THE REDUCTION OF NEURONAL CALCIUM CURRENTS BY ATP-GAMMA-S IS MEDIATED BY A G-PROTEIN AND OCCURS INDEPENDENTLY OF CYCLIC AMP-DEPENDENT PROTEIN-KINASE

We studied the effects of ATP-gamma-S on the T, N and L calcium current components of nodose ganglion neurons using the whole cell variation of the patch clamp technique. ATP-gamma-S can serve as a phosphate donor in kinase-mediated reactions, the donated phosphate group being resistant to the action of phosphatases. We therefore compared the effect of ATP-gamma-S to that of the catalytic subunit of the cyclic AMP-dependent protein kinase (AK-C), included in the recording pipette with 5 mM ATP. AK-C (50 mu-g/ml) had no effect on the T current, and caused a approximately 30% increase in currents containing the N and L components during a 20-min recording, as compared to a approximately 45% decrease in control currents. In contrast, in the presence of 2.5 mM ATP-gamma-S, T currents declined approximately 30%, and currents containing the N and L components declined to a greater extent than control currents, about 65%. In addition, the time to peak current was increased from approximately 14% ms to approximately 40% ms. This effect of ATP-gamma-S on calcium currents was similar to that of certain neurotransmitters or GTP-gamma-S, an activator of G proteins, except that the effects of ATP-gamma-S were delayed 5-7 min relative to GTP-gamma-S. The effects of both ATP-gamma-S and GTP-gamma-S were reduced or abolished in neurons treated with pertussis toxin. We conclude that AK-C regulates neuronal calcium currents, presumably by phosphorylation of channels or associated proteins, and that the ATP-gamma-S-induced reduction of calcium currents cannot be due to its serving as a phosphate donor for endogenous AK. Instead, ATP-gamma-S reduces calcium currents via a G protein, perhaps by its conversion to GTP-gamma-S.