Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: EC:4.6.1.1 (adenylate cyclase)
 19,190 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

The radioiodinated beta-blocker iodohydroxybenzylpindolol ([125I]HYP) has been used to identify directly and characterize beta-adrenergic receptors in isolated bovine parathyroid cells. [125I]HYP was bound rapidly and reversibly to isolated bovine parathyroid cell membranes. Scatchard analysis revealed a single class of binding sites with high affinity (4 X 10(10M-1) and low capacity (0.7 pmol/mg). Saturation analysis of [125I]HYP binding to intact bovine parathyroid cells suggested a site with similar affinity on whole cells and with a binding capacity of 5000-10,000 sites/cell. True dissociation constants (Kd's) for beta-adrenergic agonists and antagonists were in good agreement with activation constants (KA'S) and inhibition constants (KI'S) for effects on adenylate cyclase in membrane preparations. These constants also were in reasonable agreement with KA'S and KI'S previously shown for effects of agonists and antagonists on cAMP accumulation and PTH release in whole cells. This study shows by direct analysis that beta-adrenergic receptors exist on isolated bovine parathyroid cells, and that there is close coupling between receptor binding, effects on cAMP and hormonal release. This represents still another system in which [125I]HYP has been successfully used to study beta adrenergic receptors in membrane as well as intact cell preparations.
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PMID:Direct identification of beta-adrenergic receptors on isolated bovine parathyroid cells. 1 24

Turkey erythrocyte membranes contain beta-adrenergic-coupled adenylate cyclase systems that are modulated by guanine nucleotides. Incubation of membranes with Gpp (NH) p plus isoproterenol led to a persistently activated state ("holocatalytic state") of adenylate cyclase independent of agonist. Formation of this holocatalytic state was inhibited by conditions (4 degrees) or by compounds (e.g., GTP EDTA) that prevented Gpp (NH) p binding or that prevented binding of isoproterenol (e.g., propranolol). None of these agents, however, reduced activity of this activated state once it had been formed. The holocatalytic state, even though resistant to inhibition by propranolol, showed no change in receptor affinity or number of sites as determined by binding of 125I-HYP, a high affinity beta-adrenergic antagonist. Formation of the holocatalytic state, therefore, involves a modification of the adenylate cyclase system distal to the hormone receptor complex.
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PMID:Holocatalytic state of adenylate cyclase in turkey erythrocyte membranes: formation with guanylylimidodiphosphate plus isoproterenol without effect on affinity of beta-receptor. 81 36

Iodohydroxybenzylpindolol (I-HYP) is a chemically defined, high affinity, high specific activity beta-adrenergic antagonist that interacts with a single site on the turkey erythrocyte membrane. Study of the interaction of agonists, antagonists, and congeners with this site and concomitant alterations in adenylate cyclase activity have been carried out in the presence of high or low concentrations of guanine nucleotide. The results help clarify the relationship between binding and activation or inhibition of adenylate cyclase and the role of guanine nucleotides in modulating this interaction. There is a close correlation between binding constants (KD) for inhibitors determined by analysis of competitive displacement of 125I-HYP from receptor, and apparent affinities (Ki) for inhibition of adenylate cyclase. For activators, however, there is up to a 10-fold difference between KD and apparent affinity (KDapp) for adenylate cyclase activation at low guanine nucleotide concentration (10(-6) M guanylylimidodiphosphate). This difference is virtually abolished by employing higher nucleotide concentrations (10(-5) M guanylylimidodiphosphate) without significantly altering receptor affinity. This suggests that guanine nucleotides act by modulating receptor-enzyme interactions rather than hormone-receptor interactions. Moreover, several beta-adrenergic analogs previously shown to have no effect on adenylate cyclase in the absence of nucleotide, are partial agonists in the presence of 10(-5) M guanylylimidodiphosphate. Parallel analyses for a series of agonists and antagonists for adenylate cyclase activation and receptor interaction show affinities for levorotatory isomers generally 100-fold greater than for dextrorotatory isomers. Thus stereoconfiguration at the beta carbon clearly influences affinity of agonists or antagonists. Affinity is also importantly influenced by the nature of the aromatic ring as well as the N-alkyl group. The complexity of structure-function relationships for these compounds requires a redefinition of structural requirements for beta-adrenergic activity.
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PMID:Beta-Adrenergic receptor interactions. Direct comparison of receptor interaction and biological activity. 125 66

