen bond donor, and heterodimerization is mediated by contacts
on the PAS, coiled-coil, and catalytic domains.
5. α1β1 sGC does not form a stable complex with O2 , but several sGC homologs bind O2 .
Hydrogen bond donors in the heme-distal pocket are often found in the H-NOXs and
atypical sGCs that bind O2 . The presence or absence of amino acids capable of forming
a hydrogen bond in the heme-distal pocket contributes to selectivity for gaseous ligands.
6. sGC regulation by NO and nucleotides in vitro is complex. Two molecules of NO can
bind to the protein, and both GTP and ATP can influence NO binding, dissociation,
and activation. In cells, it is likely that NO, GTP, and ATP modulate cGMP production,
and both reduced cysteines and a reduced heme iron are important for this activity.
FUTURE ISSUES
Over the past few years, sGC has been implicated in an expanding number of physiological processes and diseases. Continued progress toward elucidating the mechanism of sGC
activation is important in the development of therapeutics to treat disorders relating to
the NO-signaling pathway. Despite significant advances in our understanding of NO as
a signaling agent, many questions remain unanswered. In mammals, the sGC response to
NO is complicated, and the precise events that lead to activation remain a topic of debate.
Furthermore, sGC is regulated by allosteric interactions with ATP and GTP, and there are
likely other important factors, including S-nitrosation and phosphorylation, that modulate
sGC activity. Future experiments considering these factors will be necessary to determine
the influence of each regulatory factor in the activation, deactivation, and desensitization of
sGC. How do the α1 and β1 subunits interact when sGC is activated? What is the nature
of the allosteric activator-binding site? What conformational changes occur when ligands
bind to sGC? All of these questions may be addressed with the structural elucidation of the
full-length heterodimeric protein. Additionally, structural analysis may provide insight into
the molecular mechanisms that lead to sGC dysfunction.
DISCLOSURE STATEMENT
M.A.M. is a cofounder of Omniox, Inc., a company commercializing oxygen delivery technology
for a broad range of clinical indications in cancer, surgical, and cardiovascular markets. The authors
are not aware of any other affiliations, memberships, funding, or financial holdings that might be
perceived as affecting the objectivity of this review.
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ACKNOWLEDGMENTS
This article was written while M.A.M. was employed by the University of California, Berkeley.
The authors acknowledge members of the Marletta lab, past and present, who have contributed to
studies on sGC, NOS, and H-NOXs. Research on sGC in the authors’ laboratory was supported
by the National Institutes of Health (NIH grant GM077365), and research on H-NOXs by NIH
grant GM070671.
LITERATURE CITED
Annu. Rev. Biochem. 2012.81:533-559. Downloaded from www.annualreviews.org
Access provided by b-on: Universidade Nova de Lisboa (UNL) on 01/18/15. For personal use only.
1. Brunton TL. 1867. On the use of nitrite of amyl in angina pectoris. Lancet 90:97–98
2. Murrell W. 1879. Nitro-glycerine as a remedy for angina pectoris. Lancet 113:113–15
3. Garbers DL, Chrisman TD, Wiegn P, Katafuchi T, Albanesi JP, et al. 2006. Membrane guanylyl cyclase
receptors: an update. Trends Endocrinol. Metab. 17:251–58
4. Padayatti PS, Pattanaik P, Ma X, van den Akker F. 2004. Structural insights into the regulation and the
activation mechanism of mammalian guanylyl cyclases. Pharmacol. Ther. 104:83–99
5. Feussner M, Richter H, Baum O, Gossrau R. 2001. Association of soluble guanylate cyclase with the
sarcolemma of mammalian skeletal muscle fibers. Acta Histochem. 103:265–77
6. Russwurm M, Wittau N, Koesling D. 2001. Guanylyl cyclase/PSD-95 interaction: targeting of the nitric
oxide-sensitive α2β1 guanylyl cyclase to synaptic membranes. J. Biol. Chem. 276:44647–52
7. Zabel U, Kleinschnitz C, Oh P, Nedvetsky P, Smolenski A, et al. 2002. Calcium-dependent membrane
association sensitizes soluble guanylyl cyclase to nitric oxide. Nat. Cell Biol. 4:307–11
8. Madhani M, Okorie M, Hobbs AJ, MacAllister RJ. 2006. Reciprocal regulation of human soluble and
particulate guanylate cyclases in vivo. Br. J. Pharmacol. 149:797–801
9. Munzel T, Feil R, Mulsch A, Lohmann SM, Hofmann F, Walter U. 2003. Physiology and pathophysiology of vascular signaling controlled by guanosine 3 ,5 -cyclic monophosphate-dependent protein kinase.
Circulation 108:2172–83
10. Sanders KM, Ward SM, Thornbury KD, Dalziel HH, Westfall DP, Carl A. 1992. Nitric oxide as
a non-adrenergic, non-cholinergic neurotransmitter in the gastrointestinal tract. Jap. J. Pharmacol.
58(Suppl.):P220–25
11. Warner TD, Mitchell JA, Sheng H, Murad F. 1994. Effects of cyclic GMP on smooth muscle relaxation.
Adv. Pharmacol. 26:171–94
12. Garbers DL. 1979. Purification of soluble guanylate cyclase from rat lung. J. Biol. Chem. 254:240–43
13. Braughler JM, Mittal CK, Murad F. 1979. Purification of soluble guanylate cyclase from rat liver. Proc.
Natl. Acad. Sci. USA 76:219–22
14. Gerzer R, Hofmann F, Schultz G. 1981. Purification of a soluble, sodium-nitroprusside-stimulated
guanylate cyclase from bovine lung. Eur. J. Biochem. 116:479–86
15. Ignarro LJ, Wood KS, Wolin MS. 1982. Activation of purified soluble guanylate cyclase by protoporphyrin IX. Proc. Natl. Acad. Sci. USA 79:2870–73
16. Vogel KM, Hu S, Spiro TG, Dierks EA, Yu AE, Burstyn JN. 1999. Variable forms of soluble guanylyl
cyclase: protein-ligand interactions and the issue of activation by carbon monoxide. J. Biol. Inorg. Chem.
