Isoforms of Cyclic Nucleotide Phosphodiesterase PDE3 and Their Contribution to cAMP Hydrolytic Activity in Subcellular Fractions of Human Myocardium

Hambleton, Ryan; Krall, John M.; Tikishvili, Eliso; Honeggar, Matthew; Ahmad, Faiyaz; Manganiello, Vincent C.; Movsesian, Matthew A.

doi:10.1074/jbc.m506760200

Cited by 102 publications

(109 citation statements)

References 69 publications

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“…Growth of yeast on restricted media (A and B) and expression of a reporter construct, ␤-galactosidase (C and D), were used to assess the selectivity of the interaction between an amino-terminal fragment of HSPDE3B and 14-3-3 proteins (see "Experimental Procedures"). Yeast were transformed with either pEG202 and pACT2 (1 and 9); HSPDE3B (aa 2-90) and murine promyelocytic leukemic zinc finger protein (2 and 12); HSPDE3B (aa 2-90) and murine 14-3-3 (aa 70 -246) (3 and 14); HSPDE3B (aa 2-90) and murine 14-3-3 (aa 71-246) (4 and 13); HSPDE3B (aa 2-90) and pACT2 (5 and 11); pEG202 and PLZF (6); pEG202 and murine 14-3-3 (aa 70 -246) (7 and 10); pEG202 and murine 14-3-3 (aa 71-246) (8),; HSPDE3B (aa 2-90) and D. discoideum RVS161 homologue (15); and pEG202 and D. discoideum RVS161 homologue (16). E, PKA-dependent in vitro phosphorylation of a GST-HSPDE3B chimera encoding amino acids 2-90 of HSPDE3B.…”

Section: Resultsmentioning

confidence: 99%

“…Full-length PDE3A and PDE3B contain two N-terminal hydrophobic regions (NHR1 and NHR2) that target these enzymes to the endoplasmic reticulum and perhaps the plasma membrane (13)(14)(15)(16). Both PDE3A and PDE3B are substrates of protein kinase A (PKA) or protein kinase B (PKB), and activation of these kinases can result in phosphorylation-mediated activation of these enzymes in some cells (13)(14)(15)(16).…”

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confidence: 99%

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Protein Kinase A Phosphorylation of Human Phosphodiesterase 3B Promotes 14-3-3 Protein Binding and Inhibits Phosphatase-catalyzed Inactivation

Palmer

Jimmo

Raymond

et al. 2007

Journal of Biological Chemistry

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Recent studies confirm that intracellular cAMP concentrations are nonuniform and that localized subcellular cAMP hydrolysis by cyclic nucleotide phosphodiesterases (PDEs) is important in maintaining these cAMP compartments. Human phosphodiesterase 3B (HSPDE3B), a member of the PDE3 family of PDEs, represents the dominant particulate cAMP-PDE activity in many cell types, including adipocytes and cells of hematopoietic lineage. Although several previous reports have shown that phosphorylation of HSPDE3B by either protein kinase A (PKA) or protein kinase B (PKB) activates this enzyme, the mechanisms that allow cells to distinguish these two activated forms of HSPDE3B are unknown. Here we report that PKA phosphorylates HSPDE3B at several distinct sites (Ser-73, Ser-296, and Ser-318), and we show that phosphorylation of HSPDE3B at Ser-318 activates this PDE and stimulates its interaction with 14-3-3 proteins. In contrast, although PKB-catalyzed phosphorylation of HSPDE3B activates this enzyme, it does not promote 14-3-3 protein binding. Interestingly, we report that the PKA-phosphorylated, 14-3-3 protein-bound, form of HSPDE3B is protected from phosphatase-dependent dephosphorylation and inactivation. In contrast, PKA-phosphorylated HSPDE3B that is not bound to 14-3-3 proteins is readily dephosphorylated and inactivated. Our data are presented in the context that a selective interaction between PKAactivated HSPDE3B and 14-3-3 proteins represents a mechanism by which cells can protect this enzyme from deactivation. Moreover, we propose that this mechanism may allow cells to distinguish between PKA-and PKB-activated HSPDE3B.

