Skip to main content
NCBI home page
Biochemical Journal logoLink to Biochemical Journal
. 1992 Apr 15;283(Pt 2):487–491. doi: 10.1042/bj2830487

The catalytic subunit of protein kinase A triggers activation of the type V cyclic GMP-specific phosphodiesterase from guinea-pig lung.

F Burns 1, I W Rodger 1, N J Pyne 1
PMCID: PMC1131061  PMID: 1315515

Abstract

The type V cyclic GMP phosphodiesterase was partially purified from the high-speed supernatant of guinea-pig lung. The isoenzyme displayed linear kinetics for cyclic GMP hydrolysis, with Km = 2.2 +/- 0.2 microM and Vmax. = 1.2 +/- 0.08 nmol/min per mg. The selective type V phosphodiesterase inhibitor Zaprinast inhibited cyclic GMP hydrolysis with IC50 (concn. giving 50% inhibition) = 0.45 +/- 0.08 microM. Isobutylmethylxanthine promoted a 3-fold increase in the binding of cyclic GMP to the isoenzyme. The addition of the catalytic subunit of protein kinase A to an activation cocktail containing the partially purified type V phosphodiesterase resulted in a marked increase in Vmax. for cyclic GMP hydrolysis (approximately 10-fold at 40 units of protein kinase A). We have suggested that protein kinase A triggers phosphorylation of the phosphodiesterase, which results in activation of phosphodiesterase activity. In addition, the sensitivity to inhibition by Zaprinast is severely decreased (the IC50 for inhibition is 7.5 +/- 1.1 microM), suggesting that the potency of phosphodiesterase inhibitors is effected by phosphorylation of the enzyme.

Full text

PDF
487

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Beavo J. A., Reifsnyder D. H. Primary sequence of cyclic nucleotide phosphodiesterase isozymes and the design of selective inhibitors. Trends Pharmacol Sci. 1990 Apr;11(4):150–155. doi: 10.1016/0165-6147(90)90066-H. [DOI] [PubMed] [Google Scholar]
  2. Francis S. H., Corbin J. D. Purification of cGMP-binding protein phosphodiesterase from rat lung. Methods Enzymol. 1988;159:722–729. doi: 10.1016/0076-6879(88)59067-5. [DOI] [PubMed] [Google Scholar]
  3. Harris A. L., Connell M. J., Ferguson E. W., Wallace A. M., Gordon R. J., Pagani E. D., Silver P. J. Role of low Km cyclic AMP phosphodiesterase inhibition in tracheal relaxation and bronchodilation in the guinea pig. J Pharmacol Exp Ther. 1989 Oct;251(1):199–206. [PubMed] [Google Scholar]
  4. Harrison S. A., Reifsnyder D. H., Gallis B., Cadd G. G., Beavo J. A. Isolation and characterization of bovine cardiac muscle cGMP-inhibited phosphodiesterase: a receptor for new cardiotonic drugs. Mol Pharmacol. 1986 May;29(5):506–514. [PubMed] [Google Scholar]
  5. Heyworth C. M., Wallace A. V., Houslay M. D. Insulin and glucagon regulate the activation of two distinct membrane-bound cyclic AMP phosphodiesterases in hepatocytes. Biochem J. 1983 Jul 15;214(1):99–110. doi: 10.1042/bj2140099. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Kilgour E., Anderson N. G., Houslay M. D. Activation and phosphorylation of the 'dense-vesicle' high-affinity cyclic AMP phosphodiesterase by cyclic AMP-dependent protein kinase. Biochem J. 1989 May 15;260(1):27–36. doi: 10.1042/bj2600027. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Loten E. G., Assimacopoulos-Jeannet F. D., Exton J. H., Park C. R. Stimulation of a low Km phosphodiesterase from liver by insulin and glucagon. J Biol Chem. 1978 Feb 10;253(3):746–757. [PubMed] [Google Scholar]
  8. Macphee C. H., Reifsnyder D. H., Moore T. A., Lerea K. M., Beavo J. A. Phosphorylation results in activation of a cAMP phosphodiesterase in human platelets. J Biol Chem. 1988 Jul 25;263(21):10353–10358. [PubMed] [Google Scholar]
  9. Makino H., Kono T. Characterization of insulin-sensitive phosphodiesterase in fat cells. II. Comparison of enzyme activities stimulated by insulin and by isoproterenol. J Biol Chem. 1980 Aug 25;255(16):7850–7854. [PubMed] [Google Scholar]
  10. Nakatsu K., Diamond J. Role of cGMP in relaxation of vascular and other smooth muscle. Can J Physiol Pharmacol. 1989 Apr;67(4):251–262. doi: 10.1139/y89-042. [DOI] [PubMed] [Google Scholar]
  11. Pyne N. J., Cushley W., Nimmo H. G., Houslay M. D. Insulin stimulates the tyrosyl phosphorylation and activation of the 52 kDa peripheral plasma-membrane cyclic AMP phosphodiesterase in intact hepatocytes. Biochem J. 1989 Aug 1;261(3):897–904. doi: 10.1042/bj2610897. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Sharma R. K., Wang J. H. Calmodulin and Ca2+-dependent phosphorylation and dephosphorylation of 63-kDa subunit-containing bovine brain calmodulin-stimulated cyclic nucleotide phosphodiesterase isozyme. J Biol Chem. 1986 Jan 25;261(3):1322–1328. [PubMed] [Google Scholar]
  13. Souness J. E., Brazdil R., Diocee B. K., Jordan R. Role of selective cyclic GMP phosphodiesterase inhibition in the myorelaxant actions of M&B 22,948, MY-5445, vinpocetine and 1-methyl-3-isobutyl-8-(methylamino)xanthine. Br J Pharmacol. 1989 Nov;98(3):725–734. doi: 10.1111/j.1476-5381.1989.tb14599.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Thomas M. K., Francis S. H., Corbin J. D. Characterization of a purified bovine lung cGMP-binding cGMP phosphodiesterase. J Biol Chem. 1990 Sep 5;265(25):14964–14970. [PubMed] [Google Scholar]
  15. Thomas M. K., Francis S. H., Corbin J. D. Substrate- and kinase-directed regulation of phosphorylation of a cGMP-binding phosphodiesterase by cGMP. J Biol Chem. 1990 Sep 5;265(25):14971–14978. [PubMed] [Google Scholar]
  16. Thompson W. J., Appleman M. M. Multiple cyclic nucleotide phosphodiesterase activities from rat brain. Biochemistry. 1971 Jan 19;10(2):311–316. [PubMed] [Google Scholar]
  17. Torphy T. J., Burman M., Huang L. B., Tucker S. S. Inhibition of the low km cyclic AMP phosphodiesterase in intact canine trachealis by SK&F 94836: mechanical and biochemical responses. J Pharmacol Exp Ther. 1988 Sep;246(3):843–850. [PubMed] [Google Scholar]
  18. Zinman B., Hollenberg C. H. Effect of insulin and lipolytic agents on rat adipocyte low Km cyclic adenosine 3':5'-monophosphate phosphodiesterase. J Biol Chem. 1974 Apr 10;249(7):2182–2187. [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES