Intracellular protein kinase and calcium inward currents in perfused neurones of the snail Helix pomatia.
Changes in the amplitude of the calcium inward current caused by intracellular administration of tolbutamide (an inhibitor of the cyclic AMP-dependent protein kinase activity) or catalytic subunits of cAMP-dependent protein kinases from rabbit myocardium were studied on internally perfused nerve cells of the snail, Helix pomatia. Intracellular administration of 7 mM tolbutamide caused a rapid decline of the amplitude of the calcium current that had been stabilized by theophylline; the effect was practically completely reversible. In contrast, addition to the perfusing solution of exogenous catalytic subunits of cyclic AMP-dependent protein kinases (about 0.7 microM of protein) together with 2 mM adenosine 5'-triphosphate and 3 mM MgCl2, led to stabilization of the calcium conductance of the cell membrane or restored it if it had declined during the perfusion with basic solution. The effect depended largely on the presence of adenosine 5'-triphosphate. Its time course was very slow (dozens of minutes) due probably to slow diffusion of the protein inside the cell. Heat-inactivated catalytic subunits did not produce such a stabilizing or restoring action on the calcium conductance. The results substantiate the suggestion that the normal functioning of calcium channels depends on phosphorylation catalyzed by cyclic AMP-dependent protein kinases.
D000255:Adenosine Triphosphate / Q000502:physiology; D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002384:Catalysis; D006372:Helix, Snails / Q000502:physiology; D066298:In Vitro Techniques; D007473:Ion Channels / Q000502:physiology*; D009474:Neurons / Q000502:physiology*; D011188:Potassium / Q000378:metabolism; D047428:Protein Kinase Inhibitors; D011494:Protein Kinases / Q000502:physiology*; D014044:Tolbutamide / Q000494:pharmacology
D007473:Ion Channels; D047428:Protein Kinase Inhibitors; D000255:Adenosine Triphosphate; D014044:Tolbutamide; D011494:Protein Kinases; D011188:Potassium; D002118:Calcium