4:804–13
17. Stone JR, Marletta MA. 1994. Soluble guanylate cyclase from bovine lung: activation with nitric oxide and
carbon monoxide and spectral characterization of the ferrous and ferric states. Biochemistry 33:5636–40
18. Russwurm M, Koesling D. 2005. Purification and characterization of NO-sensitive guanylyl cyclase.
Methods Enzymol. 396:492–501
19. Harteneck C, Koesling D, Soling A, Schultz G, Bohme E. 1990. Expression of soluble guanylyl cyclase.
Catalytic activity requires two enzyme subunits. FEBS Lett. 272:221–23
20. Buechler WA, Nakane M, Murad F. 1991. Expression of soluble guanylate cyclase activity requires both
enzyme subunits. Biochem. Biophys. Res. Commun. 174:351–57
21. Friebe A, Wedel B, Harteneck C, Foerster J, Schultz G, Koesling D. 1997. Functions of conserved
cysteines of soluble guanylyl cyclase. Biochemistry 36:1194–98
552
Derbyshire
·
Marletta
Annu. Rev. Biochem. 2012.81:533-559. Downloaded from www.annualreviews.org
Access provided by b-on: Universidade Nova de Lisboa (UNL) on 01/18/15. For personal use only.
BI81CH22-Marletta
ARI
3 May 2012
12:17
22. Zhou Z, Gross S, Roussos C, Meurer S, Muller-Esterl W, Papapetropoulos A. 2004. Structural and functional characterization of the dimerization region of soluble guanylyl cyclase. J. Biol. Chem. 279:24935–43
23. Brandish PE, Buechler W, Marletta MA. 1998. Regeneration of the ferrous heme of soluble guanylate
cyclase from the nitric oxide complex: acceleration by thiols and oxyhemoglobin. Biochemistry 37:16898–
907
24. Hoenicka M, Becker EM, Apeler H, Sirichoke T, Schroder H, et al. 1999. Purified soluble guanylyl
cyclase expressed in a baculovirus/Sf9 system: stimulation by YC-1, nitric oxide, and carbon monoxide.
J. Mol. Med. 77:14–23
25. Lee YC, Martin E, Murad F. 2000. Human recombinant soluble guanylyl cyclase: expression, purification,
and regulation. Proc. Natl. Acad. Sci. USA 97:10763–68
26. Wagner C, Russwurm M, Jager R, Friebe A, Koesling D. 2005. Dimerization of nitric oxide-sensitive
guanylyl cyclase requires the α1 N terminus. J. Biol. Chem. 280:17687–93
27. Winger JA, Derbyshire ER, Marletta MA. 2007. Dissociation of nitric oxide from soluble guanylate
cyclase and heme-nitric oxide/oxygen binding domain constructs. J. Biol. Chem. 282:897–907
28. Karow DS, Pan D, Davis JH, Behrends S, Mathies RA, Marletta MA. 2005. Characterization of functional
heme domains from soluble guanylate cyclase. Biochemistry 44:16266–74
29. Zhao Y, Marletta MA. 1997. Localization of the heme binding region in soluble guanylate cyclase.
Biochemistry 36:15959–64
30. Zhong F, Pan J, Liu X, Wang H, Ying T, et al. 2011. A novel insight into the heme and NO/CO binding
mechanism of the alpha subunit of human soluble guanylate cyclase. J. Biol. Inorg. Chem. 16:1227–39
31. Ma X, Beuve A, van den Akker F. 2010. Crystal structure of the signaling helix coiled-coil domain of the
β1 subunit of the soluble guanylyl cyclase. BMC Struct. Biol. 10:2
32. Winger JA, Marletta MA. 2005. Expression and characterization of the catalytic domains of soluble
guanylate cyclase: interaction with the heme domain. Biochemistry 44:4083–90
33. Hu X, Murata LB, Weichsel A, Brailey JL, Roberts SA, et al. 2008. Allostery in recombinant soluble
guanylyl cyclase from Manduca sexta. J. Biol. Chem. 283:20968–77
34. Harteneck C, Wedel B, Koesling D, Malkewitz J, Bohme E, Schultz G. 1991. Molecular cloning and
expression of a new α-subunit of soluble guanylyl cyclase. Interchangeability of the α-subunits of the
enzyme. FEBS Lett. 292:217–22
35. Yuen PS, Potter LR, Garbers DL. 1990. A new form of guanylyl cyclase is preferentially expressed in
rat kidney. Biochemistry 29:10872–78
36. Budworth J, Meillerais S, Charles I, Powell K. 1999. Tissue distribution of the human soluble guanylate
cyclases. Biochem. Biophys. Res. Commun. 263:696–701
37. Bellingham M, Evans TJ. 2007. The α2β1 isoform of guanylyl cyclase mediates plasma membrane
localized nitric oxide signalling. Cell Signal. 19:2183–93
38. Russwurm M, Behrends S, Harteneck C, Koesling D. 1998. Functional properties of a naturally occurring
isoform of soluble guanylyl cyclase. Biochem J. 335(Part 1):125–30
39. Behrends S, Harteneck C, Schultz G, Koesling D. 1995. A variant of the α2 subunit of soluble guanylyl
cyclase contains an insert homologous to a region within adenylyl cyclases and functions as a dominant
negative protein. J. Biol. Chem. 270:21109–13
40. Neitz A, Mergia E, Eysel UT, Koesling D, Mittmann T. 2011. Presynaptic nitric oxide/cGMP facilitates
glutamate release via hyperpolarization-activated cyclic nucleotide-gated channels in the hippocampus.
Eur. J. Neurosci. 33:1611–21
41. Koglin M, Vehse K, Budaeus L, Scholz H, Behrends S. 2001. Nitric oxide activates the β2 subunit of
soluble guanylyl cyclase in the absence of a second subunit. J. Biol. Chem. 276:30737–43
42. Behrends S, Budaeus L, Kempfert J, Scholz H, Starbatty J, Vehse K. 2001. The β2 subunit of nitric
oxide-sensitive guanylyl cyclase is developmentally regulated in rat kidney. Naunyn-Schmiedebergs Arch.