beta-Adrenergic receptors and catecholamine-sensitive adenylate cyclase were identified and partially characterized in membrane fractions of rabbit lungs from day 25 of gestation to adulthood with the beta-adrenergic antagonists (--)-[3H]dihydroalprenolol [(--)-[3H]DHA] and (--)-[125I]iodohydroxybenzylpindolol [(--)-[125I]HYP]. beta-Adrenergic receptor number (Bmax) increased 11.5-fold during this time period, increasing progressively during the latter days of gestation and the early neonatal period, from 37 +/- 10 fmol/mg protein at 25 days gestation to 425 +/- 51 fmol/mg in the adult rabbit lung (mean +/- SD). Receptor affinity for (--)-[3H]DHA (KD = 1.8 nM) or (--)-[125I]HYP (KD - 0.104 nM) and the proportion of beta 1- and beta 2-adrenergic receptor subtypes (60% beta 1 and 40% beta 2) did not change with advancing age. Basal adenylate cyclase activity in lung homogenates decreased significantly with increasing age, whereas the activity in the presence of catecholamine or NaF remained nearly constant. Catecholamines stimulated adenylate cyclase activity at all ages studied supporting a role of the maturation of beta-adrenergic receptors in the regulation of pulmonary function.
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PMID:beta-Adrenergic receptors in the developing rabbit lung. 626 84

Interaction of the insecticide 1,1,1-trichloro-2,2-bis-(p-chlorophenyl)-ethane (DDT) with beta-receptor binding and adenylate cyclase activity of biological membranes has been studied. Following exposure of cultured Chang liver cells to DDT, maximal binding of the catecholamine antagonist [125I]-iodohydroxybenzylpindolol (HYP) to isolated cell membranes was decreased by 30% whereas the dissociation constant remained unchanged. Both basal activity and maximal isoproterenol-stimulated activity of adenylate cyclase were not altered. The isoproterenol concentration required for half-maximal stimulation of the enzyme was increased about 2-fold as was the agonist concentration required for half-maximal displacement of the antagonist HYP at the beta-receptor binding site. Thus, coupling efficiency of hormone-stimulated adenylate cyclase activity was not influenced by the presence of DDT in these membranes. The data show that interaction of DDT with the beta-receptor adenylate cyclase complex is restricted to the receptor component. Enzyme activity is directly linked to changes of agonist binding at the beta-receptor. Interference of DDT with signal transduction via 'fluidization' of membrane lipids has not been detected.
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PMID:Interaction of the insecticide 1,1,1-trichloro-2,2-bis(p-chlorophenyl)ethane with the beta-receptor adenylate cyclase system in Chang liver cell membranes. 631 12

We compared the properties of the adenylate cyclase system in plasma membranes derived from gas cavitation and sonication of human neutrophils. In membranes prepared from cavitated cells, cyclase was stimulated by fluoride ion and Gpp(NH)p. Stimulation by isoproterenol (and PGE1) was absent, although the beta-receptor was present as judged by 125I-HYP binding. Two unusual characteristics of this cyclase system are a) substantial activation of cyclase by GTP in the absence of hormone, and b) reduction in the regulation of the beta-receptor by GTP. By contrast, vesicles isolated from sonicated cells displayed normal GTP-dependent activation by isoproterenol (as well as PGE1) and GTP regulation of the beta-receptor, while hormone-independent stimulation by GTP was negligible. Cholera toxin-catalyzed ADP ribosylation revealed a prominent band at 42K in both membranes, and a minor band at 35K, although the degree of labeling of this protein was lower in the "cavitated" vesicles. Our results suggest that the mode of cell lysis and membrane preparation influence receptor-cyclase interactions and that receptor-cyclase uncoupling is associated with amplified GTP activation and altered receptor regulation by GTP.
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PMID:The beta-adrenergic receptor in the human neutrophil plasma membrane: receptor-cyclase uncoupling is associated with amplified GTP activation. 631 94

We have measured the beta-adrenergic receptor content in 1- to 10-day-old corpora lutea. Corpora lutea of defined ages were obtained by sc injection of 8 IU PMSG to 26-day-old rats, leading to ovulation and formation of corpora lutea in the early morning of day 29. Membrane fractions of isolated corpora lutea were incubated with various concentrations of the beta-adrenergic antagonist [125I]iodohydroxybenzylpindolol [125I]iodo-HYP, 12.5-2500 pM) for 60 min at 22 C. Alprenolol (10(-5) M) was used to determine nonspecific binding. Bound and free [125I]iodo-HYP was separated by filtration and washing on Whatman GF/C filters under vacuum. There was a 3-fold increase in beta-adrenergic receptor concentration during the first 2-3 days of corpus luteum formation, followed by a decline in the beta-adrenergic receptor content with luteal age. The rat luteal beta-adrenergic receptor seems to be of the beta 2-subtype as determined from adenylate cyclase stimulation as well as from displacement of [125I]iodo-HYP binding by various beta-adrenergic agonists.
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PMID:Beta-adrenergic receptor concentration in corpora lutea of different ages obtained from pregnant mare serum gonadotropin-treated rats. 632 35