Reduction in the stability of the Gs-catalytic unit complex of adenylate cyclase in isoproterenol-induced heterologous desensitization.
Treatment of rat reticulocytes with isoproterenol resulted in the heterologous desensitization of adenylate cyclase, causing a reduction in NaF/AlCl3- and guanylyl 5'-imidodiphosphate (Gpp(NH)p)-stimulated activities as well as in activity stimulated by beta-adrenoceptor agonists. Desensitization was also induced by dibutyryl cyclic AMP and 1-(5-isoquinolinylsulfonyl)-2-methylpiperazine (H-7), an inhibitor of cyclic AMP-dependent protein kinase (protein kinase A), prevented the isoproterenol-induced desensitization, suggesting the involvement of protein kinase A in the desensitization. Gs in the desensitized cell membrane was activated by treatment with Gpp(NH)p or NaF/AlCl3 in the presence of Mg2+. The activated state, the Gs-C complex, was gradually shifted to the basal state, i.e. the Gs-C complex was dissociated by removal of Mg2+. The rate constant of the dissociation was increased in the desensitized cell membranes (0.074 min-1) as compared with that in the control cell membranes (0.022 min-1). The half life of the Gs-C complex, calculated from the rate constant, was decreased during the process of desensitization. The results indicate that reduction in the stability of the Gs-C complex is related to heterologous desensitization. Agents which increase the level of cyclic AMP in the cell also reduced the stability of the complex. The potency of such an effect was in the following order; isoproterenol approximately NaF/AlCl3 (k = 0.071 min-1) greater than forskolin (0.039 min-1) greater than Mn2+ approximately dibutyryl cyclic AMP (0.030 min-1). The former two activate Gs and form Gs-C complex, while the latter two cause intracellular cyclic AMP accumulation without activation of Gs. It may be deduced from this order that the reduction of the stability depends not only on the increase in the cyclic AMP level but also on the level of Gs-C complex formed.
Yamashita
A
A
Department of Physiological Chemistry, Hiroshima University School of Medicine, Japan.
Kurokawa
T
T
Une
Y
Y
Ishibashi
S
S
D000262:Adenylyl Cyclases / Q000378:metabolism*; D000818:Animals; D001798:Blood Proteins / Q000032:analysis; D002384:Catalysis; D002462:Cell Membrane / Q000201:enzymology; D000242:Cyclic AMP / Q000032:analysis; D004795:Enzyme Stability / Q000187:drug effects; D066298:In Vitro Techniques; D007545:Isoproterenol / Q000494:pharmacology*; D007700:Kinetics; D008297:Male; D011863:Radioimmunoassay; D051381:Rats; D011919:Rats, Inbred Strains; D012156:Reticulocytes / Q000187:drug effects / Q000201:enzymology
D001798:Blood Proteins; D000242:Cyclic AMP; D000262:Adenylyl Cyclases; D007545:Isoproterenol