Pharmacol. 364:573–76
43. Friebe A, Koesling D. 2009. The function of NO-sensitive guanylyl cyclase: What we can learn from
genetic mouse models. Nitric Oxide 21:149–56
44. Friebe A, Mergia E, Dangel O, Lange A, Koesling D. 2007. Fatal gastrointestinal obstruction and
hypertension in mice lacking nitric oxide-sensitive guanylyl cyclase. Proc. Natl. Acad. Sci. USA 104:7699–
704
www.annualreviews.org • Structure and Regulation of SGC
553
ARI
3 May 2012
12:17
45. Groneberg D, Konig P, Wirth A, Offermanns S, Koesling D, Friebe A. 2010. Smooth muscle-specific
deletion of nitric oxide-sensitive guanylyl cyclase is sufficient to induce hypertension in mice. Circulation
121:401–9
46. Haghikia A, Mergia E, Friebe A, Eysel UT, Koesling D, Mittmann T. 2007. Long-term potentiation in
the visual cortex requires both nitric oxide receptor guanylyl cyclases. J. Neurosci. 27:818–23
47. Mergia E, Friebe A, Dangel O, Russwurm M, Koesling D. 2006. Spare guanylyl cyclase NO receptors
ensure high NO sensitivity in the vascular system. J. Clin. Investig. 116:1731–37
48. Nimmegeers S, Sips P, Buys E, Brouckaert P, Van de Voorde J. 2007. Functional role of the soluble
guanylyl cyclase α1 subunit in vascular smooth muscle relaxation. Cardiovasc. Res. 76:149–59
49. Vermeersch P, Buys E, Pokreisz P, Marsboom G, Ichinose F, et al. 2007. Soluble guanylate cyclaseα1 deficiency selectively inhibits the pulmonary vasodilator response to nitric oxide and increases the
pulmonary vascular remodeling response to chronic hypoxia. Circulation 116:936–43
50. Vanneste G, Dhaese I, Sips P, Buys E, Brouckaert P, Lefebvre RA. 2007. Gastric motility in soluble
guanylate cyclase α1 knock-out mice. J. Physiol. 584:907–20
51. Dhaese I, Vanneste G, Sips P, Buys E, Brouckaert P, Lefebvre RA. 2008. Involvement of soluble guanylate
cyclase α1 and α2 , and SKCa channels in NANC relaxation of mouse distal colon. Eur. J. Pharmacol.
589:251–59
52. Tinette S, Zhang L, Garnier A, Engler G, Tares S, Robichon A. 2007. Exploratory behaviour in NOdependent cyclase mutants of Drosophila shows defects in coincident neuronal signalling. BMC Neurosci.
8:65
53. Gibbs SM, Becker A, Hardy RW, Truman JW. 2001. Soluble guanylate cyclase is required during
development for visual system function in Drosophila. J. Neurosci. 21:7705–14
54. Riedl CA, Neal SJ, Robichon A, Westwood JT, Sokolowski MB. 2005. Drosophila soluble guanylyl
cyclase mutants exhibit increased foraging locomotion: behavioral and genomic investigations. Behav.
Genet. 35:231–44
55. Zayas RM, Trimmer BA. 2007. Characterization of NO/cGMP-mediated responses in identified motoneurons. Cell Mol. Neurobiol. 27:191–209
56. Wilson CH, Christensen TA, Nighorn AJ. 2007. Inhibition of nitric oxide and soluble guanylyl cyclase
signaling affects olfactory neuron activity in the moth, Manduca sexta. J. Comp. Physiol. A 193:715–28
57. Fujie S, Yamamoto T, Murakami J, Hatakeyama D, Shiga H, et al. 2005. Nitric oxide synthase and
soluble guanylyl cyclase underlying the modulation of electrical oscillations in a central olfactory organ.
J. Neurobiol. 62:14–30
58. Gelperin A, Flores J, Raccuia-Behling F, Cooke IR. 2000. Nitric oxide and carbon monoxide modulate
oscillations of olfactory interneurons in a terrestrial mollusk. J. Neurophysiol. 83:116–27
59. Cary SP, Winger JA, Derbyshire ER, Marletta MA. 2006. Nitric oxide signaling: no longer simply on
or off. Trends Biochem. Sci. 31:231–39
60. Wedel B, Humbert P, Harteneck C, Foerster J, Malkewitz J, et al. 1994. Mutation of His-105 in the
β1 subunit yields a nitric oxide-insensitive form of soluble guanylyl cyclase. Proc. Natl. Acad. Sci. USA
91:2592–96
61. Wedel B, Harteneck C, Foerster J, Friebe A, Schultz G, Koesling D. 1995. Functional domains of soluble
guanylyl cyclase. J. Biol. Chem. 270:24871–75
62. Iyer LM, Anantharaman V, Aravind L. 2003. Ancient conserved domains shared by animal soluble
guanylyl cyclases and bacterial signaling proteins. BMC Genomics 4:5
63. Nioche P, Berka V, Vipond J, Minton N, Tsai AL, Raman CS. 2004. Femtomolar sensitivity of a NO
sensor from Clostridium botulinum. Science 306:1550–53
64. Pellicena P, Karow DS, Boon EM, Marletta MA, Kuriyan J. 2004. Crystal structure of an oxygen-binding
heme domain related to soluble guanylate cyclases. Proc. Natl. Acad. Sci. USA 101:12854–59
65. Boon EM, Marletta MA. 2005. Ligand discrimination in soluble guanylate cyclase and the H-NOX
family of heme sensor proteins. Curr. Opin. Chem. Biol. 9:441–46
66. Moglich A, Ayers RA, Moffat K. 2009. Structure and signaling mechanism of Per-ARNT-Sim domains.
Structure 17:1282–94
67. Shiga T, Suzuki N. 2005. Amphipathic α-helix mediates the heterodimerization of soluble guanylyl
cyclase. Zool. Sci. 22:735–42
Annu. Rev. Biochem. 2012.81:533-559. Downloaded from www.annualreviews.org
Access provided by b-on: Universidade Nova de Lisboa (UNL) on 01/18/15. For personal use only.
BI81CH22-Marletta
554
Derbyshire
·
Marletta
BI81CH22-Marletta
ARI
3 May 2012
12:17
68. Rothkegel C, Schmidt PM, Atkins DJ, Hoffmann LS, Schmidt HH, et al. 2007. Dimerization region of soluble guanylate cyclase characterized by bimolecular fluorescence complementation in vivo.
Mol. Pharmacol. 72:1181–90
69. Ma X, Sayed N, Baskaran P, Beuve A, van den Akker F. 2008. PAS-mediated dimerization of soluble
guanylyl cyclase revealed by signal transduction histidine kinase domain crystal structure. J. Biol. Chem.
283:1167–78
70. Tesmer JJ, Sunahara RK, Gilman AG, Sprang SR. 1997. Crystal structure of the catalytic domains of
adenylyl cyclase in a complex with Gsα GTPγS. Science 278:1907–16
71. Zhang G, Liu Y, Ruoho AE, Hurley JH. 1997. Structure of the adenylyl cyclase catalytic core. Nature
386:247–53
72. Sunahara RK, Beuve A, Tesmer JJ, Sprang SR, Garbers DL, Gilman AG. 1998. Exchange of substrate
and inhibitor specificities between adenylyl and guanylyl cyclases. J. Biol. Chem. 273:16332–38
73. Brandwein HJ, Lewicki JA, Waldman SA, Murad F. 1982. Effect of GTP analogues on purified soluble
guanylate cyclase. J. Biol. Chem. 257:1309–11
74. Garbers DL, Suddath JL, Hardman JG. 1975. Enzymatic formation of inosine 3 ,5 -monophosphate
and of 2 -deoxyguanosine 3 ,5 -monophosphate. Inosinate and deoxyguanylate cyclase activity. Biochim.
