Inhibition of cyclic GMP-dependent protein kinase-mediated effects by (Rp)-8-bromo-PET-cyclic GMPS
- PMID: 8719784
- PMCID: PMC1909162
- DOI: 10.1111/j.1476-5381.1995.tb15112.x
Inhibition of cyclic GMP-dependent protein kinase-mediated effects by (Rp)-8-bromo-PET-cyclic GMPS
Abstract
1. The modulation of the guanosine 3':5'-cyclic monophosphate (cyclic GMP)- and adenosine 3':5'-cyclic monophosphate (cyclic AMP)-dependent protein kinase activities by the diastereomers of 8-bromo-beta phenyl-1, N2-ethenoguanosine 3':5'-cyclic monophosphorothioate, ((Rp)- and (Sp)-8-bromo-PET-cyclic GMPS) was investigated by use of purified protein kinases. In addition, the effects of (Rp)-8-bromo-PET-cyclic GMPS on protein phosphorylation in intact human platelets and on [3H]-noradrenaline release and neurogenic vasoconstriction in electrical field stimulated rat tail arteries were also studied. 2. Kinetic analysis with purified cyclic GMP-dependent protein kinase (PKG) type I alpha and I beta, which are expressed in the rat tail artery, revealed that (Rp)-8-bromo-PET-cyclic GMPS is a competitive inhibitor with an apparent Ki of 0.03 microM. The activation of purified cyclic AMP-dependent protein kinase (PKA) type II was antagonized with an apparent Ki of 10 microM. 3. In human platelets, (Rp)-8-bromo-PET-cyclic GMPS (0.1 mM) antagonized the activation of the PKG by the selective activator 8-(4-chlorophenylthio)-guanosine 3':5'-cyclic monophosphate (8-pCPT-cyclic GMP; 0.2 mM) without affecting the activation of PKA by (Sp)-5, 6-dichloro-1-beta-D-ribofurano-sylbenzimidazole- 3':5'-cyclic monophosphorothioate ((Sp)-5,6-DCl-cyclic BiMPS; 0.1 mM). 4. (Rp)-8-bromo-PET-cyclic GMPS was not hydrolysed by the cyclic GMP specific phosphodiesterase (PDE) type V from bovine aorta but potently inhibited this PDE. 5. The corresponding sulphur free cyclic nucleotide of the two studied phosphorothioate derivatives, 8-bromo-beta-phenyl-1, N2-ethenoguanosine-3':5'-cyclic monophosphate (8-bromo-PET-cyclic GMP), had no effect on electrically-induced [3H]-noradrenaline release but concentration-dependently decreased the stimulation-induced vasoconstriction. (Rp)-8-bromo-PET-cyclic GMPS (3 microM) shifted the vasoconstriction response to the right without affecting stimulation evoked tritium overflow. 6. The NO donor, 3-morpholinosydnonimine (SIN-1) relaxed rat tail arteries precontracted with phenylephrine (1 microM). The SIN-1 concentration-relaxation curve was shifted in a parallel manner to the right by (Rp)-8-bromo-PET-cyclic GMPS, suggesting that the relaxation was mediated by a cyclic GMP/PKG-dependent mechanism. 7. The [3H]-noradrenaline release-enhancing effect and stimulation-induced decrease in vasoconstriction of forskolin were unaffected by (Rp)-8-bromo-PET-cyclic GMPS. Moreover, the forskolin concentration-relaxation curve was not changed in the presence of the PKG inhibitor, suggesting a high selectivity in intact cells for PKG- over PKA-mediated effects. 8. The results obtained indicate that (Rp)-8-bromo-PET-cyclic GMPS presently is the most potent and selective inhibitor of PKG and is helpful in distinguishing between cyclic GMP and cyclic AMP messenger pathways activation. Therefore, this phosphorothioate stereomer may be a useful tool for studying the role of cyclic GMP in vitro.
Similar articles
-
Exploring the mechanisms of vascular smooth muscle tone with highly specific, membrane-permeable inhibitors of cyclic GMP-dependent protein kinase Ialpha.Pharmacol Ther. 2002 Feb-Mar;93(2-3):203-15. doi: 10.1016/s0163-7258(02)00189-4. Pharmacol Ther. 2002. PMID: 12191612 Review.
-
Involvement of cyclic nucleotide-dependent protein kinases in cyclic AMP-mediated vasorelaxation.Br J Pharmacol. 1997 Sep;122(1):158-64. doi: 10.1038/sj.bjp.0701339. Br J Pharmacol. 1997. PMID: 9298542 Free PMC article.
-
Effects of cyclic AMP and analogues on neurogenic transmission in the rat tail artery.Br J Pharmacol. 1994 Feb;111(2):625-31. doi: 10.1111/j.1476-5381.1994.tb14782.x. Br J Pharmacol. 1994. PMID: 8004406 Free PMC article.
-
Effects of cyclic GMP and analogues on neurogenic transmission in the rat tail artery.Br J Pharmacol. 1994 Jul;112(3):867-72. doi: 10.1111/j.1476-5381.1994.tb13160.x. Br J Pharmacol. 1994. PMID: 7921614 Free PMC article.
-
Towards an understanding of the mechanism of action of cyclic AMP and cyclic GMP in smooth muscle relaxation.Blood Vessels. 1991;28(1-3):129-37. doi: 10.1159/000158852. Blood Vessels. 1991. PMID: 1848122 Review.
Cited by
-
Integrative Kinase Activity Profiling and Phosphoproteomics of rd10 Mouse Retina during cGMP-Dependent Retinal Degeneration.Int J Mol Sci. 2024 Mar 19;25(6):3446. doi: 10.3390/ijms25063446. Int J Mol Sci. 2024. PMID: 38542418 Free PMC article.
-
8-Br-cGMP suppresses tumor progression through EGFR/PLC γ1 pathway in epithelial ovarian cancer.Mol Biol Rep. 2024 Jan 18;51(1):140. doi: 10.1007/s11033-023-09037-5. Mol Biol Rep. 2024. PMID: 38236447
-
The PKG Inhibitor CN238 Affords Functional Protection of Photoreceptors and Ganglion Cells against Retinal Degeneration.Int J Mol Sci. 2023 Oct 17;24(20):15277. doi: 10.3390/ijms242015277. Int J Mol Sci. 2023. PMID: 37894958 Free PMC article.
-
The Phosphoproteome of the Rd1 Mouse Retina, a Model of Inherited Photoreceptor Degeneration, Changes after Protein Kinase G Inhibition.Int J Mol Sci. 2023 Jun 7;24(12):9836. doi: 10.3390/ijms24129836. Int J Mol Sci. 2023. PMID: 37372984 Free PMC article.
-
cGMP Analogues with Opposing Actions on CNG Channels Selectively Modulate Rod or Cone Photoreceptor Function.Pharmaceutics. 2022 Oct 1;14(10):2102. doi: 10.3390/pharmaceutics14102102. Pharmaceutics. 2022. PMID: 36297537 Free PMC article.
References
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Other Literature Sources
Molecular Biology Databases