Crystal structure of the Alpha subunit PAS domain from soluble guanylyl cyclase

Purohit, R.K.; Weichsel, A.; Montfort, W.R.

doi:10.1002/pro.2331

Cited by 44 publications

(40 citation statements)

References 38 publications

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“…In sGC, multiple protein domains must transfer signals to each other via direct and allosteric signaling mechanism involving many regulatory interfaces. In addition to the heme-based NO-binding site on the H-NOX domain and the active site in the catalytic domain, it has been suggested that sGC has an additional NO-binding site, an allosteric nucleotide-binding site in the catalytic domain (31), a site for heme-independent small-molecule activator binding [such as YC-1 (5-[1-(phenylmethyl)-1H-indazol-3-yl]-2-furanmethanol; 32, 33)], as well as an internal cavity in the PAS domain that may bind ligands (7). All of these sites must be accessible to allow modulation of the sGC catalytic activity through integration of the external signals.…”

Section: Discussionmentioning

confidence: 99%

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Single-particle EM reveals the higher-order domain architecture of soluble guanylate cyclase

Campbell

Underbakke

Potter

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

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Soluble guanylate cyclase (sGC) is the primary nitric oxide (NO) receptor in mammals and a central component of the NO-signaling pathway. The NO-signaling pathways mediate diverse physiological processes, including vasodilation, neurotransmission, and myocardial functions. sGC is a heterodimer assembled from two homologous subunits, each comprised of four domains. Although crystal structures of isolated domains have been reported, no structure is available for full-length sGC. We used single-particle electron microscopy to obtain the structure of the complete sGC heterodimer and determine its higher-order domain architecture. Overall, the protein is formed of two rigid modules: the catalytic dimer and the clustered Per/Art/Sim and heme-NO/O 2 -binding domains, connected by a parallel coiled coil at two hinge points. The quaternary assembly demonstrates a very high degree of flexibility. We captured hundreds of individual conformational snapshots of free sGC, NO-bound sGC, and guanosine-5′-[(α,β)-methylene]triphosphate-bound sGC. The molecular architecture and pronounced flexibility observed provides a significant step forward in understanding the mechanism of NO signaling.N itric oxide (NO) has emerged as an integral signaling molecule in biology. Soluble guanylate cyclase (sGC), the primary receptor of NO in mammals, binds NO via an Fe II heme cofactor leading to a several hundred-fold increase in 3,5-cyclic guanosine monophosphate (cGMP) synthesis. cGMP then acts as a second messenger, targeting phosphodiesterases, ion-gated channels, and cGMP-dependent protein kinases. These target proteins go on to regulate many critical physiological functions including vasodilation, platelet aggregation, neurotransmission, and myocardial functions (1, 2). Disruptions in NO signaling have been linked to hypertension, erectile dysfunction, neurodegeneration, stroke, and heart disease (3, 4). sGC has been the focus of small-molecule modulators of activity for therapeutic advantage. Riociguat, which is a stimulator of sGC, has recently been approved for treatment of pulmonary hypertension (5). However, the mechanistic details underlying the modulation of sGC catalytic activity by NO and other small molecules remain largely unknown. Determining the structure of the full-length sGC, free and in complex with NO, is therefore a prerequisite to understanding its function and for the design and improvement of therapeutics for treatment of diseases involving the NO/cGMP pathway.The most extensively studied and physiologically relevant isoform of sGC is the 150-kDa heterodimer containing one α1 and one β1 subunit. Each subunit is comprised of four modular domains: the N-terminal heme-NO/O 2 -binding (H-NOX), the Per/Arnt/Sim (PAS), the helical, and the C-terminal catalytic domain (Fig. 1). No structure of the complete holoenzyme is available to date, and its absence precludes answering key questions such as how NO occupancy of the N-terminal β H-NOX sensor domain is communicated to the C-terminal cyclase domain. Atomic models of isola...

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Section: Discussionmentioning

confidence: 99%

“…No structure of the complete holoenzyme is available to date, and its absence precludes answering key questions such as how NO occupancy of the N-terminal β H-NOX sensor domain is communicated to the C-terminal cyclase domain. Atomic models of isolated sGC domains have been obtained by X-ray crystallography or homology modeling (6)(7)(8)(9) (Fig. 1).…”

mentioning

confidence: 99%

Single-particle EM reveals the higher-order domain architecture of soluble guanylate cyclase

Campbell

Underbakke

Potter

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

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“…Consequently, it was concluded that NOsGC stimulators bind to the HNOX domain of ␤ 1 . Recent evidence indicates that ciguat stimulators relieve allosteric inhibition of the HNOX domain of ␤ 1 by the ␣ 1 PAS domain thereby enhancing NO binding (26,30). The differential influence of the ␣ 1 S-W466F mutation on ciguat/NO stimulation can be accommodated in such an allostery model where signal transmission from the HNOX domain of ␤ 1 via the ␣ 1 PAS domain and the coiled-coil region of ␣ 1 to the catalytic domain is not unidirectional.…”

Section: Discussionmentioning

confidence: 99%

Nitric Oxide Activation of Guanylate Cyclase Pushes the α1 Signaling Helix and the β1 Heme-binding Domain Closer to the Substrate-binding Site

Busker

Neidhardt

Behrends

2014

Journal of Biological Chemistry

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Background: NO-induced conformational changes of guanylate cyclase were analyzed. Results: FRET experiments show a movement of two tryptophans toward a fluorescent substrate. Conclusion: Activation is transmitted to the catalytic domain by direct interaction of the heme binding domain and through the coiled-coil helix. Significance: The results help to advance understanding of the molecular events associated with activation of NOsGC.

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“…The sGC␤1 PAS domain was modeled as a monomer and dimer using PAS domains from Manduca sexta sGC␣ and signal transduction histidine kinase of Nostoc punctiforme sp. PCC73102 as templates (37,38). We modeled a short segment of the coiled-coil domain of human sGC␤1 based on the nearly identical equivalent in rat sGC␤1 (39).…”

Section: Hydrogen/deuterium Exchange Reactions Of Individual Proteinsmentioning

confidence: 99%

“…Fig. 6C illustrates our exchange results in the context of a structural model for sGC␤1(1-359)-H105F that is based on existing structural and biophysical studies (37,38,43,57).…”

Section: Impact Of Complex Formation On Sgc␤1(1-359)-h105f-mentioning

confidence: 99%

Heat Shock Protein 90 Associates with the Per-Arnt-Sim Domain of Heme-free Soluble Guanylate Cyclase

Sarkar

Dai

Haque

et al. 2015

Journal of Biological Chemistry

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Background: hsp90␤ binds to heme-free sGC␤1 subunit to enable heme insertion during maturation, but the mechanism is unclear. Results: Their interaction involves the hsp90␤ M domain and the Per-Arnt-Sim (PAS) domain of apo-sGC␤1. Conclusion: hsp90␤ may modulate the apo-sGC␤1 through its PAS domain, suggesting a means to aid heme insertion. Significance: This work expands our knowledge of how hsp90 regulates PAS-containing client proteins like sGC.

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Crystal structure of the Alpha subunit PAS domain from soluble guanylyl cyclase

Cited by 44 publications

References 38 publications

Single-particle EM reveals the higher-order domain architecture of soluble guanylate cyclase

Single-particle EM reveals the higher-order domain architecture of soluble guanylate cyclase

Nitric Oxide Activation of Guanylate Cyclase Pushes the α1 Signaling Helix and the β1 Heme-binding Domain Closer to the Substrate-binding Site

Heat Shock Protein 90 Associates with the Per-Arnt-Sim Domain of Heme-free Soluble Guanylate Cyclase

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