Structure and Activation of Soluble Guanylyl Cyclase, the Nitric Oxide Sensor

Montfort, W.R.; Wales, Jessica A.; Weichsel, A.

doi:10.1089/ars.2016.6693

Cited by 116 publications

(105 citation statements)

References 125 publications

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“…For activation, binding of stimulatory compounds to the N‐terminal β1 H‐NOX domain must lead to optimal alignment of the two C‐terminal catalytic domains, which together form the active site . Possible mechanisms for this range from large‐scale conformational changes that release inhibitory domain contacts to subtle through‐protein adjustments that permit optimal cyclase domain alignment . Here, using a truncated form of sGC lacking cyclase domains, we show that the central coiled‐coil helix regulates heme affinity for CO and appears to undergo a change in conformation upon CO binding that can be blocked by disulfide bond formation.…”

Section: Discussionmentioning

confidence: 99%

Instability in a coiled‐coil signaling helix is conserved for signal transduction in soluble guanylyl cyclase

et al. 2019

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How nitric oxide (NO) activates its primary receptor, α1/β1 soluble guanylyl cyclase (sGC or GC‐1), remains unknown. Likewise, how stimulatory compounds enhance sGC activity is poorly understood, hampering development of new treatments for cardiovascular disease. NO binding to ferrous heme near the N‐terminus in sGC activates cyclase activity near the C‐terminus, yielding cGMP production and physiological response. CO binding can also stimulate sGC, but only weakly in the absence of stimulatory small‐molecule compounds, which together lead to full activation. How ligand binding enhances catalysis, however, has yet to be discovered. Here, using a truncated version of sGC from Manduca sexta, we demonstrate that the central coiled‐coil domain, the most highly conserved region of the ~150,000 Da protein, not only provides stability to the heterodimer but is also conformationally active in signal transduction. Sequence conservation in the coiled coil includes the expected heptad‐repeating pattern for coiled‐coil motifs, but also invariant positions that disfavor coiled‐coil stability. Full‐length coiled coil dampens CO affinity for heme, while shortening of the coiled coil leads to enhanced CO binding. Introducing double mutation αE447L/βE377L, predicted to replace two destabilizing glutamates with leucines, lowers CO binding affinity while increasing overall protein stability. Likewise, introduction of a disulfide bond into the coiled coil results in reduced CO affinity. Taken together, we demonstrate that the heme domain is greatly influenced by coiled‐coil conformation, suggesting communication between heme and catalytic domains is through the coiled coil. Highly conserved structural imperfections in the coiled coil provide needed flexibility for signal transduction.

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Section: Discussionmentioning

confidence: 99%

Instability in a coiled‐coil signaling helix is conserved for signal transduction in soluble guanylyl cyclase

et al. 2019

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“…7120 (32,33) and Shewanella oneidensis (34). These structures display the same overall fold and provide a solid scaffold for understanding H-NOX structure in sGC function (reviewed in (4,5,23)). The overall H-NOX fold is ~180 residues long and displays an N-terminal sub-domain encompassing residues 1-60, which is dominated by a 3-helix bundle, followed by a larger mixed helix/sheet sub-domain that contains the heme pocket.…”

Section: Characterization Of Compound Binding By Transferred Noesy Nmmentioning

confidence: 99%

“…Central to NO signaling is soluble guanylyl cyclase (sGC), the NO receptor, which regulates vascular tone, platelet activation, wound healing and other factors of importance to cardiovascular health (4,5). sGC is an ~150 kDa heterodimeric NO sensor composed of a and b subunits, with an a1/b1 isoform predominating in vascular tissue (also referred to as guanylyl cyclase-1, GC-1), and an a2/b1 isoform predominating in nerve cells (called GC-2), where it is important for memory formation.…”

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confidence: 99%

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Discovery of stimulator binding to a conserved pocket in the heme domain of soluble guanylyl cyclase

Wales

Chen

Breci

et al. 2018

Journal of Biological Chemistry

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Soluble guanylyl cyclase (sGC) is the receptor for nitric oxide and a highly soughtafter therapeutic target for the management of cardiovascular diseases. New compounds that stimulate sGC show clinical promise, but where these stimulator compounds bind and how they function remains unknown. Here, using a photolyzable diazirine derivative of a novel stimulator compound, IWP-051, and MS analysis, we localized drug binding to the β1 heme domain of sGC proteins from the hawkmoth Manduca sexta and from human. Covalent attachments to the stimulator were also identified in bacterial homologs of the sGC heme domain, referred to as H-NOX domains, including those from Nostoc sp. 7120, Shewanella oneidensis, Shewanella woodyi, and Clostridium botulinum, indicating the binding site is highly conserved. The identification of photoaffinity-labeled peptides was aided by a signature MS fragmentation pattern of general applicability for unequivocal identification of covalently attached compounds. Using NMR, we also examined stimulator binding to sGC from M. sexta and bacterial H-NOX homologs. These data indicated that stimulators bind to a conserved cleft between two subdomains in the sGC heme domain. L23W/T48W substitutions within the binding pocket resulted in a 9-fold decrease in drug response, suggesting the bulkier tryptophan residues directly block stimulator binding.

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“…Soluble guanylyl cyclase (GC-1) maintains vascular function through the NO/GC-1/cGMP pathway [1,2] by catalyzing the conversion of GTP into cGMP (Fig. 1).…”

Section: Introductionmentioning

confidence: 99%

Structure/function of the soluble guanylyl cyclase catalytic domain

Childers

Garcin

2018

Nitric Oxide

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Soluble guanylyl cyclase (GC-1) is the primary receptor of nitric oxide (NO) in smooth muscle cells and maintains vascular function by inducing vasorelaxation in nearby blood vessels. GC-1 converts guanosine 5′-triphosphate (GTP) into cyclic guanosine 3′,5′-monophosphate (cGMP), which acts as a second messenger to improve blood flow. While much work has been done to characterize this pathway, we lack a mechanistic understanding of how NO binding to the heme domain leads to a large increase in activity at the C-terminal catalytic domain. Recent structural evidence and activity measurements from multiple groups have revealed a low-activity cyclase domain that requires additional GC-1 domains to promote a catalytically-competent conformation. How the catalytic domain structurally transitions into the active conformation requires further characterization. This review focuses on structure/function studies of the GC-1 catalytic domain and recent advances various groups have made in understanding how catalytic activity is regulated including small molecules interactions, Cys-S-NO modifications and potential interactions with the NO-sensor domain and other proteins.

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Structure and Activation of Soluble Guanylyl Cyclase, the Nitric Oxide Sensor

Abstract: Recent results and ongoing studies lay the foundation for a complete understanding of structure and mechanism, and they open the door for new drug discovery targeting sGC. Antioxid. Redox Signal. 26, 107-121.

Cited by 116 publications

References 125 publications

Instability in a coiled‐coil signaling helix is conserved for signal transduction in soluble guanylyl cyclase

Instability in a coiled‐coil signaling helix is conserved for signal transduction in soluble guanylyl cyclase

Discovery of stimulator binding to a conserved pocket in the heme domain of soluble guanylyl cyclase

Structure/function of the soluble guanylyl cyclase catalytic domain

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