Seven-transmembrane receptors: crystals clarify

Lu, Zhi-Liang; Saldanha, José W.; Hulme, Edward C.

doi:10.1016/s0165-6147(00)01973-8

Cited by 167 publications

(139 citation statements)

References 75 publications

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“…W6.48 is highly conserved throughout the class A GPCRs and has been proposed to be in close proximity to the agonist-binding site and to play a central role in receptor activation (9,33). In particular, binding-induced changes in the rotameric state of W6.48 are thought to act as part of a ''switch'' that is critical to receptor function (4,9,(33)(34)(35)(36)(37). W7.40 is the next most highly conserved residue associated with the aminergic class of GPCRs (38).…”

Section: Discussionmentioning

confidence: 99%

“…The joint interaction of the OCH 2 ONH 3 ϩ group with helices 3 and 6 agrees with the contemporary model for receptor activation that has the extracellular portion of helix 6 moving toward helix 3. This movement of helix 6 is part of a rotamer switch activation mechanism (4,9,(33)(34)(35)(36)(37). A key component of this model is a reorientation of the side chain of indole of W6.48 from a perpendicular to a parallel orientation, relative to the plane of the membrane, as the receptor transitions from the inactive to active state.…”

Section: Discussionmentioning

confidence: 99%

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Probing the role of the cation–π interaction in the binding sites of GPCRs using unnatural amino acids

Torrice¹,

Bower²,

Lester

et al. 2009

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

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We describe a general application of the nonsense suppression methodology for unnatural amino acid incorporation to probe drugreceptor interactions in functional G protein-coupled receptors (GPCRs), evaluating the binding sites of both the M2 muscarinic acetylcholine receptor and the D2 dopamine receptor. Receptors were expressed in Xenopus oocytes, and activation of a G protein-coupled, inward-rectifying K ؉ channel (GIRK) provided, after optimization of conditions, a quantitative readout of receptor function. A number of aromatic amino acids thought to be near the agonist-binding site were evaluated. Incorporation of a series of fluorinated tryptophan derivatives at W6.48 of the D2 receptor establishes a cation-interaction between the agonist dopamine and W6.48, suggesting a reorientation of W6.48 on agonist binding, consistent with proposed ''rotamer switch'' models. Interestingly, no comparable cationinteraction was found at the aligning residue in the M2 receptor.D2 receptor ͉ fluorination ͉ membrane protein G protein-coupled receptors (GPCRs) represent the largest family of transmembrane receptor proteins in the human genome and constitute a prominent class of targets for the pharmaceutical industry (1-3). Accordingly, they have been studied extensively throughout academia and industry, by using the full range of chemical, biochemical, and biophysical techniques. In recent years, the field has been energized by several high-resolution crystal structures of mammalian GPCRs that build upon the earlier, highly informative structural studies of rhodopsin and bacteriorhodopsin (4-9).The structural snapshots provided by crystallography greatly enhance our understanding of specific receptors but also raise many new issues. Key among these is the extent to which the information from available structures can be extrapolated to the hundreds of other GPCRs. In addition, a key goal in the study of GPCRs-and all receptors-is a description of the interconversions among several structural states that underlie the protein's biological function. It can be a challenging task to deduce a signaling mechanism from static images. As such, structure-function studies, guided by the new structural advances, will remain an important tool in evaluating GPCR function and the nature of drug-receptor interactions in this family.In recent years, unnatural amino acid mutagenesis on ion channels and receptors expressed in Xenopus oocytes has provided a powerful tool for uncovering crucial drug-receptor interactions and signaling mechanisms (10, 11). GPCRs present an especially attractive target for unnatural amino acid mutagenesis, given the importance of the family, the significant pharmacological variations among closely related family members, and the central role of structural rearrangements in their biological function.Incorporating unnatural amino acids into GPCRs, however, presents unique challenges. Most unnatural amino acid mutagenesis studies in eukaryotic cells have focused on ion channels. These studies exploit the exquisite s...

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Probing the role of the cation–π interaction in the binding sites of GPCRs using unnatural amino acids

Torrice¹,

Bower²,

Lester

et al. 2009

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

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“…The homology model of the M 1 muscarinic acetylcholine receptor has shown to be a good template for interpreting the structureactivity, mutagenesis and affinity labeling data available for this kind of receptors [18]. In addition, virtual screening of binding sites on homology models based on the X-ray structure of rhodopsin, proved that this method is suitable in several cases [19].…”

Section: Discussionmentioning

confidence: 99%

Identification of multiple allosteric sites on the M₁ muscarinic acetylcholine receptor

Espinoza-Fonseca

Trujillo‐Ferrara

2005

FEBS Letters

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Staurosporine and four staurosporine derivatives were docked on the rhodopsin-based homology model of the M 1 muscarinic acetylcholine receptor in order to localize the possible allosteric sites of this receptor. It was found that there were three major allosteric sites, two of which are located at the extracellular face of the receptor, and one in the intracellular domain of the receptor. In the present study, the localization of these binding sites is described for the first time. The present study confirms the existence of multiple allosteric sites on the M 1 muscarinic receptor, and lays the ground for further experimental and computational analysis to better understand how muscarinic receptors are modulated via their allosteric sites. These findings will also help to design and develop novel drugs acting as allosteric modulators of the M 1 receptor, which can be used in the treatment of the AlzheimerÕs disease.

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“…We prefer a reduction in intrinsic receptor efficacy after Cys 337 mutation, as demonstrated in experiments which analysed the decreased coupling of 'nonpalmitoylated' GPCRs (the M 2 muscarinic and endothelin receptor mutants) in reconstituted systems in the absence of kinase (Hayashi & Haga, 1997;Doi et al, 1999). In rhodopsin, the fourth intracellular loop forms a short helix (H8; Palczewski et al, 2000;Lu et al, 2002) which when disrupted by chemical depalmitoylation reduces signalling efficacy (Sachs et al, 2000). The conserved nature of this structural motif among GPCRs may therefore be a key to alterations in intrinsic efficacy associated with changes in the palmitoylation state.…”

Section: Discussionmentioning

confidence: 99%

Control of signalling efficacy by palmitoylation of the rat Y₁ receptor

Holliday

Cox

2003

British J Pharmacology

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Seven-transmembrane receptors: crystals clarify

Cited by 167 publications

References 75 publications

Probing the role of the cation–π interaction in the binding sites of GPCRs using unnatural amino acids

Probing the role of the cation–π interaction in the binding sites of GPCRs using unnatural amino acids

Identification of multiple allosteric sites on the M₁ muscarinic acetylcholine receptor

Control of signalling efficacy by palmitoylation of the rat Y₁ receptor

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Seven-transmembrane receptors: crystals clarify

Cited by 167 publications

References 75 publications

Probing the role of the cation–π interaction in the binding sites of GPCRs using unnatural amino acids

Probing the role of the cation–π interaction in the binding sites of GPCRs using unnatural amino acids

Identification of multiple allosteric sites on the M1 muscarinic acetylcholine receptor

Control of signalling efficacy by palmitoylation of the rat Y1 receptor

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Identification of multiple allosteric sites on the M₁ muscarinic acetylcholine receptor

Control of signalling efficacy by palmitoylation of the rat Y₁ receptor