Biophys. Acta 377:174–85
75. Gille A, Lushington GH, Mou TC, Doughty MB, Johnson RA, Seifert R. 2004. Differential inhibition
of adenylyl cyclase isoforms and soluble guanylyl cyclase by purine and pyrimidine nucleotides. J. Biol.
Chem. 279:19955–69
76. Derbyshire ER, Fernhoff NB, Deng S, Marletta MA. 2009. Nucleotide regulation of soluble guanylate
cyclase substrate specificity. Biochemistry 48:7519–24
77. Haase T, Haase N, Kraehling JR, Behrends S. 2010. Fluorescent fusion proteins of soluble guanylyl
cyclase indicate proximity of the heme nitric oxide domain and catalytic domain. PLoS ONE 5:e11617
78. Winger JA, Derbyshire ER, Lamers MH, Marletta MA, Kuriyan J. 2008. The crystal structure of the
catalytic domain of a eukaryotic guanylate cyclase. BMC Struct. Biol. 8:42
79. Rauch A, Leipelt M, Russwurm M, Steegborn C. 2008. Crystal structure of the guanylyl cyclase Cya2.
Proc. Natl. Acad. Sci. USA 105:15720–25
80. Morton DB. 2004. Atypical soluble guanylyl cyclases in Drosophila can function as molecular oxygen
sensors. J. Biol. Chem. 279:50651–53
81. Morton DB, Langlais KK, Stewart JA, Vermehren A. 2005. Comparison of the properties of the five
soluble guanylyl cyclase subunits in Drosophila melanogaster. J. Insect Sci. 5:12
82. Huang SH, Rio DC, Marletta MA. 2007. Ligand binding and inhibition of an oxygen-sensitive soluble
guanylate cyclase, Gyc-88E, from Drosophila. Biochemistry 46:15115–22
83. Vermehren-Schmaedick A, Ainsley JA, Johnson WA, Davies SA, Morton DB. 2010. Behavioral responses
to hypoxia in Drosophila larvae are mediated by atypical soluble guanylyl cyclases. Genetics 186:183–96
84. Gray JM, Karow DS, Lu H, Chang AJ, Chang JS, et al. 2004. Oxygen sensation and social feeding
mediated by a C. elegans guanylate cyclase homologue. Nature 430:317–22
85. Cheung BH, Arellano-Carbajal F, Rybicki I, de Bono M. 2004. Soluble guanylate cyclases act in neurons
exposed to the body fluid to promote C. elegans aggregation behavior. Curr. Biol. 14:1105–11
86. Cheung BH, Cohen M, Rogers C, Albayram O, de Bono M. 2005. Experience-dependent modulation
of C. elegans behavior by ambient oxygen. Curr. Biol. 15:905–17
87. Zimmer M, Gray JM, Pokala N, Chang AJ, Karow DS, et al. 2009. Neurons detect increases and decreases
in oxygen levels using distinct guanylate cyclases. Neuron 61:865–79
88. Price MS, Chao LY, Marletta MA. 2007. Shewanella oneidensis MR-1 H-NOX regulation of a histidine
kinase by nitric oxide. Biochemistry 46:13677–83
89. Wang Y, Dufour YS, Carlson HK, Donohue TJ, Marletta MA, Ruby EG. 2010. H-NOX-mediated nitric
oxide sensing modulates symbiotic colonization by Vibrio fischeri. Proc. Natl. Acad. Sci. USA 107:8375–80
90. Boon EM, Huang SH, Marletta MA. 2005. A molecular basis for NO selectivity in soluble guanylate
cyclase. Nat. Chem. Biol. 1:53–59
91. Schmidt PM, Schramm M, Schroder H, Wunder F, Stasch JP. 2004. Identification of residues crucially
involved in the binding of the heme moiety of soluble guanylate cyclase. J. Biol. Chem. 279:3025–32
Annu. Rev. Biochem. 2012.81:533-559. Downloaded from www.annualreviews.org
Access provided by b-on: Universidade Nova de Lisboa (UNL) on 01/18/15. For personal use only.
·
www.annualreviews.org • Structure and Regulation of SGC
555
ARI
3 May 2012
12:17
92. Zhao Y, Schelvis JP, Babcock GT, Marletta MA. 1998. Identification of histidine 105 in the β1 subunit
of soluble guanylate cyclase as the heme proximal ligand. Biochemistry 37:4502–9
93. Ma X, Sayed N, Beuve A, van den Akker F. 2007. NO and CO differentially activate soluble guanylyl
cyclase via a heme pivot-bend mechanism. EMBO J. 26:578–88
94. Erbil WK, Price MS, Wemmer DE, Marletta MA. 2009. A structural basis for H-NOX signaling in
Shewanella oneidensis by trapping a histidine kinase inhibitory conformation. Proc. Natl. Acad. Sci. USA
106:19753–60
95. Tran R, Boon EM, Marletta MA, Mathies RA. 2009. Resonance Raman spectra of an O2 -binding HNOX domain reveal heme relaxation upon mutation. Biochemistry 48:8568–77
96. Ibrahim M, Derbyshire ER, Marletta MA, Spiro TG. 2010. Probing soluble guanylate cyclase activation
by CO and YC-1 using resonance Raman spectroscopy. Biochemistry 49:3815–23
97. Ibrahim M, Derbyshire ER, Soldatova AV, Marletta MA, Spiro TG. 2010. Soluble guanylate cyclase is
activated differently by excess NO and by YC-1: resonance Raman spectroscopic evidence. Biochemistry
49:4864–71
98. Pal B, Tanaka K, Takenaka S, Kitagawa T. 2010. Resonance Raman spectroscopic investigation of
structural changes of CO-heme in soluble guanylate cyclase generated by effectors and substrate.
