Role of G protein beta gamma subunits in the regulation of the plasma membrane Ca2+ pump.
In Zajdela hepatoma cells (ZHC) the plasma membrane Ca2+ pump displayed no sensitivity to glucagon (19-29) (mini-glucagon), whereas in hepatocyte this metabolite of glucagon evoked a biphasic regulation of the Ca2+ pump system via a cholera toxin-sensitive G protein. Analysis of G protein subunits in ZHC membranes indicated the presence of cholera toxin-sensitive Gs alpha and G beta gamma proteins, whose functionality was manifested by GTP and NaF stimulation of adenylylcyclase activity, and pertussis toxin-catalyzed ADP-ribosylation of Gi alpha, respectively. However, immunoblotting experiments suggested a lower content in beta gamma subunits in ZHC as compared with hepatocyte plasma membranes. Complementation of ZHC or hepatocyte plasma membranes with purified beta gamma subunits from transducin (T beta gamma) caused inhibition of the basal activity of the Ca2+ pump at 10 and 300 ng/ml, respectively, and revealed (in ZHC) or increased (in hepatocytes) sensitivity of the system to mini-glucagon. After cholera toxin treatment of ZHC, T beta gamma no longer reconstituted the response of the Ca2+ pump to mini-glucagon, suggesting that the mechanism of beta gamma action is dependent on an association with the alpha subunit of a cholera toxin-sensitive G protein. It is concluded that G beta gamma subunits control both the basal activity of the plasma membrane Ca2+ pump and its inhibition by mini-glucagon.
Lotersztajn
S
S
Institut National de la Santé et la Recherche Médicale Unité 99, Hôpital Henri Mondor, Créteil, France.
Pavoine
C
C
Deterre
P
P
Capeau
J
J
Mallat
A
A
LeNguyen
D
D
Dufour
M
M
Rouot
B
B
Bataille
D
D
Pecker
F
F
D000246:Adenosine Diphosphate Ribose / Q000378:metabolism; D000262:Adenylyl Cyclases / Q000378:metabolism; D000818:Animals; D015153:Blotting, Western; D017301:Ca(2+) Mg(2+)-ATPase / Q000378:metabolism; D000252:Calcium-Transporting ATPases / Q000378:metabolism / Q000502:physiology*; D002461:Cell Line, Transformed; D002462:Cell Membrane / Q000201:enzymology / Q000378:metabolism; D002478:Cells, Cultured; D002772:Cholera Toxin / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D019204:GTP-Binding Proteins / Q000378:metabolism*; D005934:Glucagon / Q000378:metabolism; D006160:Guanosine Triphosphate / Q000494:pharmacology; D008099:Liver / Q000166:cytology / Q000201:enzymology / Q000378:metabolism; D051381:Rats; D012969:Sodium Fluoride / Q000494:pharmacology
D000246:Adenosine Diphosphate Ribose; D006160:Guanosine Triphosphate; D012969:Sodium Fluoride; D005934:Glucagon; D002772:Cholera Toxin; D017301:Ca(2+) Mg(2+)-ATPase; D019204:GTP-Binding Proteins; D000262:Adenylyl Cyclases; D000252:Calcium-Transporting ATPases