Replacement of heme by soluble guanylate cyclase (sGC) activators abolishes heme-nitric oxide/oxygen (H-NOX) domain structural plasticity

The gasotransmitter nitric oxide (NO) is a critical endogenous regulator of homeostasis, in major part via the generation of cGMP (cyclic guanosine monophosphate) from GTP (guanosine triphosphate) by NO's main physiological receptor, the soluble guanylate cyclase (sGC). sGC is a heterodimer, co...

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Autores principales:	Aikaterini I. Argyriou, Garyfallia I. Makrynitsa, Georgios Dalkas, Dimitra A. Georgopoulou, Konstantinos Salagiannis, Vassiliki Vazoura, Andreas Papapetropoulos, Stavros Topouzis, Georgios A. Spyroulias
Formato:	article
Lenguaje:	EN
Publicado:	Elsevier 2021
Materias:	Soluble guanylate cyclase (sGC) Heme BAY 58-2667 BAY 60-2770 Cinaciguat H-NOX Biology (General) QH301-705.5
Acceso en línea:	https://doaj.org/article/cace431cf9e043178301ce8c6e55771f
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spelling	oai:doaj.org-article:cace431cf9e043178301ce8c6e55771f2021-12-02T05:03:40ZReplacement of heme by soluble guanylate cyclase (sGC) activators abolishes heme-nitric oxide/oxygen (H-NOX) domain structural plasticity2665-928X10.1016/j.crstbi.2021.11.003https://doaj.org/article/cace431cf9e043178301ce8c6e55771f2021-01-01T00:00:00Zhttp://www.sciencedirect.com/science/article/pii/S2665928X2100026Xhttps://doaj.org/toc/2665-928XThe gasotransmitter nitric oxide (NO) is a critical endogenous regulator of homeostasis, in major part via the generation of cGMP (cyclic guanosine monophosphate) from GTP (guanosine triphosphate) by NO's main physiological receptor, the soluble guanylate cyclase (sGC). sGC is a heterodimer, composed of an α1 and a β1 subunit, of which the latter contains the heme-nitric oxide/oxygen (H-NOX) domain, responsible for NO recognition, binding and signal initiation. The NO/sGC/cGMP axis is dysfunctional in a variety of diseases, including hypertension and heart failure, especially since oxidative stress results in heme oxidation, sGC unresponsiveness to NO and subsequent degradation. As a central player in this axis, sGC is the focus of intense research efforts aiming to develop therapeutic molecules that enhance its activity. A class of drugs named sGC “activators” aim to replace the oxidized heme of the H-NOX domain, thus stabilizing the enzyme and restoring its activity. Although numerous studies outline the pharmacology and binding behavior of these compounds, the static 3D models available so far do not allow a satisfactory understanding of the structural basis of sGC's activation mechanism by these drugs. Herein, application NMR describes different conformational states during the replacement of the heme by a sGC activators. We show that the two sGC activators (BAY 58-2667 and BAY 60-2770) significantly decrease the conformational plasticity of the recombinant H-NOX protein domain of Nostoc sp. cyanobacterium, rendering it a lot more rigid compared to the heme-occupied H-NOX. NMR methodology also reveals, for the first time, a surprising bi-directional competition between reduced heme and these compounds, pointing to a highly dynamic regulation of the H-NOX domain. This competitive, bi-directional mode of interaction is also confirmed by monitoring cGMP generation in A7r5 vascular smooth muscle cells by these activators. We show that, surprisingly, heme's redox state impacts differently the bioactivity of these two structurally similar compounds. In all, by NMR-based and functional approaches we contribute unique experimental insight into the dynamic interaction of sGC activators with the H-NOX domain and its dependence on the heme redox status, with the ultimate goal to permit a better design of such therapeutically important molecules.Aikaterini I. ArgyriouGaryfallia I. MakrynitsaGeorgios DalkasDimitra A. GeorgopoulouKonstantinos SalagiannisVassiliki VazouraAndreas PapapetropoulosStavros TopouzisGeorgios A. SpyrouliasElsevierarticleSoluble guanylate cyclase (sGC)HemeBAY 58-2667BAY 60-2770CinaciguatH-NOXBiology (General)QH301-705.5ENCurrent Research in Structural Biology, Vol 3, Iss , Pp 324-336 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Soluble guanylate cyclase (sGC) Heme BAY 58-2667 BAY 60-2770 Cinaciguat H-NOX Biology (General) QH301-705.5