J. Raman Spectrosc. 41:1178–84
99. Olea C, Boon EM, Pellicena P, Kuriyan J, Marletta MA. 2008. Probing the function of heme distortion
in the H-NOX family. ACS Chem. Biol. 3:703–10
100. Tsai AL, Berka V, Martin F, Ma X, van den Akker F, et al. 2010. Is Nostoc H-NOX a NO sensor or redox
switch? Biochemistry 49:6587–99
101. Martin F, Baskaran P, Ma X, Dunten PW, Schaefer M, et al. 2010. Structure of cinaciguat (BAY 58-2667)
bound to Nostoc H-NOX domain reveals insights into heme-mimetic activation of the soluble guanylyl
cyclase. J. Biol. Chem. 285:22651–57
102. Barrick D. 1994. Replacement of the proximal ligand of sperm whale myoglobin with free imidazole in
the mutant His-93→Gly. Biochemistry 33:6546–54
103. Evgenov OV, Pacher P, Schmidt PM, Hasko G, Schmidt HH, Stasch JP. 2006. NO-independent stimulators and activators of soluble guanylate cyclase: discovery and therapeutic potential. Nat. Rev. Drug
Discov. 5:755–68
104. Baskaran P, Heckler EJ, van den Akker F, Beuve A. 2011. Aspartate 102 in the heme domain of soluble
guanylyl cyclase has a key role in NO activation. Biochemistry 50:4291–97
105. Derbyshire ER, Deng S, Marletta MA. 2010. Incorporation of tyrosine and glutamine residues into the
soluble guanylate cyclase heme distal pocket alters NO and O2 binding. J. Biol. Chem. 285:17471–78
106. Rothkegel C, Schmidt PM, Stoll F, Schroder H, Schmidt HHHW, Stasch JP. 2006. Identification of
residues crucially involved in soluble guanylate cyclase activation. FEBS Lett. 580:4205–13
107. Capece L, Estrin DA, Marti MA. 2008. Dynamical characterization of the heme NO oxygen binding
(HNOX) domain. Insight into soluble guanylate cyclase allosteric transition. Biochemistry 47:9416–27
108. Xu C, Ibrahim M, Spiro TG. 2008. DFT analysis of axial and equatorial effects on heme-CO vibrational
modes: applications to CooA and H-NOX heme sensor proteins. Biochemistry 47:2379–87
109. Makino R, Park SY, Obayashi E, Iizuka T, Hori H, Shiro Y. 2011. Oxygen binding and redox properties
of the heme in soluble guanylate cyclase: implications for the mechanism of ligand discrimination.
J. Biol. Chem. 286:15678–87
110. Deinum G, Stone JR, Babcock GT, Marletta MA. 1996. Binding of nitric oxide and carbon monoxide
to soluble guanylate cyclase as observed with resonance Raman spectroscopy. Biochemistry 35:1540–7
111. Jain R, Chan MK. 2003. Mechanisms of ligand discrimination by heme proteins. J. Biol. Inorg. Chem.
8:1–11
112. Zhang Y, Lu M, Cheng Y, Li Z. 2010. H-NOX domains display different tunnel systems for ligand
migration. J. Mol. Graph. Model. 28:814–19
113. Winter MB, Herzik MA, Kuriyan J, Marletta MA. 2011. Tunnels modulate ligand flux in a heme nitric
oxide/oxygen binding (H-NOX) domain. Proc. Natl. Acad. Sci. USA. 108:E881–89
114. Martin E, Berka V, Bogatenkova E, Murad F, Tsai AL. 2006. Ligand selectivity of soluble guanylyl
cyclase: effect of the hydrogen-bonding tyrosine in the distal heme pocket on binding of oxygen, nitric
oxide, and carbon monoxide. J. Biol. Chem. 281:27836–45
Annu. Rev. Biochem. 2012.81:533-559. Downloaded from www.annualreviews.org
Access provided by b-on: Universidade Nova de Lisboa (UNL) on 01/18/15. For personal use only.
BI81CH22-Marletta
556
Derbyshire
·
Marletta
Annu. Rev. Biochem. 2012.81:533-559. Downloaded from www.annualreviews.org
Access provided by b-on: Universidade Nova de Lisboa (UNL) on 01/18/15. For personal use only.
BI81CH22-Marletta
ARI
3 May 2012
12:17
115. Das TK, Samuni U, Lin Y, Goldberg DE, Rousseau DL, Friedman JM. 2004. Distal heme pocket
conformers of carbonmonoxy derivatives of Ascaris hemoglobin: evidence of conformational trapping in
porous sol-gel matrices. J. Biol. Chem. 279:10433–41
116. Giangiacomo L, Ilari A, Boffi A, Morea V, Chiancone E. 2005. The truncated oxygen-avid hemoglobin
from Bacillus subtilis: X-ray structure and ligand binding properties. J. Biol. Chem. 280:9192–202
117. Marti MA, Capece L, Bikiel DE, Falcone B, Estrin DA. 2007. Oxygen affinity controlled by dynamical distal conformations: the soybean leghemoglobin and the Paramecium caudatum hemoglobin cases.
Proteins 68:480–87
118. Milani M, Pesce A, Ouellet Y, Dewilde S, Friedman J, et al. 2004. Heme-ligand tunneling in group I
truncated hemoglobins. J. Biol. Chem. 279:21520–25
119. Schrammel A, Behrends S, Schmidt K, Koesling D, Mayer B. 1996. Characterization of 1H[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one as a heme-site inhibitor of nitric oxide-sensitive guanylyl cyclase. Mol. Pharmacol. 50:1–5
120. Stone JR, Marletta MA. 1996. Spectral and kinetic studies on the activation of soluble guanylate cyclase
by nitric oxide. Biochemistry 35:1093–99
121. Bellamy TC, Garthwaite J. 2001. Sub-second kinetics of the nitric oxide receptor, soluble guanylyl
cyclase, in intact cerebellar cells. J. Biol. Chem. 276:4287–92
122. Cary SP, Winger JA, Marletta MA. 2005. Tonic and acute nitric oxide signaling through soluble guanylate
cyclase is mediated by nonheme nitric oxide, ATP, and GTP. Proc. Natl. Acad. Sci. USA 102:13064–69
123. Russwurm M, Koesling D. 2004. NO activation of guanylyl cyclase. EMBO J. 23:4443–50
124. Derbyshire ER, Gunn A, Ibrahim M, Spiro TG, Britt RD, Marletta MA. 2008. Characterization of two
different five-coordinate soluble guanylate cyclase ferrous-nitrosyl complexes. Biochemistry 47:3892–99
