Hydrolysis of cyclic nucleotides by a purified cGMP-stimulated phosphodiesterase: structural requirements for hydrolysis

Braumann, Thomas; Erneux, Christophé; Petridis, Georg; Stohrer, Wolf‐Dieter; Jastorff, Bernd

doi:10.1016/0167-4838(86)90174-3

Cited by 46 publications

(26 citation statements)

References 26 publications

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“…Because the magnitude and direction of the dipole moment of the purine ring in cGMP is significantly different than in cAMP, the guanine base would be expected to have different dipole-induced dipole interactions with the polarizable Phe-438 (40). These differences have been used to explain catalytic specificity preferences for cyclic nucleotide analogs at the PDE2 catalytic site (39,40). Actually, because these studies were conducted with PDE2 holoenzyme and we now know that the GAF domains can regulate the activity of the catalytic domain of the enzyme, it may be that the measured catalytic activities were in part due to differences in GAF domain binding of the analogs.…”

Section: Gaf-b Mutations Do Not Greatly Affect Global Stability Ormentioning

confidence: 99%

Molecular Determinants for Cyclic Nucleotide Binding to the Regulatory Domains of Phosphodiesterase 2A

Tang

Martinez

et al. 2004

Journal of Biological Chemistry

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Binding of cGMP to the GAF-B domain of phosphodiesterase 2A allosterically activates catalytic activity. We report here a series of mutagenesis studies on the GAF-B domain of PDE2A that support a novel mechanism for molecular recognition of cGMP. Alanine mutations of Phe-438, Asp-439, and Thr-488, amino acids that interact with the pyrimidine ring, decrease cGMP affinity slightly but increase cAMP affinity by up to 8-fold. Each interaction is required to provide for cAMP/cGMP specificity. Mutations of any of the residues that interact with the phosphate-ribose moiety or the imidazole ring abolish cGMP binding. Thus, residues that interact with the pyrimidine ring collectively control cAMP/cGMP specificity, whereas residues that bind the phosphateribose moiety and imidazole ring are critical for high affinity binding. Similar decreases in binding were found for mutations made in a bacterially expressed GAF-A/B plus catalytic domain construct. Because these constructs had very high catalytic activity, it appears that these mutations did not cause a global denaturation. The affinities of cAMP and cGMP for wild-type GAF-B alone were ϳ4-fold greater than for the holoenzyme, suggesting that the presence of neighboring domains alters the conformation of GAF-B. More importantly, the PDE2A GAF-B, GAF-A/B, GAF-A/B؉C domains, and holoenzyme all bind cGMP with much higher affinity than has previously been reported. This high affinity suggests that cGMP binding to PDE2 GAF-B activates the enzyme rapidly, stoichiometrically, and in an all or none fashion, rather than variably over a large range of cyclic nucleotide concentrations.

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Section: Gaf-b Mutations Do Not Greatly Affect Global Stability Ormentioning

confidence: 99%

Molecular Determinants for Cyclic Nucleotide Binding to the Regulatory Domains of Phosphodiesterase 2A

Tang

Martinez

et al. 2004

Journal of Biological Chemistry

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“…This effect can be mainly attributed to this transduction pathway since it was evoked by the Sp-phosphorothioate isomer of CAMP. This analog, which has a sulfur substitution in the exocyclic axial oxygen position, mimicks the natural second messenger CAMP, and is extremely stable against cyclic nucleotide phosphodiesterases (Braumann et al, 1986). So, undesired metabolites which could lead to biological effects as observed for dibutyryl-CAMP, 8ClcAMP, 8Br-CAMP, and N6-substituted cAMP analogs, cannot interfere (Burns et al, 1988;Van Lookeren Campagne et al, 1991;Martin and Kowalchyk, 1981).…”

Section: Discussionmentioning

confidence: 96%

Cyclic‐AMP inhibits cell growth and negatively interacts with platelet membrane glycoprotein expression on the Dami human megakaryoblastic cell line

