Activation of soluble guanylate cyclase by NO-hemoproteins involves NO-heme exchange. Comparison of heme-containing and heme-deficient enzyme forms.

Ignarro, Louis J.; Adams, J.B.; Horwitz, P. M.; Wood, Keith S.

doi:10.1016/s0021-9258(19)89205-0

Cited by 240 publications

(39 citation statements)

References 17 publications

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“…Indeed, our work establishes how NO can operate in cells and tissues by controlling heme binding in proteins. This role is consistent with NO causing heme remobilization into the cytosol ( 50 ) and its capacity to promote heme transfer between purified proteins ( 51 ).…”

Section: Discussionsupporting

confidence: 70%

“…Because the ferrous heme–NO complex of sGCβ is highly stable ( 14 , 59 ), such NO–heme binding would create a positive thermodynamic driving force that supports a heme–NO species transfer from GAPDH to apo-sGCβ. Indeed, NO can promote a heme transfer from purified myoglobin to apo-sGC ( 51 , 60 ), and how thermodynamic gradients might govern heme transfers within cells has been discussed ( 61 ). Our current study shows that heme transfer between proteins can quickly take place in living cells in response to low concentrations of NO, indicating that the process is biologically relevant.…”

Section: Discussionmentioning

confidence: 99%

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NO rapidly mobilizes cellular heme to trigger assembly of its own receptor

Dai

Faul

Ghosh

et al. 2022

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

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Nitric oxide (NO) signaling in biology relies on its activating cyclic guanosine monophosphate (cGMP) production by the NO receptor soluble guanylyl cyclase (sGC). sGC must obtain heme and form a heterodimer to become functional, but paradoxically often exists as an immature heme-free form in cells and tissues. Based on our previous finding that NO can drive sGC maturation, we investigated its basis by utilizing a fluorescent sGC construct whose heme level can be monitored in living cells. We found that NO generated at physiologic levels quickly triggered cells to mobilize heme to immature sGC. This occurred when NO was generated within cells or by neighboring cells, began within seconds of NO exposure, and led cells to construct sGC heterodimers and thus increase their active sGC level by several-fold. The NO-triggered heme deployment involved cellular glyceraldehyde-3-phosphate dehydrogenase (GAPDH)–heme complexes and required the chaperone hsp90, and the newly formed sGC heterodimers remained functional long after NO generation had ceased. We conclude that NO at physiologic levels triggers assembly of its own receptor by causing a rapid deployment of cellular heme. Redirecting cellular heme in response to NO is a way for cells and tissues to modulate their cGMP signaling and to more generally tune their hemeprotein activities wherever NO biosynthesis takes place.

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Section: Discussionsupporting

confidence: 70%

Section: Discussionmentioning

confidence: 99%

NO rapidly mobilizes cellular heme to trigger assembly of its own receptor

Dai

Faul

Ghosh

et al. 2022

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

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“…Our past [20] and present [53] findings also revealed that low doses of nitric oxide (NO) can contribute to sGC maturation by triggering a rapid Hsp90-dependent heme insertion into the apo-sGCβ1 (heme-free) population, ultimately resulting in a mature sGC-α1β1 heterodimer [20]. This finding of an elevated active sGC-α1β1 heterodimer formation by a NO trigger causing heme-insertion into apo-sGCβ1, filled a void in an earlier work done by Ignarro and colleagues in the 1980s which showed that NO-heme moiety could be transferred into heme-free or apo-sGCβ1 through an exchange reaction with NO-hemeproteins to activate the enzyme [54]. This suggests that a NO-Hsp90 synergy may be essential for maturation and activation of certain hemeproteins.…”

Section: Role Of Hsp90 In Client Hemeprotein Maturationmentioning

confidence: 89%

Hsp90 in Human Diseases: Molecular Mechanisms to Therapeutic Approaches

Sumi

Ghosh

2022

Cells

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The maturation of hemeprotein dictates that they incorporate heme and become active, but knowledge of this essential cellular process remains incomplete. Studies on chaperon Hsp90 has revealed that it drives functional heme maturation of inducible nitric oxide synthase (iNOS), soluble guanylate cyclase (sGC) hemoglobin (Hb) and myoglobin (Mb) along with other proteins including GAPDH, while globin heme maturations also need an active sGC. In all these cases, Hsp90 interacts with the heme-free or apo-protein and then drives the heme maturation by an ATP dependent process before dissociating from the heme-replete proteins, suggesting that it is a key player in such heme-insertion processes. As the studies on globin maturation also need an active sGC, it connects the globin maturation to the NO-sGC (Nitric oxide-sGC) signal pathway, thereby constituting a novel NO-sGC-Globin axis. Since many aggressive cancer cells make Hbβ/Mb to survive, the dependence of the globin maturation of cancer cells places the NO-sGC signal pathway in a new light for therapeutic intervention. Given the diverse role of chaperon Hsp90, and in particular to it being a major therapeutic target in drug discovery programs, the current findings open up more avenues of therapeutic intervention where these hemeproteins are either overactive or are dysfunctional.

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“…It is possible that other cell proteins or small molecules may alter the heme affinity of GAPDH or its protein interactions to promote or inhibit heme delivery. One factor that may be involved is NO, which can modify GAPDH function through S-nitrosation of its Cys residues (35,36) and can positively or negatively impact heme delivery to sGC (16,37) and to several other heme proteins (38,39). Finally, which proteins undergo GAPDH-dependent versus GAPDH-independent heme delivery?…”

Section: Discussionmentioning

confidence: 99%

GAPDH delivers heme to soluble guanylyl cyclase

Dai

Sweeny

Schlanger

et al. 2020

Journal of Biological Chemistry

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Soluble guanylyl cyclase (sGC) is a key component of NO–cGMP signaling in mammals. Although heme must bind in the sGC β1 subunit (sGCβ) for sGC to function, how heme is delivered to sGCβ remains unknown. Given that GAPDH displays properties of a heme chaperone for inducible NO synthase, here we investigated whether heme delivery to apo-sGCβ involves GAPDH. We utilized an sGCβ reporter construct, tetra-Cys sGCβ, whose heme insertion can be followed by fluorescence quenching in live cells, assessed how lowering cell GAPDH expression impacts heme delivery, and examined whether expressing WT GAPDH or a GAPDH variant defective in heme binding recovers heme delivery. We also studied interaction between GAPDH and sGCβ in cells and their complex formation and potential heme transfer using purified proteins. We found that heme delivery to apo-sGCβ correlates with cellular GAPDH expression levels and depends on the ability of GAPDH to bind intracellular heme, that apo-sGCβ associates with GAPDH in cells and dissociates when heme binds sGCβ, and that the purified GAPDH–heme complex binds to apo-sGCβ and transfers its heme to sGCβ. On the basis of these results, we propose a model where GAPDH obtains mitochondrial heme and then forms a complex with apo-sGCβ to accomplish heme delivery to sGCβ. Our findings illuminate a critical step in sGC maturation and uncover an additional mechanism that regulates its activity in health and disease.

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Activation of soluble guanylate cyclase by NO-hemoproteins involves NO-heme exchange. Comparison of heme-containing and heme-deficient enzyme forms.

Cited by 240 publications

References 17 publications

NO rapidly mobilizes cellular heme to trigger assembly of its own receptor

NO rapidly mobilizes cellular heme to trigger assembly of its own receptor

Hsp90 in Human Diseases: Molecular Mechanisms to Therapeutic Approaches

GAPDH delivers heme to soluble guanylyl cyclase

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