125. Fernhoff NB, Derbyshire ER, Marletta MA. 2009. A nitric oxide/cysteine interaction mediates the
activation of soluble guanylate cyclase. Proc. Natl. Acad. Sci. USA 106:21602–7
126. Lawson DM, Stevenson CE, Andrew CR, Eady RR. 2000. Unprecedented proximal binding of nitric
oxide to heme: implications for guanylate cyclase. EMBO J. 19:5661–71
127. Roy B, Garthwaite J. 2006. Nitric oxide activation of guanylyl cyclase in cells revisited. Proc. Natl. Acad.
Sci. USA 103:12185–90
128. Palmer RM, Ferrige AG, Moncada S. 1987. Nitric oxide release accounts for the biological activity of
endothelium-derived relaxing factor. Nature 327:524–26
129. Margulis A, Sitaramayya A. 2000. Rate of deactivation of nitric oxide-stimulated soluble guanylate cyclase:
influence of nitric oxide scavengers and calcium. Biochemistry 39:1034–39
130. Russwurm M, Mergia E, Mullershausen F, Koesling D. 2002. Inhibition of deactivation of NO-sensitive
guanylyl cyclase accounts for the sensitizing effect of YC-1. J. Biol. Chem. 277:24883–88
131. Kharitonov VG, Russwurm M, Magde D, Sharma VS, Koesling D. 1997. Dissociation of nitric oxide
from soluble guanylate cyclase. Biochem. Biophys. Res. Commun. 239:284–86
132. Bellamy TC, Wood J, Goodwin DA, Garthwaite J. 2000. Rapid desensitization of the nitric oxide
receptor, soluble guanylyl cyclase, underlies diversity of cellular cGMP responses. Proc. Natl. Acad. Sci.
USA 97:2928–33
133. Schroder H, Leitman DC, Bennett BM, Waldman SA, Murad F. 1988. Glyceryl trinitrate-induced
desensitization of guanylate cyclase in cultured rat lung fibroblasts. J. Pharmacol. Exp. Ther. 245:413–18
134. Elkayam U, Kulick D, McIntosh N, Roth A, Hsueh W, Rahimtoola SH. 1987. Incidence of early tolerance
to hemodynamic effects of continuous infusion of nitroglycerin in patients with coronary artery disease
and heart failure. Circulation 76:577–84
135. Kim D, Rybalkin SD, Pi X, Wang Y, Zhang C, et al. 2001. Upregulation of phosphodiesterase 1A1
expression is associated with the development of nitrate tolerance. Circulation 104:2338–43
136. Sydow K, Daiber A, Oelze M, Chen Z, August M, et al. 2004. Central role of mitochondrial aldehyde
dehydrogenase and reactive oxygen species in nitroglycerin tolerance and cross-tolerance. J. Clin. Investig.
113:482–89
137. Sayed N, Baskaran P, Ma X, van den Akker F, Beuve A. 2007. Desensitization of soluble guanylyl cyclase,
the NO receptor, by S-nitrosylation. Proc. Natl. Acad. Sci. USA 104:12312–17
www.annualreviews.org • Structure and Regulation of SGC
557
ARI
3 May 2012
12:17
138. Sayed N, Kim DD, Fioramonti X, Iwahashi T, Duran WN, Beuve A. 2008. Nitroglycerin-induced Snitrosylation and desensitization of soluble guanylyl cyclase contribute to nitrate tolerance. Circ. Res.
103:606–14
139. Brandwein HJ, Lewicki JA, Murad F. 1981. Reversible inactivation of guanylate cyclase by mixed disulfide
formation. J. Biol. Chem. 256:2958–62
140. Maron BA, Zhang YY, Handy DE, Beuve A, Tang SS, et al. 2009. Aldosterone increases oxidant stress
to impair guanylyl cyclase activity by cysteinyl thiol oxidation in vascular smooth muscle cells. J. Biol.
Chem. 284:7665–72
141. Derbyshire ER, Marletta MA. 2007. Butyl isocyanide as a probe of the activation mechanism of soluble
guanylate cyclase. Investigating the role of non-heme nitric oxide. J. Biol. Chem. 282:35741–48
142. Derbyshire ER, Tran R, Mathies RA, Marletta MA. 2005. Characterization of nitrosoalkane binding
and activation of soluble guanylate cyclase. Biochemistry 44:16257–65
143. Makino R, Matsuda H, Obayashi E, Shiro Y, Iizuka T, Hori H. 1999. EPR characterization of axial bond
in metal center of native and cobalt-substituted guanylate cyclase. J. Biol. Chem. 274:7714–23
144. Ko FN, Wu CC, Kuo SC, Lee FY, Teng CM. 1994. YC-1, a novel activator of platelet guanylate cyclase.
Blood 84:4226–33
145. Friebe A, Schultz G, Koesling D. 1996. Sensitizing soluble guanylyl cyclase to become a highly COsensitive enzyme. EMBO J. 15:6863–68
146. Stone JR, Marletta MA. 1998. Synergistic activation of soluble guanylate cyclase by YC-1 and carbon
monoxide: implications for the role of cleavage of the iron-histidine bond during activation by nitric
oxide. Chem. Biol. 5:255–61
147. Makino R, Obayashi E, Homma N, Shiro Y, Hori H. 2003. YC-1 facilitates release of the proximal His
residue in the NO and CO complexes of soluble guanylate cyclase. J. Biol. Chem. 278:11130–37
148. Stasch JP, Becker EM, Alonso-Alija C, Apeler H, Dembowsky K, et al. 2001. NO-independent regulatory
site on soluble guanylate cyclase. Nature 410:212–15
149. Friebe A, Russwurm M, Mergia E, Koesling D. 1999. A point-mutated guanylyl cyclase with features of
the YC-1-stimulated enzyme: implications for the YC-1 binding site? Biochemistry 38:15253–57
150. Lamothe M, Chang FJ, Balashova N, Shirokov R, Beuve A. 2004. Functional characterization of nitric
oxide and YC-1 activation of soluble guanylyl cyclase: structural implication for the YC-1 binding site?
