A cytosolic peptide potentiates the GTP effect on beta-adrenergic response of adenylate cyclase.
A cytosolic peptide-GTP complex that stimulates l-isoproterenol-responsive adenylate cyclase activity was identified in the rat liver. The peptide component was purified and characterized with regard to its interaction with GTP. The peptide was isolated as a complex form with GTP on a Sephadex G-25 column in 1 mM NaHCO3, and was purified as a dissociated form, with relative molecular weight (M(r)) approximately 3,000 and GTP-binding ability, in 200 mM ammonium acetate. The purified peptide alone displayed little stimulatory effect on adenylate cyclase activity, but its reassociated form with GTP clearly enhanced the effect of GTP on the enzyme activity. The isoproterenol competition curve using l-[3H]dihydroalprenolol as an antagonist ligand shifted to lower affinity by the addition of the peptide reassociated with GTP (16.5-fold shift), whereas the same concentration of GTP (1 microM) or the peptide alone had little or no effect (1.5- or 0.9-fold shift, respectively). Furthermore, the peptide enhanced the GTP effect in response to l-isoproterenol but not to glucagon, prostaglandin E1, or fluoride. These results suggest that the cytosolic peptide potentiates the effect of GTP on the agonist-beta-adrenergic receptor-stimulatory guanine nucleotide-binding regulatory component of the adenylate cyclase ternary complex.
Omori
K
K
Department of Pharmacology, Kansai Medical University, Osaka, Japan.
Kitagawa
K
K
Omori
K
K
Yasukura
T
T
Uriu
T
T
Morita
M
M
Inagaki
C
C
D000262:Adenylyl Cyclases / Q000378:metabolism*; D000318:Adrenergic beta-Agonists / Q000494:pharmacology*; D000818:Animals; D001667:Binding, Competitive; D003600:Cytosol / Q000378:metabolism*; D004082:Dihydroalprenolol / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation; D005260:Female; D006160:Guanosine Triphosphate / Q000378:metabolism / Q000502:physiology*; D007545:Isoproterenol / Q000494:pharmacology; D010455:Peptides / Q000502:physiology*; D051381:Rats; D017208:Rats, Wistar
D000318:Adrenergic beta-Agonists; D010455:Peptides; D004082:Dihydroalprenolol; D006160:Guanosine Triphosphate; D000262:Adenylyl Cyclases; D007545:Isoproterenol