Nitric oxide and heat shock protein 90 activate soluble guanylate cyclase by driving rapid change in its subunit interactions and heme content

Ghosh, Arnab; Stasch, Johannes‐Peter; Papapetropoulos, Andreas; Stuehr, Dennis J.

doi:10.1016/j.niox.2014.09.044

Cited by 8 publications

(29 citation statements)

References 15 publications

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“…We found this change in sGC-β1 protein association occurred in lung samples from the asthmatic mice, and also in human lung slices or HASMC that had been exposed to either chemical-or cell-derived NO in a manner that mimicked the chronic airway NO exposure that is seen in asthma. Our previous work (15) showed that BAY 60-2770 could convert the inactive, hsp90-associated form of sGC-β1 back into active sGC-α1β1 heterodimers, and our current findings are consistent with this conversion likely being its mode of action for activating the NO-resistant sGC subpopulation that built up in the asthmatic airways. Regarding BAY 41-2272, we saw that it remained effective in causing bronchodilation in our asthmatic mouse models, but its relative efficacy is likely tied to the extent to which a mature sGC subpopulation remains intact in the inflammed lungs, which in turn may be inversely related to the degree of lung inflammation.…”

Section: Discussionsupporting

confidence: 89%

“…Five days later animals received five daily intranasal challenges of 10 μg HDME in 50 μL saline (32,33 Western Blots and Immunoprecipitations. Standard protocols were followed as previously mentioned (14,15). For immunoprecipitations (IP), 500 μg of the total mouse/human PCLS lung or RFL-6 cell supernatant was precleared with 20 μL of protein G-Sepharose beads (Amersham) for 1 h at 4°C, beads were pelleted, and the supernatants incubated overnight at 4°C with 3 μg of antisGC-β1 antibody.…”

Section: Methodsmentioning

confidence: 99%

“…In certain other cases, sGC activity in the cell supernatants was determined by first passing them through the desalting G-25 PD-spin trap columns. Enzymatic activity (15,34) was determined in 10-min incubations containing the protein solution, 500 μM GTP, 20 μM of sGC activators BAY 41-2272 or BAY 60-2770, and buffer containing 50 mM triethanolamine/HCl pH 7.4, 3 mM MgCl 2, 3 mM DTT, and 250 μM IBMX at 37°C. Reactions were quenched by addition of 10 mM Na 2 CO 3 and Zn (CH 3 CO 3 ) 2 .…”

Section: Methodsmentioning

confidence: 99%

“…2), and implies that the allergic inflammation caused a significant portion of the lung sGC to accumulate in an oxidatively damaged and NO-unresponsive-but BAY 60-2270-responsive-form. Indeed, the sGC-β1 from the asthmatic lungs exhibited three biochemical hallmarks of being damaged and NO-unresponsive (14,15): an increased level of cysteine S-nitrosylation ( Fig. 3 B and C), a diminished association with its partner sGC-α1 subunit, and an enhanced association with the cellular chaperone hsp90 (Fig.…”

Section: Biomarkers Of Sgc Damage Manifest In the Mouse Asthmatic Lungmentioning

confidence: 99%

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Soluble guanylate cyclase as an alternative target for bronchodilator therapy in asthma