Vittet¹,

Duperray²,

Chevillard³

1995

Journal Cellular Physiology

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Intracellular signaling processes by which hematopoietic growth factors regulate megakaryocytopoiesis remain incompletely understood. Cyclic AMP (cAMP) has been shown to be implicated in the regulation of growth and differentiation in various normal and malignant cell types. Since a few studies have suggested the possible involvement of the cAMP pathway as one of the intracellular mechanisms whereby megakaryocytopoiesis may be regulated, we investigated the functional effects of cAMP on the human megakaryoblastic Dami cell line. We observed that exposure of Dami cells to cAMP analogs or to agents elevating intracellular cAMP levels yielded dose-dependent cell growth inhibition. Cell cycle progression analysis of cells predominantly synchronized at the G1/S boundary by prior treatment with hydroxyurea revealed that cAMP transiently accumulated cells in the G2/M phase, then slowing down cell cycle. On the other hand, immunofluorescence and Northern blot analysis of megakaryocytic differentiation marker expression showed that probes we have used significantly inhibited GPIb expression. Moreover, although these agents used alone did not affect GPIIb/IIIa expression, they markedly reversed phorbol ester-induced GPIIb/IIIa expression increase. These inhibitory cAMP actions on glycoprotein expression were not the result of cell cycle perturbation since we observed that GPIb and GPIIb/IIIa expression were not cell cycle dependent. All these data may then be consistent with a potential negative regulatory role of the cAMP intracellular signaling pathway during megakaryocytopoiesis.

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“…Studies have shown however that a negative charge, an equatorial oxygen atom at the cyclic phosphate residue and the presence of divalent cations are absolute requirements for cleavage of the cyclic phosphodiester bond of cAMP or cGMP [23]. Goldberg et al [24] showed that PDE-catalyzed hydrolysis of cGMP occurs by nucleophilic substitution of a solvent hydroxyl (-OH) at the phosphate, which cleaves the P-O bond, opening the phosphodiester ring (see Fig.…”

Section: Overview Of Pde Biochemistrymentioning

confidence: 99%

Phosphodiesterase Type 5 Inhibitors: Molecular Pharmacology and Interactions with other Phosphodiesterases

Seftel¹

2005

CPD

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Erectile function is determined by tight regulation of relaxation or contraction of corpus cavernosal smooth muscle, which is the result of a long and complex chain of molecular events. Control of erectile function resides in signaling pathways of the central and peripheral nervous system, as well as intracellular events in the penile smooth muscle. Vascular events resulting in erection have long been understood, and the role of the signaling pathways of the central and peripheral nervous systems in erectile function and dysfunction has become increasingly clear over the last decade. This knowledge has led to the development and current availability of effective oral treatments for erectile dysfunction, the selective phosphodiesterase type 5 (PDE5) inhibitors-sildenafil, vardenafil and tadalafil. In the past few years we have seen an elucidation of the molecular events involved in erectile function and dysfunction and the detailed mechanisms of action by which the specific PDE5 inhibitors work. A review of those mechanisms helps to explain the success of the currently available PDE5 inhibitors and the differences between them and suggests new approaches for developing potential future novel therapies or refinements to existing structures that may improve their efficacy, selectivity and safety profiles.

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Hydrolysis of cyclic nucleotides by a purified cGMP-stimulated phosphodiesterase: structural requirements for hydrolysis

Cited by 46 publications

References 26 publications

Molecular Determinants for Cyclic Nucleotide Binding to the Regulatory Domains of Phosphodiesterase 2A

Molecular Determinants for Cyclic Nucleotide Binding to the Regulatory Domains of Phosphodiesterase 2A

Cyclic‐AMP inhibits cell growth and negatively interacts with platelet membrane glycoprotein expression on the Dami human megakaryoblastic cell line

Phosphodiesterase Type 5 Inhibitors: Molecular Pharmacology and Interactions with other Phosphodiesterases

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