spellingShingle	Soluble guanylate cyclase (sGC) Heme BAY 58-2667 BAY 60-2770 Cinaciguat H-NOX Biology (General) QH301-705.5 Aikaterini I. Argyriou Garyfallia I. Makrynitsa Georgios Dalkas Dimitra A. Georgopoulou Konstantinos Salagiannis Vassiliki Vazoura Andreas Papapetropoulos Stavros Topouzis Georgios A. Spyroulias Replacement of heme by soluble guanylate cyclase (sGC) activators abolishes heme-nitric oxide/oxygen (H-NOX) domain structural plasticity
description	The gasotransmitter nitric oxide (NO) is a critical endogenous regulator of homeostasis, in major part via the generation of cGMP (cyclic guanosine monophosphate) from GTP (guanosine triphosphate) by NO's main physiological receptor, the soluble guanylate cyclase (sGC). sGC is a heterodimer, composed of an α1 and a β1 subunit, of which the latter contains the heme-nitric oxide/oxygen (H-NOX) domain, responsible for NO recognition, binding and signal initiation. The NO/sGC/cGMP axis is dysfunctional in a variety of diseases, including hypertension and heart failure, especially since oxidative stress results in heme oxidation, sGC unresponsiveness to NO and subsequent degradation. As a central player in this axis, sGC is the focus of intense research efforts aiming to develop therapeutic molecules that enhance its activity. A class of drugs named sGC “activators” aim to replace the oxidized heme of the H-NOX domain, thus stabilizing the enzyme and restoring its activity. Although numerous studies outline the pharmacology and binding behavior of these compounds, the static 3D models available so far do not allow a satisfactory understanding of the structural basis of sGC's activation mechanism by these drugs. Herein, application NMR describes different conformational states during the replacement of the heme by a sGC activators. We show that the two sGC activators (BAY 58-2667 and BAY 60-2770) significantly decrease the conformational plasticity of the recombinant H-NOX protein domain of Nostoc sp. cyanobacterium, rendering it a lot more rigid compared to the heme-occupied H-NOX. NMR methodology also reveals, for the first time, a surprising bi-directional competition between reduced heme and these compounds, pointing to a highly dynamic regulation of the H-NOX domain. This competitive, bi-directional mode of interaction is also confirmed by monitoring cGMP generation in A7r5 vascular smooth muscle cells by these activators. We show that, surprisingly, heme's redox state impacts differently the bioactivity of these two structurally similar compounds. In all, by NMR-based and functional approaches we contribute unique experimental insight into the dynamic interaction of sGC activators with the H-NOX domain and its dependence on the heme redox status, with the ultimate goal to permit a better design of such therapeutically important molecules.
format	article
author	Aikaterini I. Argyriou Garyfallia I. Makrynitsa Georgios Dalkas Dimitra A. Georgopoulou Konstantinos Salagiannis Vassiliki Vazoura Andreas Papapetropoulos Stavros Topouzis Georgios A. Spyroulias
author_facet	Aikaterini I. Argyriou Garyfallia I. Makrynitsa Georgios Dalkas Dimitra A. Georgopoulou Konstantinos Salagiannis Vassiliki Vazoura Andreas Papapetropoulos Stavros Topouzis Georgios A. Spyroulias
author_sort	Aikaterini I. Argyriou
title	Replacement of heme by soluble guanylate cyclase (sGC) activators abolishes heme-nitric oxide/oxygen (H-NOX) domain structural plasticity
title_short	Replacement of heme by soluble guanylate cyclase (sGC) activators abolishes heme-nitric oxide/oxygen (H-NOX) domain structural plasticity
title_full	Replacement of heme by soluble guanylate cyclase (sGC) activators abolishes heme-nitric oxide/oxygen (H-NOX) domain structural plasticity
title_fullStr	Replacement of heme by soluble guanylate cyclase (sGC) activators abolishes heme-nitric oxide/oxygen (H-NOX) domain structural plasticity
title_full_unstemmed	Replacement of heme by soluble guanylate cyclase (sGC) activators abolishes heme-nitric oxide/oxygen (H-NOX) domain structural plasticity
title_sort	replacement of heme by soluble guanylate cyclase (sgc) activators abolishes heme-nitric oxide/oxygen (h-nox) domain structural plasticity
publisher	Elsevier
publishDate	2021
url	https://doaj.org/article/cace431cf9e043178301ce8c6e55771f
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Replacement of heme by soluble guanylate cyclase (sGC) activators abolishes heme-nitric oxide/oxygen (H-NOX) domain structural plasticity

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