Cyclic nucleotide phosphodiesterases from frog atrial fibers: isolation and drug sensitivities.
The cyclic nucleotide phosphodiesterase (PDE) forms present in frog atrial fibers were isolated and characterized by their drug sensitivities. DEAE-sephacel chromatography of cytosolic PDE activity resolved three major PDE forms: peak A hydrolyzed both adenosine 3',5'-cyclic monophosphate (cAMP) and guanosine 3',5'-cyclic monophosphate (cGMP) and was activated by calcium-calmodulin (PDE I); peak B also hydrolyzed both cAMP and cGMP but was activated by 5 microM cGMP (PDE II); peak C specifically hydrolyzed cAMP (PDE IV). Rolipram specifically inhibited PDE IV (Ki = 1.1 microM), whereas dipyridamole potently inhibited both PDE II (Ki = 4.6 microM) and PDE IV (Ki = 0.8 microM). Atrial fiber PDE I was preferentially inhibited by zaprinast (Ki = 10 microM). 3-Isobutyl-1-methyl xanthine (IBMX) and theophylline inhibited nonspecifically all three different enzymes. The positive inotropic drug CI 930 only inhibited the different isolated atrial PDE forms at concentrations greater than 200 microM. However, under assay conditions for which PDE IV was specifically inhibited (presence of 100 microM rolipram), an IC50 of 17 microM for CI 930 was observed on the remaining 26% cAMP hydrolytic activity of peak C (which could represent a cGMP-inhibited PDE form: PDE III). The same PDE forms were also found in frog ventricle. The major difference between frog atrial fiber (and ventricular tissue) PDEs and mammalian cardiac PDEs is that the main cytosolic cAMP-specific hydrolytic activity in frog heart is due to PDE IV rather than PDE III. Rolipram, dipyridamole, and zaprinast might be useful tools to investigate the participation of cAMP in frog atrial contraction (unpublished observations).
Lugnier
C
C
Laboratoire de Pharmacologie Cellulaire et Moléculaire, Centre National de la Recherche Scientifique Unité 600, Université Louis Pasteur de Strasbourg, Illkirch, France.
Gauthier
C
C
Le Bec
A
A
Soustre
H
H
D015056:1-Methyl-3-isobutylxanthine / Q000494:pharmacology; D015105:3',5'-Cyclic-AMP Phosphodiesterases / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000378:metabolism*; D015106:3',5'-Cyclic-GMP Phosphodiesterases / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000378:metabolism*; D000818:Animals; D002122:Calcium Chloride / Q000494:pharmacology; D002147:Calmodulin / Q000494:pharmacology; D002852:Chromatography, Ion Exchange; D006152:Cyclic GMP / Q000494:pharmacology*; D003600:Cytosol / Q000201:enzymology; D004176:Dipyridamole / Q000494:pharmacology; D006325:Heart Atria; D006352:Heart Ventricles; D007700:Kinetics; D009206:Myocardium / Q000201:enzymology*; D010208:Papaverine / Q000494:pharmacology; D011688:Purinones / Q000494:pharmacology; D011760:Pyrrolidinones / Q000494:pharmacology; D011893:Rana esculenta; D011895:Rana ridibunda; D020889:Rolipram; D013379:Substrate Specificity; D013806:Theophylline / Q000494:pharmacology
D002147:Calmodulin; D011688:Purinones; D011760:Pyrrolidinones; D004176:Dipyridamole; D013806:Theophylline; D010208:Papaverine; D015105:3',5'-Cyclic-AMP Phosphodiesterases; D015106:3',5'-Cyclic-GMP Phosphodiesterases; C011145:zaprinast; D006152:Cyclic GMP; D020889:Rolipram; D002122:Calcium Chloride; D015056:1-Methyl-3-isobutylxanthine