Receptor and effector interactions of Gs. Functional studies with antibodies to the alpha s carboxyl-terminal decapeptide.
Antibodies generated to a synthetic decapeptide, RMHLRQYELL, representing the carboxyl-terminus of Gs-alpha have been characterized in immunoblots and functional studies. This antibody, designated RM, reacts exclusively with a doublet of proteins of 52 and 45 kDa in immunoblots of bovine brain and wild-type S49 murine lymphoma cell membranes. No such reactivity is seen in membranes from cyc- S49 cells, which lack Gs. RM blocks receptor-mediated activation of Gs and adenylyl cyclase in membranes from wild-type S49 cells. RM could also immunoprecipitate adenylyl cyclase activity in detergent extracts from GTP[gamma]S- or fluoride-preactivated bovine brain membranes; thus binding of alpha s to effector and carboxyl-terminal antibody was mutually compatible. Such experiments provide an approach for the elucidation of functionally relevant interactions of G-proteins with receptors and effectors in the membrane.
Simonds
W F
WF
Molecular Pathophysiology Branch, National Institute of Diabetes, Digestive and Kidney Diseases, Bethesda, MD 20892.
Goldsmith
P K
PK
Woodard
C J
CJ
Unson
C G
CG
Spiegel
A M
AM
D000262:Adenylyl Cyclases / Q000378:metabolism; D000818:Animals; D000906:Antibodies / Q000276:immunology; D015153:Blotting, Western; D001921:Brain / Q000201:enzymology; D002417:Cattle; D004591:Electrophoresis, Polyacrylamide Gel; D019204:GTP-Binding Proteins / Q000276:immunology / Q000378:metabolism*; D051379:Mice; D010455:Peptides / Q000276:immunology*; D011817:Rabbits; D014407:Tumor Cells, Cultured
D000906:Antibodies; D010455:Peptides; D019204:GTP-Binding Proteins; D000262:Adenylyl Cyclases