Problems in identification of the beta-adrenergic receptor.

Haber, Edgar; Wrenn, Simeon M.

doi:10.1152/physrev.1976.56.2.317

Cited by 39 publications

(7 citation statements)

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“…1). This gel exhibits major advantages over compounds described previously (17,18). In contrast with norepinephrine linked to agarose through an amido bond, no catechol group projects from the gel, thus avoiding retention of catechol acceptors; the amino group of the ethanolamine side chain, which is important for receptor recognition, remains free.…”

Section: Discussionmentioning

confidence: 92%

Isolation of adenylate cyclase-free, beta-adrenergic receptor from turkey erythrocyte membranes by affinity chromatography.

Vauquelin¹,

Geynet²,

Hanoune³

et al. 1977

Proc. Natl. Acad. Sci. U.S.A.

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The adenylate cyclase [ATP pyrophosphatelyase (cyclizing) EC 4.6.1.11 and ,-adrenergic receptor of plasma membranes of turkey erythrocytes were solubilized in an active form by treatment with either NaF or guanylylimidodiphosphate and digitonin. The solubilized enzyme was no longer stimulated by catecholamines, NaF, or guanine nucleotides. The digitonin extract was chromatographed on an alprenolol-agarose derivative. While the bulk of protein and all the adenylate cyclase activity passed unretarded through the column, the receptor was retained. It eluted free of enzyme activity with an alprenolol solution containing 1 M NaCl; the yield was 25-30%. The protein content of the alprenolol eluates was too low to be estimated by the Lowry technique and was assessed by a more sensitive fluorometric method. Under these conditions, the ,Badrenergic receptor was purified approximately 2000-fold in a single step with retention of all its pharmacological properties.These experiments establish that the f-adrenergic receptor and the adenylate cyclase are independent entities which may be separated on a functional basis.

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Section: Discussionmentioning

confidence: 92%

Isolation of adenylate cyclase-free, beta-adrenergic receptor from turkey erythrocyte membranes by affinity chromatography.

Vauquelin¹,

Geynet²,

Hanoune³

et al. 1977

Proc. Natl. Acad. Sci. U.S.A.

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“…f-adrenergic actions in various tissues including erythrocytes have been considered to be mediated by increased intracellular cAMP concentrations by activation of adenylate cyclase (31). Thus, it is likely that catecholamine stimulation of membrane cAMP-dependent protein kinase will most likely be through activation of adenylate cyclase and increased intracellular cAMP concentration.…”

Section: Resultsmentioning

confidence: 99%

Catecholamine regulation of human erythrocyte membrane protein kinase.

Takeuchi¹,

Sonenberg²

1979

J. Clin. Invest.

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A B S T R A C T The effect of catecholamines on membrane-associated protein kinase in the mature human erythrocyte was investigated. Protein kinase activity was assayed after isolation of membranes from intact erythrocytes incubated with and without catecholamines. Activation of the enzyme is expressed as the ratio of the extent of phosphorylation of exogenous protein substrate in the absence to that in the presence of 2.5 ,M cyclic AMP (cAMP). The potent ,8-adrenergic agonist, (-)isoproterenol (2 ,uM), (-)epinephrine (10 ,uM) and (-)norepinephrine (10 uM) stimulated the cAMP-dependent protein kinase in membranes, 38±7%, 31±6%, and 30±6%, respectively. Maximal stimulation of membrane protein kinase by 10 ,uM (-)epinephrine was obtained _30 min after initiation of the incubation of erythrocytes with the hormone. The concentrations of (-)catecholamines that gave half-maximal stimulation of the membrane protein kinase were 0.17 ,M for isoproterenol, 0.35 ,M for epinephrine, and 0.63 ,uM for norepinephrine. The membrane protein kinase response to 6-adrenergic agonists was found to be stereospecific. The stimulation of membrane protein kinase by 10 ,uM (-)epinephrine was inhibited by the ,8-adrenergic antagonist, (-)propranolol with EC50 = 0.60 ,uM, and the inhibition of agonist stimulation of the cAMP-dependent protein kinase by propranolol was stereospecific. These studies suggest that a functional ,8-adrenergic receptor exists in the mature human erythrocyte.

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“…Since previous attempts to identify P-adrenergic receptors directly with radioactively labelled agonists were unsuccessful in brown adipose tissue [28] as in many other tissues [23,30], we used a potent P-adren- 24], in order to characterize such receptors in brown adipose tissue of warm-acclimated and cold-acclimated rats.…”

Section: Discussionmentioning

confidence: 99%

β‐Adrenergic Receptors in Brown‐Adipose Tissue. Characterization and Alteratios during Acclimation of Rats to Cold

Bukowiecki

Follea

Vallières

et al. 1978

European Journal of Biochemistry

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The capacity of brown adipose tissue to respond calorigenically to catecholamines increases markedly during cold‐acclimation of adult rats. To investigate this phenomenon, we have used the potent radioactive ligand (‐)‐[3H]dihydroalprenolol to directly estimate the number, the density and the affinity of β‐adrenergic receptors in brown adipose tissue membranes from control, cold‐exposed and cold‐acclimated rats. Binding of (−)‐[3H]dihydroalprenolol to unfractionated membranes was rapid, stable, saturable and reversible. It displayed the affinity, specificity and stereoselectivity expected of binding to adenylate cyclase‐coupled β‐adrenergic receptors. β‐Adrenergic agonists competed for binding sites with an order of potency typical of the β1 subtype of adrenergic receptors: (−)‐Isoproterenol > (−)‐norepinephrine ≥ (−)‐epinephrine. Binding exhibited a remarkable stereoselectivity, the (−)‐isomers of β‐adrenergic agonists and antagonists being 34 to 280 times more potent than the (+)‐isomers in competing for (−)‐[3H]dihydroalprenolol binding sites. Total interscapular brown adipose tissue of the adult, warm‐acclimated rat contained 1.12 ± 0.08 pmol of (−)‐[3H]dihydroalprenolol binding sites. During cold‐acclimation, growth of the tissue was accompanied by a 4–5‐fold increase in the total number of recptor sites. However, this increase did not keep pace with the increse in brown adipose tissue cellularity (as estimated by total tissue DNA content), resulting in a 40–50% reduction in receptor density. The decrease in receptor density was associated with cold‐exposure rather than with cold‐acclimation. The affinity of (−)‐[3H]dihydroalprenolol receptor sites was not significantly altered by cold‐acclimation. The results of this study are interpreted as indicating that catecholamines released via activation of the sympathetic nervous system regulate both the density and the number of their own receptors in brown adipose tissue of cold‐exposed animals. The development of the hyperadrenergic response of this thermogenic tissue during cold‐acclimation may result from a marked organ hyperplasia associated with an increased number of β‐adrenergic receptor sites and cannot be explained by alterations in receptor density or affinity.

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Problems in identification of the beta-adrenergic receptor.

Cited by 39 publications

References 0 publications

Isolation of adenylate cyclase-free, beta-adrenergic receptor from turkey erythrocyte membranes by affinity chromatography.

Isolation of adenylate cyclase-free, beta-adrenergic receptor from turkey erythrocyte membranes by affinity chromatography.

Catecholamine regulation of human erythrocyte membrane protein kinase.

β‐Adrenergic Receptors in Brown‐Adipose Tissue. Characterization and Alteratios during Acclimation of Rats to Cold

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