Receptor-G-protein coupling in desensitization-resistant mutant adrenal cells.
Mutant isolates from the Y1 mouse adrenocortical tumor cell line (designated Y1DR) were previously shown to resist homologous desensitization of adenylyl cyclase via endogenous ACTH receptors or transfected beta 2-adrenergic receptors. To further localize the site of the DR mutation, we examined the agonist-binding properties of the transfected beta 2-adrenergic receptor in parental Y1 cells and the DR mutant before and after desensitization with the beta-adrenergic agonist isoproterenol. Binding studies were carried out using [125I]iodocyanopindolol as the labeled ligand and isoproterenol as the competing agonist. We found that the DR mutant has a greater number of high affinity receptors in the basal state than does the Y1 parent (70-80% vs. < or = 50%) and that treatment of membranes from parent or mutant cells with guanyl-5'-yl imidodiphosphate and NaCl converts all of the high affinity sites to low affinity sites. After desensitization with isoproterenol, only low affinity binding sites are detected in parental Y1 cells, whereas the DR mutants retain an appreciable number of high affinity receptors. These results indicate that the DR mutant may resist desensitization by affecting agonist-induced uncoupling of receptors from their guanyl nucleotide-binding regulatory G-proteins.
Olson
M F
MF
Banting and Best Department of Medical Research, University of Toronto, Ontario, Canada.
Schimmer
B P
BP
D000311:Adrenal Glands / Q000378:metabolism* / Q000473:pathology; D000318:Adrenergic beta-Agonists / Q000378:metabolism; D000818:Animals; D001665:Binding Sites; D019204:GTP-Binding Proteins / Q000378:metabolism*; D006165:Guanylyl Imidodiphosphate / Q000494:pharmacology; D006706:Homeostasis / Q000502:physiology*; D020120:Iodocyanopindolol; D007545:Isoproterenol / Q000494:pharmacology; D051379:Mice; D009154:Mutation; D010869:Pindolol / Q000031:analogs & derivatives / Q000378:metabolism; D011943:Receptors, Adrenergic, beta / Q000378:metabolism*; D014407:Tumor Cells, Cultured
D000318:Adrenergic beta-Agonists; D011943:Receptors, Adrenergic, beta; D006165:Guanylyl Imidodiphosphate; D020120:Iodocyanopindolol; D010869:Pindolol; D019204:GTP-Binding Proteins; D007545:Isoproterenol