Ca(2+) elevation evoked by membrane depolarization regulates G protein cycle via RGS proteins in the heart.
Regulators of G protein signaling (RGS), which act as GTPase activators, are a family of cytosolic proteins emerging rapidly as an important means of controlling G protein-mediated cell signals. The importance of RGS action has been verified in vitro for various kinds of cell function. Their in situ modes of action in intact cells are, however, poorly understood. Here we show that an increase in intracellular Ca(2+) evoked by membrane depolarization controls the RGS action on G protein activation of muscarinic K(+) (K(G)) channel in the heart. Acetylcholine-induced K(G) current exhibits a slow time-dependent increase during hyperpolarizing voltage steps, referred to as "relaxation." This reflects the relief from the decrease in available K(G) channel number induced by cell depolarization. This phenomenon is abolished when an increase in intracellular Ca(2+) is prevented. It is also abolished when a calmodulin inhibitor or a mutant RGS4 is applied that can bind to calmodulin but that does not accelerate GTPase activity. Therefore, an increase in intracellular Ca(2+) and the resultant formation of Ca(2+)/calmodulin facilitate GTPase activity of RGS and thus decrease the available channel number on depolarization. These results indicate a novel and probably general pathway that Ca(2+)-dependent signaling regulates the G protein cycle via RGS proteins.
Ishii
M
M
Department of Pharmacology II , Graduate School of Medicine, Osaka University, Suita, Osaka, Japan.
Inanobe
A
A
Fujita
S
S
Makino
Y
Y
Hosoya
Y
Y
Kurachi
Y
Y
D000200:Action Potentials; D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002147:Calmodulin / Q000037:antagonists & inhibitors; D002478:Cells, Cultured; D002614:Chelating Agents / Q000494:pharmacology; D004533:Egtazic Acid / Q000031:analogs & derivatives* / Q000494:pharmacology; D004553:Electric Conductivity; D006321:Heart / Q000502:physiology*; D020962:Heterotrimeric GTP-Binding Proteins / Q000378:metabolism*; D017136:Ion Transport; D007700:Kinetics; D009154:Mutation; D009206:Myocardium / Q000166:cytology / Q000378:metabolism; D024642:Potassium Channels, Voltage-Gated / Q000037:antagonists & inhibitors / Q000502:physiology*; D020710:RGS Proteins / Q000235:genetics / Q000502:physiology*; D051381:Rats; D011921:Rats, Inbred WKY
D002147:Calmodulin; D002614:Chelating Agents; D024642:Potassium Channels, Voltage-Gated; D020710:RGS Proteins; C098799:RGS4 protein; D004533:Egtazic Acid; D020962:Heterotrimeric GTP-Binding Proteins; C025603:1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid; D002118:Calcium