Evidence for phosphorylation of rat brain guanylate cyclase by cyclic AMP-dependent protein kinase

doi:10.1016/0006-291X(81)91600-4

Biochemical and Biophysical Research Communications

Volume 101, Issue 4, 31 August 1981, Pages 1381-1387

https://doi.org/10.1016/0006-291X(81)91600-4 Get rights and content

Abstract

Direct phosphorylation of purified rat brain guanylate cyclase by cyclic AMP-dependent protein kinase is demonstrated. In the presence of [γ-³²P]ATP, ³²P was incorporated into the protein to the extent of 0.8 to 0.9 mol/mol of guanylate cyclase. The presence of ³²P in the guanylate cyclase molecule was demonstrated by gel-filtration and by autoradiography after gel electrophoresis. The phosphorylation was accompanied by an increase in enzyme activity, characterized by an increase of V_M. These results suggest that the activity of guanylate cyclase may be regulated in vivo by phosphorylation.

References (14)

J. Zwiller et al.
Biochim. Biophys. Acta
(1981)
O.H. Lowry et al.
J. Biol. Chem
(1951)
E.M. Reimann et al.
J. Biol. Chem
(1971)
D.L. Garbers
J. Biol. Chem
(1979)
M. Weller
Biochim. Biophys. Acta
(1978)
T. Yamauchi et al.
Biochem. Biophys. Res. Commun
(1978)
J.K. Huttunen et al.
Biochim. Biophys. Acta
(1971)

There are more references available in the full text version of this article.

Cited by (58)

Involvement of the nitric oxide cascade in melatonin-induced inhibition of long-term potentiation at hippocampal CA1 synapses
2011, Neuroscience Research
Citation Excerpt :
Second, since PKG (cGKII) activated by cGMP is involved in GluR1 trafficking systems (Serulle et al., 2007) and since PKA phosphorylates GluR1 subunits (Esteban et al., 2003), these signaling pathways might interact via the inhibition of GluR1 trafficking. Third, since cGMP can be synthesized by GC phosphorylated by PKA activation (Zwiller et al., 1981), experimental manipulation of PKA may result in a change in the cGMP level. Also, in our present study, pre-incubation with the PKA inhibitor H-89 did not additively inhibit the degree of LTP in comparison with that produced by the application of melatonin and l-NAME without H-89 pre-incubation (Fig. 6), suggesting an interaction between the NO-cGMP and AC-PKA pathways.
Hippocampal long-term potentiation (LTP) is reportedly reduced in the presence of melatonin, but the cellular mechanisms of LTP inhibition by melatonin remain unclear. Since melatonin has the ability to scavenge free radicals such as nitric oxide (NO) and since NO has been suggested as an important contributor to LTP induction, in the present study we electrophysiologically examined whether melatonin inhibits hippocampal LTP by way of the NO signaling pathway. Field EPSP at Schaffer collateral – CA1 pyramidal cell synapses were recorded, and LTP was induced by tetanic stimulation (100 Hz, 1 s). Melatonin (100 nM) reduced the degree of LTP, and l-NAME (100 μM), an inhibitor of NO synthase, also reduced LTP, but simultaneous application of melatonin and l-NAME did not evoke any additional reduction of LTP in comparison with when only melatonin or only l-NAME were applied. Furthermore, the inhibition of LTP by the application of melatonin and l-NAME was disrupted by the application of an NO donor, DEA/NO (3 μM). The paired-pulse facilitation ratios before and after LTP induction by tetanic stimulation were nearly identical in the absence and presence of l-NAME. These results demonstrate that the inhibition of LTP in the presence of melatonin is due to the action of melatonin on the postsynaptic NO signaling pathway.
Soluble Guanylate Cyclase. Allosteric Activation and Redox Regulation.
2010, Nitric Oxide
This chapter focuses on a recently discovered family of allosteric enzyme activators, the soluble guanylate cyclase (sGC) stimulators and sGC activators, and their pharmacology, mechanism of enzyme activation, and therapeutic potential. It summarizes the evidence intimating that these new classes of sGC agonists have given rise to the hypothesis that sGC is an exquisite sensor of changes in cellular redox status and can adapt accordingly to preserve cytoprotective cGMP-dependent signaling. sGC is a principal receptor for the ubiquitous signaling molecule, nitric oxide (NO), and thereby plays a pivotal role in regulating cellular function in most organ systems. This heterodimeric, heme-containing protein catalyses the formation of the second messenger cyclic guanosine-3’,5’-monophosphate (cGMP), which in turn activates specific downstream effectors including kinases, phosphodiesterases, and ion channels. Novel allosteric activators of the protein stimulate cGMP formation, both its heme-containing as well as heme-free state (so-called sGC activators and sGC stimulators, respectively). This new family of sGC agonists provides the rationale for the thesis that sGC bioactivity is redox regulated and that this may be key to the pathogenesis of several cardiovascular disorders; moreover, this offers the opportunity to target drugs to diseased blood vessels or myocardium. This is exemplified by the current phase II/III clinical trials with the NO- and heme-independent sGC activator, Cinaciguat, for the intravenous treatment of acute decompensated heart failure and the NO-independent and heme-dependent sGC stimulator, riociguat, as an oral therapy for pulmonary hypertension.
Soluble Guanylate Cyclase
2009, Nitric Oxide: Biology and Pathobiology
This chapter focuses on a recently discovered family of allosteric enzyme activators, the soluble guanylate cyclase (sGC) stimulators and sGC activators, and their pharmacology, mechanism of enzyme activation, and therapeutic potential. It summarizes the evidence intimating that these new classes of sGC agonists have given rise to the hypothesis that sGC is an exquisite sensor of changes in cellular redox status and can adapt accordingly to preserve cytoprotective cGMP-dependent signaling. sGC is a principal receptor for the ubiquitous signaling molecule, nitric oxide (NO), and thereby plays a pivotal role in regulating cellular function in most organ systems. This heterodimeric, heme-containing protein catalyses the formation of the second messenger cyclic guanosine-3’,5’-monophosphate (cGMP), which in turn activates specific downstream effectors including kinases, phosphodiesterases, and ion channels. Novel allosteric activators of the protein stimulate cGMP formation, both its heme-containing as well as heme-free state (so-called sGC activators and sGC stimulators, respectively). This new family of sGC agonists provides the rationale for the thesis that sGC bioactivity is redox regulated and that this may be key to the pathogenesis of several cardiovascular disorders; moreover, this offers the opportunity to target drugs to diseased blood vessels or myocardium. This is exemplified by the current phase II/III clinical trials with the NO- and heme-independent sGC activator, Cinaciguat, for the intravenous treatment of acute decompensated heart failure and the NO-independent and heme-dependent sGC stimulator, riociguat, as an oral therapy for pulmonary hypertension.
Reactive oxygen species induce tyrosine phosphorylation of and Src kinase recruitment to NO-sensitive guanylyl cyclase
2005, Journal of Biological Chemistry
Citation Excerpt :
Indeed, it has been suggested that modifications such as reversible phosphorylation could add another tier of complexity to sGC regulation. For instance some reports have documented the phosphorylation of sGC on Ser/Thr residues (19–24). Also, Tyr phosphorylation has been implicated in sGC regulation (25); however, the molecular details underlying these processes and the biological consequences of sGC phosphorylation are still obscure.
Soluble guanylyl cyclase (sGC) is the major cytosolic receptor for nitric oxide (NO) that converts GTP into the second messenger cGMP in a NO-dependent manner. Other factors controlling this key enzyme are intracellular proteins such as Hsp90 and PSD95, which bind to sGC and modulate its activity, stability, and localization. To date little is known about the effects of posttranslational modifications of sGC, although circumstantial evidence suggests that reversible phosphorylation may contribute to sGC regulation. Here we demonstrate that inhibitors of protein-tyrosine phosphatases such as pervanadate and bisperoxo(1,10-phenanthroline)oxovanadate(V) as well as reactive oxygen species such as H₂O₂ induce specific tyrosine phosphorylation of the β₁ but not of the α₁ subunit of sGC. Tyrosine phosphorylation of sGCβ₁ is also inducible by pervanadate and H₂O₂ in intact PC12 cells, rat aortic smooth muscle cells, and in rat aortic tissues, indicating that tyrosine phosphorylation of sGC may also occur in vivo. We have mapped the major tyrosine phosphorylation site to position 192 of β₁, where it forms part of a highly acidic phospho-acceptor site for Src-like kinases. In the phosphorylated state Tyr(P)-192 exposes a docking site for SH2 domains and efficiently recruits Src and Fyn to sGCβ₁, thereby promoting multiple phosphorylation of the enzyme. Our results demonstrate that sGC is subject to tyrosine phosphorylation and interaction with Src-like kinases, revealing an unexpected cross-talk between the NO/cGMP and tyrosine kinase signaling pathways at the level of sGC.
Neurons exposed to ammonia reproduce the differential alteration in nitric oxide modulation of guanylate cyclase in the cerebellum and cortex of patients with liver cirrhosis
2005, Neurobiology of Disease
The activation of soluble guanylate cyclase by nitric oxide is increased in the frontal cortex but is reduced in the cerebellum of patients who died with liver cirrhosis. The aims of this work were to assess whether hyperammonemia is responsible for the region-selective alterations in guanylate cyclase modulation in liver cirrhosis and to assess whether the alteration occurs in neurons or in astrocytes. The activation of guanylate cyclase by nitric oxide was lower in cerebellar neurons exposed to ammonia (1.5-fold) than in control neurons (3.3-fold). The activation of guanylate cyclase by nitric oxide was higher in cortical neurons exposed to ammonia (8.7-fold) than in control neurons (5.5-fold). The activation was not affected in cerebellar or cortical astrocytes. These findings indicate that hyperammonemia is responsible for the differential alterations in the modulation of soluble guanylate cyclase in cerebellum and cerebral cortex of cirrhotic patients. Moreover, the alterations occur specifically in neurons and not in astrocytes.
Soluble guanylyl cyclase: More secrets revealed
2005, Cellular Signalling
Guanylyl cyclases (GCs) are enzymes that convert guanosine-5′-triphosphate (GTP) to cyclic guanosine-3′,5′-monophosphate (cGMP). The second messenger cGMP participates in signaling by (1) stimulating the activity of kinases that belong to the protein kinase G family, (2) altering the conductance of cGMP-gated ion channels and (3) changing the activity of cGMP-regulated phosphodiesterases. In contrast to adenylyl cyclases which exist as membrane-bound molecules, guanylyl cyclases (GC) occur in both membrane-bound and cytosolic forms. The particulate GC (pGC) isoforms serve as receptors for natriuretic peptides, while soluble GC (sGC) is the “receptor” for nitric oxide (NO). In addition to the difference in ligands and subcellular organization, the two forms of GC also differ in that pGC exists in homodimeric form, while typically sGC occurs as a heterodimer. Herein, we will review the literature on sGC subunit structure and discuss the regulation of the enzyme at the transcriptional and post-translational level.

View all citing articles on Scopus

View full text

Evidence for phosphorylation of rat brain guanylate cyclase by cyclic AMP-dependent protein kinase

Abstract

Biochim. Biophys. Acta

J. Biol. Chem

J. Biol. Chem

J. Biol. Chem

Biochim. Biophys. Acta

Biochem. Biophys. Res. Commun

Biochim. Biophys. Acta