Activated Gs alpha but not Gi alpha prevents the thermal inactivation of adenylyl cyclase in plasma membranes derived from S49 lymphoma cells.
The thermal inactivation of adenylyl cyclase was studied in plasma membranes isolated from wild-type and the mutant cell strain cyc- of S49 lymphoma. The half-life of adenylyl cyclase activity at 30 degrees C was decreased from 14.2 min to 3.4 min by the presence of detergents. ATP as well as forskolin prevented the adenylyl cyclase inactivation in a dose-response manner independent of the utilized type of cell membranes. Activation of G-proteins by GTP gamma S or by AlF-4 in wild-type membranes but not in cyc- membranes partially prevented adenylyl cyclase inactivation. Adenylyl cyclase activity in cyc- membranes was preserved in the presence of GTP gamma S or AlF-4 from the observed detergent-induced inactivation by complementation of these membranes with an extract from wild-type membranes. ADP-ribosylation of Gi alpha in cyc- membranes did not influence the kinetics of the inactivation process of adenylyl cyclase, whereas ADP-ribosylated Gs alpha protein protected adenylyl cyclase more effectively than non-ribosylated Gs alpha in wild-type plasma membranes when GTP was used as an activator.
Kvapil
P
P
Wallenberg Laboratory for Cardiovascular Research, University of Gothenburg, Sweden.
Novotny
J
J
Ransnäs
L A
LA
D000255:Adenosine Triphosphate / Q000378:metabolism; D000067956:Adenylyl Cyclase Inhibitors; D000262:Adenylyl Cyclases / Q000378:metabolism*; D002462:Cell Membrane / Q000201:enzymology*; D005576:Colforsin / Q000494:pharmacology; D003902:Detergents / Q000494:pharmacology; D004789:Enzyme Activation; D019204:GTP-Binding Proteins / Q000378:metabolism*; D008223:Lymphoma; D013696:Temperature; D014407:Tumor Cells, Cultured
D000067956:Adenylyl Cyclase Inhibitors; D003902:Detergents; D005576:Colforsin; D000255:Adenosine Triphosphate; D019204:GTP-Binding Proteins; D000262:Adenylyl Cyclases