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Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

Protein Kinase A Phosphorylation of Human Phosphodiesterase 3B Promotes 14-3-3 Protein Binding and Inhibits Phosphatase-catalyzed Inactivation

Palmer

Jimmo

Raymond

et al. 2007

Journal of Biological Chemistry

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“…In experiments in which we quantified the contribution of PDE3 isoforms to cyclic nucleotide hydrolytic activity in human myocardium, we found a large amount of Ca 2ϩ /calmodulinstimulated cAMP hydrolytic activity in soluble fractions of this tissue (14). Whether this activity was present in cardiac myocytes or in nonmuscle cells in the myocardium was unclear.…”

Section: Pde1mentioning

confidence: 95%

“…rtPDE3A1 (formerly designated rtPDE3A-136) was expressed in Sf9 cells and prepared as previously described (14). Other rtPDE constructs were expressed either in yeast, using the expression vector YepC-PADH2d, or in Sf9 cells, using a baculovirus expression system.…”

Section: Methodsmentioning

confidence: 99%

Cyclic Nucleotide Phosphodiesterase PDE1C1 in Human Cardiac Myocytes

Vandeput

Wolda²,

Krall

et al. 2007

Journal of Biological Chemistry

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101

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Isoforms in the PDE1 family of cyclic nucleotide phosphodiesterases were recently found to comprise a significant portion of the cGMP-inhibited cAMP hydrolytic activity in human hearts. We examined the expression of PDE1 isoforms in human myocardium, characterized their catalytic activity, and quantified their contribution to cAMP hydrolytic and cGMP hydrolytic activity in subcellular fractions of this tissue. Western blotting with isoform-selective anti-PDE1 monoclonal antibodies showed PDE1C1 to be the principal isoform expressed in human myocardium. Immunohistochemical analysis showed that PDE1C1 is distributed along the Z-lines and M-lines of cardiac myocytes in a striated pattern that differs from that of the other major dual-specificity cyclic nucleotide phosphodiesterase in human myocardium, PDE3A. Most of the PDE1C1 activity was recovered in soluble fractions of human myocardium. It binds both cAMP and cGMP with K m values of ϳ1 M and hydrolyzes both substrates with similar catalytic rates. PDE1C1 activity in subcellular fractions was quantified using a new PDE1-selective inhibitor, IC295. At substrate concentrations of 0.1 M, PDE1C1 constitutes the great majority of cAMP hydrolytic and cGMP hydrolytic activity in soluble fractions and the majority of cGMP hydrolytic activity in microsomal fractions, whereas PDE3 constitutes the majority of cAMP hydrolytic activity in microsomal fractions. These results indicate that PDE1C1 is expressed at high levels in human cardiac myocytes with an intracellular distribution distinct from that of PDE3A and that it may have a role in the integration of cGMP-, cAMP-and Ca 2؉ -mediated signaling in these cells. PDE13 cyclic nucleotide phosphodiesterases are dual-specificity enzymes that bind and hydrolyze cAMP and cGMP in a mutually competitive manner (for review, see Ref. 1). Their activity is increased by their binding to Ca 2ϩ and calmodulin, a feature unique to the PDE1 family of cyclic nucleotide phosphodiesterases. Three PDE1 genes, PDE1A, PDE1B, and PDE1C, have been identified, and several protein isoforms are generated from these genes by alternative splicing. PDE1A and PDE1B isoforms have significantly higher affinities for cGMP than for cAMP, whereas PDE1C isoforms have similar affinities for both substrates (2-4). The modulation of the catalytic activity of PDE1 isoforms by Ca 2ϩ -dependent stimulation and their susceptibility to mutually competitive inhibition by cAMP and cGMP suggest that these enzymes may be points of interaction among several signaling pathways.Cyclic nucleotide phosphodiesterases are particularly important in cardiac muscle. In humans, alterations in cAMP metabolism and cAMP-mediated signaling leading to decreases in intracellular cAMP content and in the phosphorylation of some substrates of cAMP-dependent protein kinase are prominent features of the pathophysiology of heart failure, and inhibition of PDE3 cAMP hydrolytic activity has a major role in its treatment (5). Changes in cGMP-mediated signaling in cardiac disease have not been ch...