Ghosh

Koziol‐White

Asosingh

et al. 2016

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

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119

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Asthma is defined by airway inflammation and hyperresponsiveness, and contributes to morbidity and mortality worldwide. Although bronchodilation is a cornerstone of treatment, current bronchodilators become ineffective with worsening asthma severity. We investigated an alternative pathway that involves activating the airway smooth muscle enzyme, soluble guanylate cyclase (sGC). Activating sGC by its natural stimulant nitric oxide (NO), or by pharmacologic sGC agonists BAY 41-2272 and BAY 60-2770, triggered bronchodilation in normal human lung slices and in mouse airways. Both BAY 41-2272 and BAY 60-2770 reversed airway hyperresponsiveness in mice with allergic asthma and restored normal lung function. The sGC from mouse asthmatic lungs displayed three hallmarks of oxidative damage that render it NO-insensitive, and identical changes to sGC occurred in human lung slices or in human airway smooth muscle cells when given chronic NO exposure to mimic the high NO in asthmatic lung. Our findings show how allergic inflammation in asthma may impede NO-based bronchodilation, and reveal that pharmacologic sGC agonists can achieve bronchodilation despite this loss.A sthma is an inflammatory disease that causes airway hyperreactivity (AHR) and bronchoconstriction, which impedes daily life activities and, when severe, can cause death. It is the most common chronic disease of childhood, accounts for one in three emergency department visits daily, and asthma diagnoses are increasing worldwide (1). The leading treatment for relief and acute care is bronchodilation, which relies heavily on the β-adrenergic receptor-cAMP pathway. Nearly 70% of patients, however, develop resistance or tachyphylaxis to the existing β-agonist therapy (2), underscoring a need for new bronchodilators that can act through a different pharmacologic principle.The nitric oxide-soluble guanylate cyclase-cGMP pathway (NO-sGC-cGMP) is the primary signal transduction pathway for relaxing vascular smooth muscle (3). In contrast, a role for the NO-sGC-cGMP pathway in relaxing airway smooth muscle is less clear (4, 5), and bronchodilation was instead suggested to depend on glutathione nitrosothiol levels in the lung (6, 7). However, recent studies have shown that inflammation can desensitize sGC toward its natural activator, NO (8), and new drugs have become available that directly activate sGC, independent of NO (9). These developments encouraged us to re-examine the NO-sGC-cGMP pathway regarding its role in bronchodilation, its becoming damaged in inflammatory asthma, and its potential for alternative bronchodilator development under this circumstance. ResultsThe NO-sGC-cGMP Pathway Bronchodilates Human Lung. We first tested if stimulating the NO-sGC-cGMP pathway would dilate preconstricted small airways in human precision-cut lung slices (PCLS) obtained from healthy donor lungs (Fig. 1A and Table S1). Graded doses of the slow-release NO donor DETA/NO [3,3-Bis(aminoethyl)-1-hydroxy-2-oxo-1-triazene] produced bronchodilation in human PCLS similar to what was ...

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Section: Discussionsupporting

confidence: 89%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Biomarkers Of Sgc Damage Manifest In the Mouse Asthmatic Lungmentioning

confidence: 99%

See 2 more Smart Citations

Soluble guanylate cyclase as an alternative target for bronchodilator therapy in asthma

Ghosh

Koziol‐White

Asosingh

et al. 2016

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

Self Cite

119

View full text Add to dashboard Cite

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“…HSP90-mediated heme incorporation converts apo-sGC into a mature active form that is responsive to NO [77]. The insertion of heme into sGC is positively regulated by NO [89]. HRG-1 and its mammalian homologs also transport heme within the cell.…”

Section: Intracellular Heme Transportmentioning

confidence: 99%

Regulation of heme biosynthesis and transport in metazoa

Sun

Cheng

Chen

2015

Sci. China Life Sci.

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Heme is an iron-containing tetrapyrrole that plays a critical role in regulating a variety of biological processes including oxygen and electron transport, gas sensing, signal transduction, biological clock, and microRNA processing. Most metazoan cells synthesize heme via a conserved pathway comprised of eight enzyme-catalyzed reactions. Heme can also be acquired from food or extracellular environment. Cellular heme homeostasis is maintained through the coordinated regulation of synthesis, transport, and degradation. This review presents the current knowledge of the synthesis and transport of heme in metazoans and highlights recent advances in the regulation of these pathways. heme, iron, synthesis, transport, regulation Citation:Sun FX, Cheng YJ, Chen CY. Regulation of heme biosynthesis and transport in metazoa.

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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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Nitric oxide and heat shock protein 90 activate soluble guanylate cyclase by driving rapid change in its subunit interactions and heme content

Cited by 8 publications

References 15 publications

Soluble guanylate cyclase as an alternative target for bronchodilator therapy in asthma

Soluble guanylate cyclase as an alternative target for bronchodilator therapy in asthma

Regulation of heme biosynthesis and transport in metazoa

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

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