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

F Burns, I W Rodger, N J Pyne

Research output: Contribution to journalArticle

73 Citations (Scopus)

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.
LanguageEnglish
Pages487-491
Number of pages5
JournalBiochemical journal
Volume283
Issue number2
Publication statusPublished - 15 Apr 1992

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Cyclic AMP-Dependent Protein Kinase Catalytic Subunits
Cyclic GMP
Phosphoric Diester Hydrolases
Guinea Pigs
Chemical activation
Lung
Hydrolysis
Phosphorylation
Phosphodiesterase Inhibitors
Cyclic AMP-Dependent Protein Kinases
Isoenzymes
Inhibitory Concentration 50
Type 5 Cyclic Nucleotide Phosphodiesterases
Kinetics
Enzymes

Keywords

  • 3',5'-cyclic-AMP phosphodiesterases
  • 3',5'-cyclic-GMP phosphodiesterases
  • animals
  • chromatography, affinity
  • chromatography, ion exchange
  • cytosolic
  • enzyme activation
  • guinea pigs
  • isoenzymes
  • kinetics
  • lung
  • macromolecular substances
  • protein kinases
  • purinones
  • substrate specificity

Cite this

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title = "The catalytic subunit of protein kinase A triggers activation of the type V cyclic GMP-specific phosphodiesterase from guinea-pig lung",
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.",
keywords = "3',5'-cyclic-AMP phosphodiesterases, 3',5'-cyclic-GMP phosphodiesterases, animals, chromatography, affinity, chromatography, ion exchange, cytosolic, enzyme activation, guinea pigs, isoenzymes, kinetics, lung, macromolecular substances, protein kinases, purinones, substrate specificity",
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The catalytic subunit of protein kinase A triggers activation of the type V cyclic GMP-specific phosphodiesterase from guinea-pig lung. / Burns, F; Rodger, I W; Pyne, N J.

In: Biochemical journal, Vol. 283 , No. 2, 15.04.1992, p. 487-491.

Research output: Contribution to journalArticle

TY - JOUR

T1 - The catalytic subunit of protein kinase A triggers activation of the type V cyclic GMP-specific phosphodiesterase from guinea-pig lung

AU - Burns, F

AU - Rodger, I W

AU - Pyne, N J

PY - 1992/4/15

Y1 - 1992/4/15

N2 - 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.

AB - 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.

KW - 3',5'-cyclic-AMP phosphodiesterases

KW - 3',5'-cyclic-GMP phosphodiesterases

KW - animals

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KW - chromatography, ion exchange

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KW - enzyme activation

KW - guinea pigs

KW - isoenzymes

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KW - lung

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KW - protein kinases

KW - purinones

KW - substrate specificity

M3 - Article

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SP - 487

EP - 491

JO - Biochemical Journal

T2 - Biochemical Journal

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