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“…Disruption of the cAMP‐related signaling could be induced, for example, by dysregulation of the cGMP‐inhibited phosphodiesterase 3, which preferentially hydrolyzes cAMP 13, 35. As such, disruption of the NO‐cGMP pathway would impair PKA activation through increased cAMP hydrolysis by phosphodiesterase 3.…”

Section: Discussionmentioning

confidence: 99%

Newly Identified NO‐Sensor Guanylyl Cyclase/Connexin 43 Association Is Involved in Cardiac Electrical Function

Crassous

Shu

Huang

et al. 2017

JAHA

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BackgroundGuanylyl cyclase, a heme‐containing α1β1 heterodimer (GC1), produces cGMP in response to Nitric oxide (NO) stimulation. The NO‐GC1‐cGMP pathway negatively regulates cardiomyocyte contractility and protects against cardiac hypertrophy–related remodeling. We recently reported that the β1 subunit of GC1 is detected at the intercalated disc with connexin 43 (Cx43). Cx43 forms gap junctions (GJs) at the intercalated disc that are responsible for electrical propagation. We sought to determine whether there is a functional association between GC1 and Cx43 and its role in cardiac homeostasis.Methods and Results GC1 and Cx43 immunostaining at the intercalated disc and coimmunoprecipitation from membrane fraction indicate that GC1 and Cx43 are associated. Mice lacking the α subunit of GC1 (GCα1 knockout mice) displayed a significant decrease in GJ function (dye‐spread assay) and Cx43 membrane lateralization. In a cardiac‐hypertrophic model, angiotensin II treatment disrupted the GC1‐Cx43 association and induced significant Cx43 membrane lateralization, which was exacerbated in GCα1 knockout mice. Cx43 lateralization correlated with decreased Cx43‐containing GJs at the intercalated disc, predictors of electrical dysfunction. Accordingly, an ECG revealed that angiotensin II–treated GCα1 knockout mice had impaired ventricular electrical propagation. The phosphorylation level of Cx43 at serine 365, a protein‐kinase A upregulated site involved in trafficking/assembly of GJs, was decreased in these models.Conclusions GC1 modulates ventricular Cx43 location, hence GJ function, and partially protects from electrical dysfunction in an angiotensin II hypertrophy model. Disruption of the NO‐cGMP pathway is associated with cardiac electrical disturbance and abnormal Cx43 phosphorylation. This previously unknown NO/Cx43 signaling could be a protective mechanism against stress‐induced arrhythmia.

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Isoforms of Cyclic Nucleotide Phosphodiesterase PDE3 and Their Contribution to cAMP Hydrolytic Activity in Subcellular Fractions of Human Myocardium

Cited by 102 publications

References 69 publications

Protein Kinase A Phosphorylation of Human Phosphodiesterase 3B Promotes 14-3-3 Protein Binding and Inhibits Phosphatase-catalyzed Inactivation

Protein Kinase A Phosphorylation of Human Phosphodiesterase 3B Promotes 14-3-3 Protein Binding and Inhibits Phosphatase-catalyzed Inactivation

Cyclic Nucleotide Phosphodiesterase PDE1C1 in Human Cardiac Myocytes

Newly Identified NO‐Sensor Guanylyl Cyclase/Connexin 43 Association Is Involved in Cardiac Electrical Function

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