The structure and function of G-protein-coupled receptors

Rosenbaum, Daniel M.; Rasmussen, Søren G. F.; Kobilka, Brian K.

doi:10.1038/nature08144

Cited by 2,100 publications

(1,806 citation statements)

References 67 publications

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“…As a model of GPCRs, Rho has been extensively studied, and since the first report of its crystal structure, several GPCRs structures have been solved by X-ray crystallography [1][2][3][4][5] . Rho is found in the rod outer segment (ROS) membranes of the photoreceptor cells of the retina where it is embedded in the lipid bilayer surrounded by ~65-70 phospholipids per protein molecule [6][7][8][9] .…”

Section: Introductionmentioning

confidence: 99%

Phospholipid Bicelles Improve the Conformational Stability of Rhodopsin Mutants Associated with Retinitis Pigmentosa

et al. 2015

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Mutations in the visual photoreceptor rhodopsin are the cause of the retinal degenerative disease retinitis pigmentosa. Some naturally occurring mutations can lead to protein conformational instability. Two such mutations, N55K and G90V, in the first and second transmembrane helices of the receptor, have been associated with sector and classical retinitis pigmentosa, respectively, and showed enhanced thermal sensitivity. We have carefully analyzed the effect of phospholipid bicelles on the stability and ligand binding properties of these two mutants and compared it with those of the detergent-solubilized samples. We have used a phospholipid bilayer consisting of 1,2-dimyristoyl-sn-glycero-3-phosphatidylcholine (DMPC) and 1,2dihexanoyl-sn-glycero-3-phosphocholine (DHPC). We find that DMPC/DHPC bicelles dramatically increase the thermal stability of the rhodopsin mutants G90V and N55K. The chromophore stability and regeneration of the mutants were also increased in bicelles when compared to their behavior in a dodecyl maltoside detergent solution. The retinal release process was slowed in bicelles, and chromophore entry, after illumination, was improved for the G90V mutant but not for N55K. Furthermore, fluorescence spectroscopy measurements showed that bicelles allowed more exogenous retinal binding to the photoactivated G90V mutant than in a detergent solution. In contrast, N55K could not reposition any chromophore either in the detergent or in bicelles. The results demonstrate that DMPC/DHPC bicelles can counteract the destabilizing effect of the disease-causing mutations and can modulate the structural changes in the ensuing receptor photoactivation in a distinct specific manner for different retinitis pigmentosa mutant phenotypes.

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Section: Introductionmentioning

confidence: 99%

Phospholipid Bicelles Improve the Conformational Stability of Rhodopsin Mutants Associated with Retinitis Pigmentosa

et al. 2015

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“…An early discrimination of the investigated ligands into these categories in silico is of high importance for pharmaceutical research. Most GPCR crystal structures represent an inactive state which has been considered to be capable for the discovery of antagonists and inverse agonists only [12][13][14]. To date, only the structure of opsin [15] and most recently structural models of active conformations of the b 2 -adrenergic receptor (B2AR) [16][17][18], the agonist-bound b 1 -adrenergic receptor [19] and a constitutively active rhodopsin [20] could be potentially used as a template for the modelling of active GPCRs and subsequent virtual screening for agonists.…”

Section: Introductionmentioning

confidence: 99%

The structure of active opsin as a basis for identification of GPCR agonists by dynamic homology modelling and virtual screening assays

et al. 2011

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a b s t r a c tMost of the currently available G protein-coupled receptor (GPCR) crystal structures represent an inactive receptor state, which has been considered to be suitable only for the discovery of antagonists and inverse agonists in structure-based computational ligand screening. Using the b 2 -adrenergic receptor (B2AR) as a model system, we show that a dynamic homology model based on an ''active'' opsin structure without further incorporation of experimental data performs better than the crystal structure of the inactive B2AR in finding agonists over antagonists/inverse agonists. Such ''active-like state'' dynamic homology models can therefore be used to selectively identify GPCR agonists in in silico ligand libraries.

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“…They all activate heterotrimeric G proteins upon agonist binding, but their seven helices exhibit significant divergence, reflecting a high degree of ligand variation, from small amines to peptide hormones 9, 10 . A recent study demonstrated that, despite such variation among GPCRs, some conserved features in terms of intramolecular atomic distances were discernible 11 .…”

Section: Introductionmentioning

confidence: 99%

Sequence and intramolecular distance scoring analyses of microbial rhodopsins

Asano¹,

Ide²,

Kamata³

et al. 2016

F1000Res

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Recent accumulation of sequence and structural data, in conjunction with systematical classification into a set of families, has significantly advanced our understanding of diverse and specific protein functions. Analysis and interpretation of protein family data requires comprehensive sequence and structural alignments. Here, we present a simple scheme for analyzing a set of experimental structures of a given protein or family of proteins, using microbial rhodopsins as an example. For a data set comprised of around a dozen highly similar structures to each other (overall pairwise root-mean-squared deviation < 2.3 Å), intramolecular distance scoring analysis yielded valuable information with respect to structural properties, such as differences in the relative variability of transmembrane helices. Furthermore, a comparison with recent results for G protein-coupled receptors demonstrates how the results of the present analysis can be interpreted and effectively utilized for structural characterization of diverse protein families in general.

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The structure and function of G-protein-coupled receptors

Cited by 2,100 publications

References 67 publications

Phospholipid Bicelles Improve the Conformational Stability of Rhodopsin Mutants Associated with Retinitis Pigmentosa

Phospholipid Bicelles Improve the Conformational Stability of Rhodopsin Mutants Associated with Retinitis Pigmentosa

The structure of active opsin as a basis for identification of GPCR agonists by dynamic homology modelling and virtual screening assays

Sequence and intramolecular distance scoring analyses of microbial rhodopsins

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