Association of a 68,000-dalton protein with adrenocorticotropin-sensitive adenylate cyclase activity in Y1 adrenocortical tumor cells.
This report explores the biochemical basis for clonal variation in adrenocorticotropin (ACTH)-sensitive adenylate cyclase activity in the Y1 mouse adrenocortical tumor cell line. We demonstrate that the level of a specific protein, designated p68, is significantly correlated with the ability of adrenocorticotropin to stimulate adenylate cyclase activity among Y1 subclones (p = 0.004; r = 0.65). p68 was characterized by its molecular weight in sodium dodecyl sulfate polyacrylamide gels (Mr = 68,000) and by its isoelectric point as determined by two-dimensional gel electrophoresis (pI = 7.2). On two-dimensional gels, the protein migrated as a major spot with satellite spots 0.1 pH unit on either side. Homogenates and plasma membrane fractions from clones highly responsive to ACTH had large amounts of p68. In homogenates from highly responsive clones p68 represented 10 to 12% of the total protein. Homogenates and plasma membrane fractions from clones insensitive to ACTH were deficient in p68. In homogenates from the insensitive clones Y6 and OS3, p68 represented less than or equal 0.8% of the total protein. A somatic cell hybrid, formed by fusion of these two ACTH-insensitive clones recovered ACTH-sensitive adenylate cyclase activity and concomitantly expressed appreciable levels of p68. It is suggested that p68 may regulate the transfer of information from the occupied ACTH receptor ot the catalytic subunit of adenylate cyclase.
D000262:Adenylyl Cyclases / Q000378:metabolism*; D000306:Adrenal Cortex Neoplasms / Q000201:enzymology*; D000324:Adrenocorticotropic Hormone / Q000494:pharmacology*; D000818:Animals; D002460:Cell Line; D002462:Cell Membrane / Q000201:enzymology; D002999:Clone Cells; D051379:Mice; D008970:Molecular Weight; D009363:Neoplasm Proteins / Q000502:physiology*; D009374:Neoplasms, Experimental / Q000201:enzymology
D009363:Neoplasm Proteins; D000324:Adrenocorticotropic Hormone; D000262:Adenylyl Cyclases