Role of the β-Adrenergic Receptor in the Regulation of Adenylate Cyclase

Smigel, Murray D.; Ross, Elliott M.; Gilman, Alfred G.

doi:10.1007/978-1-4684-4823-8_7

Cited by 19 publications

(13 citation statements)

References 164 publications

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“…$49 murine lymphoma cells which possess ~-adrenergic receptors coupled to adenylyl cyclase via Gs (see [33]) were used to examine the functional consequences of RM treatment. Isoproterenol-dependent activation of Gs mediated by the fl-adrenergic receptor was inhibited by RM in a concentration-dependent fashion in membranes of wild-type $49 murine lymphoma cells with an ICs0 value of approx.…”

Section: Resultsmentioning

confidence: 99%

Receptor and effector interactions of G_s Functional studies with antibodies to the α_s carboxyl‐terminal decapeptide

et al. 1989

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Antibodies generated to a synthetic decapeptide, RMHLRQYELL, representing the carboxyl-terminus of Gs-ct have been characterized in immunoblots and functional studies. This antibody, designated RM, reacts exclusively with a doublet of proteins of 52 and 45 kDa in immunoblots of bovine brain and wild-type $49 murine lymphoma cell membranes. No such reactivity is seen in membranes from cyc-$49 cells, which lack Gs. RM blocks receptor-mediated activation of Gs and adenylyl cyclase in membranes from wild-type $49 cells. RM could also immunoprecipitate adenylyl cyclase activity in detergent extracts from GTP[),]S-or fluoride-preactivated bovine brain membranes; thus binding of cts to effector and carboxyl-terminal antibody was mutually compatible. Such experiments provide an approach for the elucidation of functionally relevant interactions of G-proteins with receptors and effectors in the membrane.

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Section: Resultsmentioning

confidence: 99%

Receptor and effector interactions of G_s Functional studies with antibodies to the α_s carboxyl‐terminal decapeptide

et al. 1989

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“…Because both proteins act by regulating GTP-binding proteins, a compact structure based on seven membrane-spanning regions may be a general model for receptors that act on G proteins. (1). Functionally, the receptor is phylogenetically conserved.…”

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confidence: 99%

“…Activated G, can then bind to adenylate cyclase and stimulate its activity (see ref. 1 for review). The ,B-adrenergic receptor is thus a member of an even larger class of cell surface receptors that regulate the functions of multiple GTP-binding regulatory proteins, or G proteins.…”

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confidence: 99%

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The avian beta-adrenergic receptor: primary structure and membrane topology.

Yarden¹,

Rodriguez²,

Wong³

et al. 1986

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

223

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Partial amino acid sequence information allowed the isolation of cDNA clones encoding the turkey erythrocyte (3-adrenergic receptor. Antisera raised against synthetic peptides encoded by the cDNA crossreacted with the purified receptor and appropriate tryptic fragments, confirming the identity of the cDNA. The receptor is composed of 483 amino acids and has a molecular mass of 54 kDa. Its sequence suggests that it is arranged predominantly in seven membranespanning sequences and a long cytoplasmic carboxyl-terminal domain. The extracellular amino-terminal domain contains a consensus sequence for N-glycosylation. The /3-adrenergic receptor displays overall structural similarity and weak sequence homology with rhodopsin. Because both proteins act by regulating GTP-binding proteins, a compact structure based on seven membrane-spanning regions may be a general model for receptors that act on G proteins. (1). Functionally, the receptor is phylogenetically conserved. The receptor purified from turkey erythrocytes can efficiently regulate GO from rabbit liver in reconstituted phospholipid vesicles (3, 4), and its selectivity for numerous agonist and antagonist ligands is only slightly discrepant from the mammalian 31-adrenergic receptor (5).Whether the receptor interacts with G. on the hydrophilic cytoplasmic face of the plasma membrane or within the bilayer is unknown; nor is anything known about the structural details of this regulatory interaction. Presumably, all receptors that activate G proteins will share this regulatory domain, and definable differences will exist in the homologous regions of receptors that activate different G proteins.The sites of P-adrenergic ligand binding, regulatory phosphorylation, and stimulatory reduction by thiols (6) are also of great interest. Much of the difficulty in learning about the structure of the f3-adrenergic receptor is due to its low concentration in plasma membranes: P-adrenergic receptor must be purified over 20,000-fold from an already wellpurified plasma membrane fraction. We have therefore undertaken the cloning of the cDNA that encodes the Padrenergic receptor as a first step toward more direct studies of its structure and function. The sequence of the 82-adrenergic receptor from hamster lung has appeared recently (7), and homology between the two sequences and the sequence of rhodopsin suggests functionally important aspects of their structures. METHODSP3-Adrenergic receptor was purified from turkey erythrocyte plasma membrane as described by Brandt and Ross (4). This preparation, which consists mainly of an active 40-kDa proteolysis product and the 53-kDa receptor (1, 4), was separated from digitonin and minor impurities by HPLC on a 300-A pore size, C4 column (Synchrom, Linden, IN) using a linear gradient of0.1% trifluoroacetic acid in water to 0.06% trifluoroacetic acid in 50% 1-propanol (vol/vol). The receptor was cleaved with cyanogen bromide, 60 mg/ml in 70% (vol/vol) formic acid, for 24 hr at room temperature under nitrogen. Sequencing was performed...

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“…*3-;='GTPyS * Gw + (3 Mg2+ Gi,0. (+ GTPyS GTPyS QGa j *' GTPyS 0* Gi+ (3 The obvious next question was to determine which of the subunits was responsible for the characteristic effects ofthe "activated" dimer. Resolution of the subunits can be achieved by high-performance gel filtration, and it was then determined that GTPzyS -G. was the necessary and sufficient activator of C. The (-subunit of G, thus acts as an inhibitor of activation and as a stimulator of deactivation by promoting formation of the G, dimer.…”

mentioning

confidence: 99%

Guanine nucleotide-binding regulatory proteins and dual control of adenylate cyclase.

Gilman¹

1984

J. Clin. Invest.

334

126

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Role of the β-Adrenergic Receptor in the Regulation of Adenylate Cyclase

Cited by 19 publications

References 164 publications

Receptor and effector interactions of G_s Functional studies with antibodies to the α_s carboxyl‐terminal decapeptide

Receptor and effector interactions of G_s Functional studies with antibodies to the α_s carboxyl‐terminal decapeptide

The avian beta-adrenergic receptor: primary structure and membrane topology.

Guanine nucleotide-binding regulatory proteins and dual control of adenylate cyclase.

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Role of the β-Adrenergic Receptor in the Regulation of Adenylate Cyclase

Cited by 19 publications

References 164 publications

Receptor and effector interactions of Gs Functional studies with antibodies to the αs carboxyl‐terminal decapeptide

Receptor and effector interactions of Gs Functional studies with antibodies to the αs carboxyl‐terminal decapeptide

The avian beta-adrenergic receptor: primary structure and membrane topology.

Guanine nucleotide-binding regulatory proteins and dual control of adenylate cyclase.

Contact Info

Product

Resources

About

Receptor and effector interactions of G_s Functional studies with antibodies to the α_s carboxyl‐terminal decapeptide

Receptor and effector interactions of G_s Functional studies with antibodies to the α_s carboxyl‐terminal decapeptide