Biochemistry 43:3039–48
151. Yazawa S, Tsuchiya H, Hori H, Makino R. 2006. Functional characterization of two nucleotide-binding
sites in soluble guanylate cyclase. J. Biol. Chem. 281:21763–70
152. Kharitonov VG, Sharma VS, Magde D, Koesling D. 1999. Kinetics and equilibria of soluble guanylate
cyclase ligation by CO: effect of YC-1. Biochemistry 38:10699–706
153. Denninger JW, Schelvis JP, Brandish PE, Zhao Y, Babcock GT, Marletta MA. 2000. Interaction of
soluble guanylate cyclase with YC-1: kinetic and resonance Raman studies. Biochemistry 39:4191–98
154. Sharma VS, Magde D, Kharitonov VG, Koesling D. 1999. Soluble guanylate cyclase: effect of YC-1 on
ligation kinetics with carbon monoxide. Biochem. Biophys. Res. Commun. 254:188–91
155. Bischoff E, Stasch JP. 2004. Effects of the sGC stimulator BAY 41-2272 are not mediated by phosphodiesterase 5 inhibition. Circulation 110:e320–21; author reply e20–1
156. Mullershausen F, Russwurm M, Friebe A, Koesling D. 2004. Inhibition of phosphodiesterase type 5 by
the activator of nitric oxide-sensitive guanylyl cyclase BAY 41-2272. Circulation 109:1711–13
157. Belik J. 2009. Riociguat, an oral soluble guanylate cyclase stimulator for the treatment of pulmonary
hypertension. Curr. Opin. Investig. Drugs 10:971–79
158. Stasch JP, Schmidt P, Alonso-Alija C, Apeler H, Dembowsky K, et al. 2002. NO- and haem-independent
activation of soluble guanylyl cyclase: molecular basis and cardiovascular implications of a new pharmacological principle. Br. J. Pharmacol. 136:773–83
159. Roy B, Mo E, Vernon J, Garthwaite J. 2008. Probing the presence of the ligand-binding haem in cellular
nitric oxide receptors. Br. J. Pharmacol. 153:1495–504
160. Meurer S, Pioch S, Pabst T, Opitz N, Schmidt PM, et al. 2009. Nitric oxide-independent vasodilator
rescues heme-oxidized soluble guanylate cyclase from proteasomal degradation. Circ. Res. 105:33–41
161. Kalk P, Godes M, Relle K, Rothkegel C, Hucke A, et al. 2006. NO-independent activation of soluble
guanylate cyclase prevents disease progression in rats with 5/6 nephrectomy. Br. J. Pharmacol. 148:853–59
Annu. Rev. Biochem. 2012.81:533-559. Downloaded from www.annualreviews.org
Access provided by b-on: Universidade Nova de Lisboa (UNL) on 01/18/15. For personal use only.
BI81CH22-Marletta
558
Derbyshire
·
Marletta
Annu. Rev. Biochem. 2012.81:533-559. Downloaded from www.annualreviews.org
Access provided by b-on: Universidade Nova de Lisboa (UNL) on 01/18/15. For personal use only.
BI81CH22-Marletta
ARI
3 May 2012
12:17
162. Korkmaz S, Radovits T, Barnucz E, Hirschberg K, Neugebauer P, et al. 2009. Pharmacological activation
of soluble guanylate cyclase protects the heart against ischemic injury. Circulation 120:677–86
163. Frey R, Muck W, Unger S, Artmeier-Brandt U, Weimann G, Wensing G. 2008. Pharmacokinetics,
pharmacodynamics, tolerability, and safety of the soluble guanylate cyclase activator cinaciguat (BAY
58-2667) in healthy male volunteers. J. Clin. Pharmacol. 48:1400–10
164. Lapp H, Mitrovic V, Franz N, Heuer H, Buerke M, et al. 2009. Cinaciguat (BAY 58-2667) improves cardiopulmonary hemodynamics in patients with acute decompensated heart failure. Circulation 119:2781–
88
165. Mueck W, Frey R. 2010. Population pharmacokinetics and pharmacodynamics of cinaciguat, a soluble
guanylate cyclase activator, in patients with acute decompensated heart failure. Clin. Pharmacokinet.
49:119–29
166. Garthwaite J, Southam E, Boulton CL, Nielsen EB, Schmidt K, Mayer B. 1995. Potent and selective inhibition of nitric oxide-sensitive guanylyl cyclase by 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one.
Mol. Pharmacol. 48:184–88
167. Mulsch A, Busse R, Liebau S, Forstermann U. 1988. LY 83583 interferes with the release of endotheliumderived relaxing factor and inhibits soluble guanylate cyclase. J. Pharmacol. Exp. Ther. 247:283–88
168. Gruetter CA, Kadowitz PJ, Ignarro LJ. 1981. Methylene blue inhibits coronary arterial relaxation and
guanylate cyclase activation by nitroglycerin, sodium nitrite, and amyl nitrite. Can. J. Physiol. Pharmacol.
59:150–56
169. Lee YS, Wurster RD. 1995. Mechanism of potentiation of LY83583-induced growth inhibition by
sodium nitroprusside in human brain tumor cells. Cancer Chemother. Pharmacol. 36:341–44
170. Wolin MS, Cherry PD, Rodenburg JM, Messina EJ, Kaley G. 1990. Methylene blue inhibits vasodilation
of skeletal muscle arterioles to acetylcholine and nitric oxide via the extracellular generation of superoxide
anion. J. Pharmacol. Exp. Ther. 254:872–76
171. Chang FJ, Lemme S, Sun Q, Sunahara RK, Beuve A. 2005. Nitric oxide-dependent allosteric inhibitory
role of a second nucleotide binding site in soluble guanylyl cyclase. J. Biol. Chem. 280:11513–19
172. Suzuki T, Suematsu M, Makino R. 2001. Organic phosphates as a new class of soluble guanylate cyclase
inhibitors. FEBS Lett. 507:49–53
www.annualreviews.org • Structure and Regulation of SGC
559
BI81-FrontMatter
ARI
9 May 2012
7:26
Annu. Rev. Biochem. 2012.81:533-559. Downloaded from www.annualreviews.org
Access provided by b-on: Universidade Nova de Lisboa (UNL) on 01/18/15. For personal use only.
