Voltage-dependent potentiation of the activity of cardiac L-type calcium channel alpha 1 subunits due to phosphorylation by cAMP-dependent protein kinase.
Barium currents mediated by the alpha 1 subunit of the cardiac L-type Ca channel expressed in Chinese hamster ovary (CHO) cells were increased up to 10-fold during dialysis of the cell with the catalytic subunit of cAMP-dependent protein kinase. After partial activation by exogenous kinase, the activity of the alpha 1 subunit was also reversibly potentiated up to 3.5-fold by prepulses to voltages in the range of 0 to +150 mV. Potentiation at +48 mV developed with a biphasic time course with time constants of 131 ms and 8 s. Reversal at -60 mV was biphasic with half-times of 12 ms and 100 ms and was blocked in the presence of the phosphatase inhibitor okadaic acid. Both the increase in calcium-channel activity during dialysis with kinase and the voltage-dependent potentiation were accompanied by shifts in the voltage dependence of activation to more negative membrane potentials. The increases in Ba current due to protein phosphorylation and to the dihydropyridine Ca channel agonist Bay K8644 were approximately additive. The results show that the alpha 1 subunit of the cardiac L-type Ca channel is sufficient for substantial modulation of Ca-channel activity by cAMP-dependent protein kinase and for potentiation by state-dependent protein phosphorylation. Voltage-dependent potentiation of the activity of the alpha 1 subunit may contribute to the increase in contractile force in response to increased rate of stimulation, the positive staircase effect in heart muscle.
Sculptoreanu
A
A
Department of Pharmacology, University of Washington, Seattle 98195.
Rotman
E
E
Takahashi
M
M
Scheuer
T
T
Catterall
W A
WA
D001498:3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethyl-5-nitro-4-(2-(trifluoromethyl)phenyl)-, Methyl ester / Q000494:pharmacology*; D000818:Animals; D001464:Barium / Q000378:metabolism; D016466:CHO Cells; D015220:Calcium Channels / Q000187:drug effects / Q000378:metabolism / Q000502:physiology*; D006224:Cricetinae; D017868:Cyclic AMP-Dependent Protein Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D004988:Ethers, Cyclic / Q000494:pharmacology; D005071:Evoked Potentials / Q000187:drug effects; D006321:Heart / Q000502:physiology; D046911:Macromolecular Substances; D008564:Membrane Potentials; D009206:Myocardium / Q000378:metabolism*; D019319:Okadaic Acid; D010766:Phosphorylation; D011994:Recombinant Proteins / Q000187:drug effects / Q000378:metabolism; D013997:Time Factors; D014162:Transfection
D015220:Calcium Channels; D004988:Ethers, Cyclic; D046911:Macromolecular Substances; D011994:Recombinant Proteins; D019319:Okadaic Acid; D001464:Barium; D001498:3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethyl-5-nitro-4-(2-(trifluoromethyl)phenyl)-, Methyl ester; D017868:Cyclic AMP-Dependent Protein Kinases