cAMP and calcium-dependent mechanisms of phospholamban phosphorylation in intact hearts.
The present work was undertaken with two main goals: 1) to further elucidate the physiological role of the adenosine 3',5'-cyclic monophosphate (cAMP) and Ca2(+)-calmodulin (Ca2(+)-Cm)-dependent mechanisms of phospholamban phosphorylation (32PiPHL), and 2) to study the possible interaction between these two systems in the intact heart. Interventions that increased twitch or tetanic tension without modifying cAMP levels [high extracellular Ca2+ concentration [( Ca2+]o) or BAY K 8644 in catecholamine-depleted hearts] failed to alter 32PiPHL. Moderate and high beta-adrenergic stimulation (3 x 10(-9) and 3 x 10(-8) M isoproterenol, respectively) increased cAMP from 0.345 +/- 0.032 to 0.636 +/- 0.069 and 0.772 +/- 0.060 pmol/mg wet wt, and 32PiPHL from 26.8 +/- 4.1 to 58.6 +/- 13.1 and 174.7 +/- 13.8 pmol 32Pi/mg sarcoplasmic reticular [SR] protein, respectively. Both doses of isoproterenol produced an enhanced myocardial relaxation. Reversal of the positive inotropic effect of isoproterenol by interventions that decrease intracellular Ca2+ supply failed to reduce the enhancement in 32PiPHL and myocardial relaxation elicited by 3 x 10(-9) M isoproterenol but diminished the increase in 32PiPHL induced by 3 x 10(-8) M isoproterenol to 116.3 +/- 10.9 without significant changes in cAMP. Changes in myocardial relaxation closely paralleled the changes in 32PiPHL. These results suggest that 1) 32PiPHL may be enhanced by the cAMP-dependent mechanism independently of the Ca2(+)-Cm system, and 2) 32PiPHL and myocardial relaxation may be modified by intracellular Ca2+ changes only at high-intracellular cAMP levels.
Centro de Investigaciones Cardiovasculares, Facultad de Ciencias Medicas, La Plata, Argentina.
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L
Vittone
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Mundiña
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Chiappe de Cingolani
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A
Mattiazzi
D001498:3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethyl-5-nitro-4-(2-(trifluoromethyl)phenyl)-, Methyl ester / Q000494:pharmacology; D000251:Adenosine Triphosphatases / Q000378:metabolism; D000818:Animals; D001696:Biomechanical Phenomena; D002118:Calcium / Q000502:physiology*; D002135:Calcium-Binding Proteins / Q000378:metabolism*; D002147:Calmodulin / Q000502:physiology; D000242:Cyclic AMP / Q000378:metabolism / Q000502:physiology*; D066298:In Vitro Techniques; D007425:Intracellular Membranes / Q000378:metabolism; D007545:Isoproterenol / Q000494:pharmacology; D008297:Male; D009200:Myocardial Contraction / Q000187:drug effects; D009206:Myocardium / Q000166:cytology / Q000378:metabolism*; D009543:Nifedipine / Q000494:pharmacology; D009994:Osmolar Concentration; D010766:Phosphorylation; D051381:Rats; D011919:Rats, Inbred Strains; D000099211:Phospholamban
D001498:3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethyl-5-nitro-4-(2-(trifluoromethyl)phenyl)-, Methyl ester; D000251:Adenosine Triphosphatases; D002118:Calcium; D002135:Calcium-Binding Proteins; D002147:Calmodulin; D000242:Cyclic AMP; D007545:Isoproterenol; D009543:Nifedipine; D000099211:Phospholamban