Contents
Annual Review of
Biochemistry
Volume 81, 2012
Preface
Preface and Dedication to Christian R.H. Raetz
JoAnne Stubbe p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p pxi
Prefatories
A Mitochondrial Odyssey
Walter Neupert p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 1
The Fires of Life
Gottfried Schatz p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p34
Chromatin, Epigenetics, and Transcription Theme
Introduction to Theme “Chromatin, Epigenetics, and Transcription”
Joan W. Conaway p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p61
The COMPASS Family of Histone H3K4 Methylases:
Mechanisms of Regulation in Development
and Disease Pathogenesis
Ali Shilatifard p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p65
Programming of DNA Methylation Patterns
Howard Cedar and Yehudit Bergman p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p97
RNA Polymerase II Elongation Control
Qiang Zhou, Tiandao Li, and David H. Price p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 119
Genome Regulation by Long Noncoding RNAs
John L. Rinn and Howard Y. Chang p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 145
Protein Tagging Theme
The Ubiquitin System, an Immense Realm
Alexander Varshavsky p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 167
Ubiquitin and Proteasomes in Transcription
Fuqiang Geng, Sabine Wenzel, and William P. Tansey p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 177
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The Ubiquitin Code
David Komander and Michael Rape p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 203
Ubiquitin and Membrane Protein Turnover: From Cradle to Grave
Jason A. MacGurn, Pi-Chiang Hsu, and Scott D. Emr p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 231
The N-End Rule Pathway
Takafumi Tasaki, Shashikanth M. Sriram, Kyong Soo Park, and Yong Tae Kwon p p p 261
Annu. Rev. Biochem. 2012.81:533-559. Downloaded from www.annualreviews.org
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Ubiquitin-Binding Proteins: Decoders of Ubiquitin-Mediated
Cellular Functions
Koraljka Husnjak and Ivan Dikic p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 291
Ubiquitin-Like Proteins
Annemarthe G. van der Veen and Hidde L. Ploegh p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 323
Recent Advances in Biochemistry
Toward the Single-Hour High-Quality Genome
Patrik L. Ståhl and Joakim Lundeberg p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 359
Mass Spectrometry–Based Proteomics and Network Biology
Ariel Bensimon, Albert J.R. Heck, and Ruedi Aebersold p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 379
Membrane Fission: The Biogenesis of Transport Carriers
Felix Campelo and Vivek Malhotra p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 407
Emerging Paradigms for Complex Iron-Sulfur Cofactor Assembly
and Insertion
John W. Peters and Joan B. Broderick p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 429
Structural Perspective of Peptidoglycan Biosynthesis and Assembly
Andrew L. Lovering, Susan S. Safadi, and Natalie C.J. Strynadka p p p p p p p p p p p p p p p p p p p p 451
Discovery, Biosynthesis, and Engineering of Lantipeptides
Patrick J. Knerr and Wilfred A. van der Donk p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 479
Regulation of Glucose Transporter Translocation
in Health and Diabetes
Jonathan S. Bogan p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 507
Structure and Regulation of Soluble Guanylate Cyclase
Emily R. Derbyshire and Michael A. Marletta p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 533
The MPS1 Family of Protein Kinases
Xuedong Liu and Mark Winey p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 561
The Structural Basis for Control of Eukaryotic Protein Kinases
Jane A. Endicott, Martin E.M. Noble, and Louise N. Johnson p p p p p p p p p p p p p p p p p p p p p p p p p p 587
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Measurements and Implications of the Membrane Dipole Potential
Liguo Wang p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 615
GTPase Networks in Membrane Traffic
Emi Mizuno-Yamasaki, Felix Rivera-Molina, and Peter Novick p p p p p p p p p p p p p p p p p p p p p p p 637
Roles for Actin Assembly in Endocytosis
Olivia L. Mooren, Brian J. Galletta, and John A. Cooper p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 661
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Lipid Droplets and Cellular Lipid Metabolism
Tobias C. Walther and Robert V. Farese Jr. p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 687
Adipogenesis: From Stem Cell to Adipocyte
Qi Qun Tang and M. Daniel Lane p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 715
Pluripotency and Nuclear Reprogramming
Marion Dejosez and Thomas P. Zwaka p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 737
Endoplasmic Reticulum Stress and Type 2 Diabetes
Sung Hoon Back and Randal J. Kaufman p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 767
Structure Unifies the Viral Universe
Nicola G.A. Abrescia, Dennis H. Bamford, Jonathan M. Grimes,
and David I. Stuart p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 795
Indexes
Cumulative Index of Contributing Authors, Volumes 77–81 p p p p p p p p p p p p p p p p p p p p p p p p p p p 823
Cumulative Index of Chapter Titles, Volumes 77–81 p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 827
Errata
An online log of corrections to Annual Review of Biochemistry articles may be found at
http://biochem.annualreviews.org/errata.shtml
Contents
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Annual Reviews
It’s about time. Your time. It’s time well spent.
New From Annual Reviews:
Annual Review of Statistics and Its Application
Volume 1 • Online January 2014 • http://statistics.annualreviews.org
Annu. Rev. Biochem. 2012.81:533-559. Downloaded from www.annualreviews.org
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Editor: Stephen E. Fienberg, Carnegie Mellon University
Associate Editors: Nancy Reid, University of Toronto
Stephen M. Stigler, University of Chicago
The Annual Review of Statistics and Its Application aims to inform statisticians and quantitative methodologists, as
well as all scientists and users of statistics about major methodological advances and the computational tools that
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Complimentary online access to the first volume will be available until January 2015.
table of contents:
• What Is Statistics? Stephen E. Fienberg
• A Systematic Statistical Approach to Evaluating Evidence
from Observational Studies, David Madigan, Paul E. Stang,
Jesse A. Berlin, Martijn Schuemie, J. Marc Overhage,
Marc A. Suchard, Bill Dumouchel, Abraham G. Hartzema,
Patrick B. Ryan
• High-Dimensional Statistics with a View Toward Applications
in Biology, Peter Bühlmann, Markus Kalisch, Lukas Meier
• Next-Generation Statistical Genetics: Modeling, Penalization,
and Optimization in High-Dimensional Data, Kenneth Lange,
Jeanette C. Papp, Janet S. Sinsheimer, Eric M. Sobel
• The Role of Statistics in the Discovery of a Higgs Boson,
David A. van Dyk
• Breaking Bad: Two Decades of Life-Course Data Analysis
in Criminology, Developmental Psychology, and Beyond,
Elena A. Erosheva, Ross L. Matsueda, Donatello Telesca
• Brain Imaging Analysis, F. DuBois Bowman
• Event History Analysis, Niels Keiding
• Statistics and Climate, Peter Guttorp
• Statistical Evaluation of Forensic DNA Profile Evidence,
Christopher D. Steele, David J. Balding
• Climate Simulators and Climate Projections,
Jonathan Rougier, Michael Goldstein
• Probabilistic Forecasting, Tilmann Gneiting,
Matthias Katzfuss
• Bayesian Computational Tools, Christian P. Robert
• Bayesian Computation Via Markov Chain Monte Carlo,
Radu V. Craiu, Jeffrey S. Rosenthal
• Build, Compute, Critique, Repeat: Data Analysis with Latent
Variable Models, David M. Blei
• Structured Regularizers for High-Dimensional Problems:
Statistical and Computational Issues, Martin J. Wainwright
• Using League Table Rankings in Public Policy Formation:
Statistical Issues, Harvey Goldstein
• Statistical Ecology, Ruth King
• Estimating the Number of Species in Microbial Diversity
Studies, John Bunge, Amy Willis, Fiona Walsh
• Dynamic Treatment Regimes, Bibhas Chakraborty,
Susan A. Murphy
• Statistics and Related Topics in Single-Molecule Biophysics,
Hong Qian, S.C. Kou
• Statistics and Quantitative Risk Management for Banking
and Insurance, Paul Embrechts, Marius Hofert
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