ADP-ribosylation of adenylate cyclase by pertussis toxin. Effects on inhibitory agonist binding.
Adenylate cyclase in NG108-15 (neuroblastoma X glioma hybrid) cells is responsive to both stimulatory and inhibitory ligands. Bordetella pertussis toxin (PT) catalyzes the ADP-ribosylation of a 41,000-Da peptide believed to be a subunit of the putative guanyl nucleotide-binding protein (Gi) involved in cyclase inhibition and abolishes inhibitory effects of opiate agonists. In studying the effects of PT on opiate receptors, we found that [3H]enkephalinamide binding was reduced by approximately 90% in membranes prepared from cells incubated with PT compared to control membranes. Agonist affinity, assessed by enkephalinamide competition for [3H]diprenorphine-binding sites, was markedly reduced in cells incubated with PT. Furthermore, inhibition by guanylylimidodiphosphate of ligand binding to opiate receptors was reduced following treatment with PT. The number of opiate receptors assessed by [3H]diprenorphine binding was unaltered by PT. These data are consistent with the hypothesis that PT-catalyzed ADP-ribosylation impairs the interaction of Gi with the inhibitory receptor-ligand complex, effectively uncoupling the inhibitory receptor from Gi and the cyclase catalytic unit.
D000246:Adenosine Diphosphate Ribose / Q000378:metabolism*; D037361:Adenylate Cyclase Toxin; D000262:Adenylyl Cyclases / Q000378:metabolism*; D000818:Animals; D001426:Bacterial Proteins / Q000494:pharmacology*; D004174:Diprenorphine / Q000378:metabolism; D004744:Enkephalin, Methionine / Q000031:analogs & derivatives / Q000378:metabolism; D006160:Guanosine Triphosphate / Q000494:pharmacology; D006165:Guanylyl Imidodiphosphate / Q000494:pharmacology; D051379:Mice; D009702:Nucleoside Diphosphate Sugars / Q000378:metabolism*; D037342:Pertussis Toxin; D011957:Receptors, Opioid / Q000378:metabolism; D012965:Sodium Chloride / Q000494:pharmacology; D010566:Virulence Factors, Bordetella
D037361:Adenylate Cyclase Toxin; D001426:Bacterial Proteins; D009702:Nucleoside Diphosphate Sugars; D011957:Receptors, Opioid; D010566:Virulence Factors, Bordetella; D004174:Diprenorphine; D000246:Adenosine Diphosphate Ribose; D006165:Guanylyl Imidodiphosphate; D012965:Sodium Chloride; D004744:Enkephalin, Methionine; C021055:Met-enkephalinamide; D006160:Guanosine Triphosphate; D037342:Pertussis Toxin; D000262:Adenylyl Cyclases