Ligand binding to the β-adrenergic receptor involves its rhodopsin-like core

Dixon, Richard A. F.; Sigal, Irving S.; Rands, Elaine; Register, R B; Candelore, Mari R.; Blake, Allan D.; Strader, Catherine D.

doi:10.1038/326073a0

Cited by 312 publications

(110 citation statements)

References 25 publications

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“…This suggestion would explain the ready solvation of transducin by aqueous solvents. The provocative proposal that interactions of membrane-spanning receptor (and adenylate cyclase) with G proteins take place outside the membrane in the aqueous phase is supported by recent data with a series of deletion mutants of the hamster fl,-adrenoceptor expressed in mammalian cells [54].…”

Section: Topographymentioning

confidence: 62%

Regulation of signal transfer from beta1-adrenoceptor to adenylate cyclase by betagamma subunits in a reconstituted system

et al. 1987

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The By subunits of guanine nucleotide binding proteins from bovine brain and bovine rod outer segments have different structural and immunochemical properties. In spite of these structural differences, py subunits from these sources have been found to be fully interchangeable in terms of their interaction with a subunits of pertussistoxin-sensitive G proteins. In contrast, however, there are striking differences between these by subunits with regard to their ability to deactivate fluoride-stimulated G,. These profound differences were also observed when the interaction of the purified components of the adenylate cyclase system was studied after reconstitution into phospholipid vesicles. Addition of by subunits purified from bovine brain to vesicles containing P-receptor and G, results in a biphasic effect on receptor-stimulated GTPase activity, whereas addition of transducin py was virtually without any effect. Likewise, by from bovine brain, but not transducin by, affected adenylate cyclase activity of a reconstituted system consisting of three purified components (R, G,, C). Thus, the a subunit of G,, but not the a subunits of pertussis-toxin-sensitive G proteins discriminate between structurally different Py subunits.It is now generally accepted that GTP-binding proteins play a universal role as receptor-effector couplers in hormonal and visual signal transduction. But whereas G, and Gi stimulate and inhibit adenylate cyclase, respectively, GI activates a retinal cGMP phosphodiesterase presumably by removing an inhibitory y subunit of the enzyme [l, 21. In addition, there are other G proteins, such as Go, G, and G, [3 -51. It is possible that these G proteins participate in other, not yet defined, receptor-effector coupling systems. All signaltransducing G proteins which have been characterized on a molecular basis have been found to be heterotrimers. The masses of the a subunits of the G proteins which bind and hydrolyze GTP range over 21 -52 kDa. All a subunits are substrates for ADP-ribosylation catalyzed by one or more bacterial toxin. Data obtained by molecular cloning have revealed that all a subunits studied so far differ structurally but that their homologies are great enough to consider them to belong to one class of related proteins [6]. The , ! 3 subunits of the G-protein heterotrimers isolated from different sources contain, with the exception of transducin, two chains with [8] indicate that the 35-kDa chain differs from the 36-kDa one. These data are corroborated by the existence of two genes for subunits [9]. Moreover, the y subunit of transducin seems to differ from all other known y subunits Until now, the information available is scarce on how the structural differences between the various by subunits may be related to functional differences. It was shown, for example, that transducin Py subunits and those from rabbit liver membranes can both support coupling of transducin a to lightactivated rhodopsin [12]. Moreover, it was shown by Cerione et al. [13] that transducin fly subunits can inhib...

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Section: Topographymentioning

confidence: 62%

Regulation of signal transfer from beta1-adrenoceptor to adenylate cyclase by betagamma subunits in a reconstituted system

et al. 1987

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“…Deletion mutagenesis and proteolytic cleavage studies of the ␤AR show that most of the connecting hydrophilic regions can be deleted without affecting ligand binding properties, suggesting that the ligand binding site is in the barrel of the TM region (48). Mutation of Asp-138 reduces the binding to both agonists and antagonists (16,17), suggesting that this residue is involved in binding and that agonists and antagonists might have similar binding sites.…”

