Intracellular ATP and GTP are both required to preserve modulation of N-type calcium channel current by norepinephrine.
Norepinephrine (NE) inhibits voltage-dependent calcium channels of sympathetic neurons. We investigated the role of intracellular nucleotides in this inhibition for clues to receptor-channel coupling mechanisms. Both ATP and GTP are required to preserve NE responsiveness during whole-cell dialysis. The response to NE was gradually lost in bullfrog sympathetic neurons dialyzed with GTP as the only nucleotide, ATP only, or no nucleotides. Replacing ATP with ATP[gamma-S] resulted in spontaneous modulation of calcium channel current, possibly because of production of GTP[gamma-S]. The nonhydrolyzable ATP analog p[NH]ppA could substitute for ATP to preserve NE responsiveness. The protein phosphatase inhibitors okadaic acid and calyculin-A did not affect NE inhibition of calcium channel current, or recovery from that inhibition. These results suggest protein phosphorylation is not involved in the inhibition of calcium channel current, but binding of ATP to some intracellular site is required for the coupling of adrenergic receptors to calcium channels.
Department of Physiology and Biophysics, Case Western Reserve University, Cleveland, Ohio 44106.
K S
KS
Elmslie
M A
MA
Werz
J L
JL
Overholt
S W
SW
Jones
D000255:Adenosine Triphosphate / Q000031:analogs & derivatives / Q000494:pharmacology / Q000502:physiology*; D000818:Animals; D001665:Binding Sites / Q000187:drug effects; D015220:Calcium Channels / Q000187:drug effects / Q000378:metabolism*; D004988:Ethers, Cyclic / Q000494:pharmacology; D016244:Guanosine 5'-O-(3-Thiotriphosphate) / Q000494:pharmacology; D006160:Guanosine Triphosphate / Q000031:analogs & derivatives / Q000502:physiology*; D066298:In Vitro Techniques; D007700:Kinetics; D008387:Marine Toxins; D009474:Neurons / Q000187:drug effects / Q000378:metabolism; D009638:Norepinephrine / Q000494:pharmacology*; D019319:Okadaic Acid; D010080:Oxazoles / Q000494:pharmacology; D010749:Phosphoprotein Phosphatases / Q000037:antagonists & inhibitors; D011892:Rana catesbeiana; D013564:Sympathetic Nervous System / Q000166:cytology / Q000378:metabolism; D014662:Vasoconstrictor Agents / Q000494:pharmacology
D015220:Calcium Channels; D004988:Ethers, Cyclic; D008387:Marine Toxins; D010080:Oxazoles; D014662:Vasoconstrictor Agents; D019319:Okadaic Acid; C022571:adenosine 5'-O-(3-thiotriphosphate); D016244:Guanosine 5'-O-(3-Thiotriphosphate); C059041:calyculin A; D006160:Guanosine Triphosphate; D000255:Adenosine Triphosphate; D010749:Phosphoprotein Phosphatases; D009638:Norepinephrine