Section: Resultsmentioning

confidence: 99%

Prediction of structure and function of G protein-coupled receptors

Vaidehi

Floriano

Trabanino

et al. 2002

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

278

258

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G protein-coupled receptors (GPCRs) mediate our sense of vision, smell, taste, and pain. They are also involved in cell recognition and communication processes, and hence have emerged as a prominent superfamily for drug targets. Unfortunately, the atomic-level structure is available for only one GPCR (bovine rhodopsin), making it difficult to use structure-based methods to design drugs and mutation experiments. We have recently developed first principles methods (MembStruk and HierDock) for predicting structure of GPCRs, and for predicting the ligand binding sites and relative binding affinities. Comparing to the one case with structural data, bovine rhodopsin, we find good accuracy in both the structure of the protein and of the bound ligand. We report here the application of MembStruk and HierDock to ␤1-adrenergic receptor, endothelial differential gene 6, mouse and rat I7 olfactory receptors, and human sweet receptor. We find that the predicted structure of ␤1-adrenergic receptor leads to a binding site for epinephrine that agrees well with the mutation experiments. Similarly the predicted binding sites and affinities for endothelial differential gene 6, mouse and rat I7 olfactory receptors, and human sweet receptor are consistent with the available experimental data. These predicted structures and binding sites allow the design of mutation experiments to validate and improve the structure and function prediction methods. As these structures are validated they can be used as targets for the design of new receptor-selective antagonists or agonists for GPCRs.GPCR ͉ olfactory receptor ͉ ␤-adrenergic receptor ͉ endothelial differentiation gene ͉ taste receptor G protein-coupled receptors (GPCRs) mediate senses such as odor, taste, vision, and pain (1) in mammals. In addition, important cell recognition and communication processes often involve GPCRs. Indeed, many diseases involve malfunction of these receptors (2), making them important targets for drug development. Unfortunately, despite their importance there is insufficient structural information on GPCRs for structure-based drug design. This is because these membrane-bound proteins are difficult to crystallize, and the atomic-level structure has been solved only for bovine rhodopsin (3, 4). Consequently, it is important to develop theoretical methods to predict the structure and function of GPCRs (5, 6).Experimental data relevant to the function of GPCRs is available for ligand activation of GPCRs (7-15) and site-directed mutagenesis (16)(17)(18). This data has led to information about structural features in the ligand-binding regions of GPCRs (refs. 5 and 19, and references therein). Protein sequence analyses on GPCRs reveals a common protein topology consisting of a membrane-spanning seven-helix bundle, which likely accommodates the binding site for low-molecular-weight ligands. Structurally, GPCRs can be classified as (i) GPCRs with short N terminus (5-80 residues) and (ii) GPCRs with a long N-terminal ectodomain (Ϸ80-600 residues). The long N terminus of ...

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“…In these experiments, a series of agonists with a variety of electron donating (i.e., CH 3 , OCH 3 , OH ranked in order of increasing electron- donating strength) and/or electron withdrawing (i.e., Cl) substituents in the catechol ring of the agonist were used. Since the strength of an aromatic interaction is dependent upon the magnitude of the charge separation in the ring, we hypothesized that these substituents would influence the dipole strength sufficiently such that the magnitude of the affinity increase or decrease would be altered accordingly with electron-donating groups increasing aromaticity and electron-withdrawing groups decreasing aromaticity.…”

Section: Resultsmentioning

confidence: 99%

“…Comparison of the amino acid sequences of the cloned adrenergic receptors illustrates the greatest residue conservation is found within the transmembrane helix domains. Accordingly, the binding pocket for epinephrine and norepinephrine is localized to this region in all adrenergic receptors, within the circular array of TM helices in the rhodopsin-like core of the receptor (3). Although differences in the agonist binding interactions at different adrenergic receptor subtypes exist, in general, the catecholamine is stabilized in the binding pocket by an ionic interaction involving the protonated amine of epinephrine and an aspartic acid residue in TM3 (4), by hydrogen bonds between the catechol hydroxyl groups of the agonist and serine residues in TM5 (5), and by an aromatic/hydrophobic interaction involving the catechol ring of the agonist and a phenylalanine residue in TM6 (6).…”

mentioning

confidence: 99%

Novel Aromatic Residues in Transmembrane Domains IV and V Involved in Agonist Binding at α1a-Adrenergic Receptors

Waugh

Zhao

Zuscik

et al. 2000

Journal of Biological Chemistry

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We examined the role that aromatic residues located in the transmembrane helices of the ␣ 1a -adrenergic receptor play in promoting antagonist binding. Since ␣ 1 -antagonists display low affinity binding at ␤ 2 -adrenergic receptors, two phenylalanine residues, Phe-163 and Phe-187, of the ␣ 1a -AR were mutated to the corresponding ␤ 2 -residue. Neither F163Q nor F187A mutations of the ␣ 1a had any effect on the affinity of the ␣ 1 -antagonists. However, the affinity of the endogenous agonist epinephrine was reduced 12.5-and 8-fold by the F163Q and F187A mutations, respectively. An additive loss in affinity (150-fold) for epinephrine was observed at an ␣ 1a containing both mutations. The loss of agonist affinity scenario could be reversed by a gain of affinity with mutation of the corresponding residues in the ␤ 2 to the phenylalanine residues in the ␣ 1a . We propose that both Phe-163 and Phe-187 are involved in independent aromatic interactions with the catechol ring of agonists. The potency but not the efficacy of epinephrine in stimulating phosphatidylinositol hydrolysis was reduced 35-fold at the F163Q/F187A ␣ 1a relative to the wild type receptor. Therefore, Phe-163 and Phe-187 represent novel binding contacts in the agonist binding pocket of the ␣ 1a -AR, but are not involved directly in receptor activation.The adrenergic receptors (␣ 1a , ␣ 1b , ␣ 1d , ␣ 2a , ␣ 2b , ␣ 2c , ␤ 1 , ␤ 2 , and ␤ 3 ) are part of a larger family of membrane proteins commonly referred to as the G protein-coupled receptor family. All of these AR subtypes bind the endogenous catecholamines, epinephrine and norepinephrine and mediate the actions of the sympathetic nervous system (1). The proposed topography adopted by these G protein-coupled receptors in the plasma membrane is modeled to that of bacteriorhodopsin, in which the protein folds to a highly ordered structure comprised of a series of seven hydrophobic transmembrane (TM) 1 -spanning domains, linked by three intracellular and three extracellular loops (2). Comparison of the amino acid sequences of the cloned adrenergic receptors illustrates the greatest residue conservation is found within the transmembrane helix domains. Accordingly, the binding pocket for epinephrine and norepinephrine is localized to this region in all adrenergic receptors, within the circular array of TM helices in the rhodopsin-like core of the receptor (3). Although differences in the agonist binding interactions at different adrenergic receptor subtypes exist, in general, the catecholamine is stabilized in the binding pocket by an ionic interaction involving the protonated amine of epinephrine and an aspartic acid residue in TM3 (4), by hydrogen bonds between the catechol hydroxyl groups of the agonist and serine residues in TM5 (5), and by an aromatic/hydrophobic interaction involving the catechol ring of the agonist and a phenylalanine residue in TM6 (6). The ␤-hydroxyl of the agonist, which confers stereoselectivity, has been identified in the ␤ 2 -AR as interacting with Asn-293 in TM6 (7), but is...

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Ligand binding to the β-adrenergic receptor involves its rhodopsin-like core

Cited by 312 publications

References 25 publications

Regulation of signal transfer from beta1-adrenoceptor to adenylate cyclase by betagamma subunits in a reconstituted system

Regulation of signal transfer from beta1-adrenoceptor to adenylate cyclase by betagamma subunits in a reconstituted system

Prediction of structure and function of G protein-coupled receptors

Novel Aromatic Residues in Transmembrane Domains IV and V Involved in Agonist Binding at α1a-Adrenergic Receptors

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