Transmembrane signaling by GPCRs: Insight from rhodopsin and opsin structures

Choe, Huhn; Park, Jung Hee; Kim, Yong Ju; Ernst, Oliver P.

doi:10.1016/j.neuropharm.2010.07.018

Cited by 49 publications

(49 citation statements)

References 54 publications

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“…Two recent lines of technologies and data have now contributed such visualization data to the field. The first is the elucidation of the structures of many GPCRs, encompassing both inactive and, more recently, active forms of the receptors, but also their downstream G-proteins Park et al, 2008;Scheerer et al, 2008;Rosenbaum et al, 2009Rosenbaum et al, , 2011Choe et al, 2011b). The second is the visualization of the receptors themselves as well as their signaling cascades by optical methods; these methods are based on the genetic or chemical labeling of GPCRs and their downstream signaling proteins and allow the monitoring of both their cellular localization and their activity (Kallal and Benovic, 2000;Boute et al, 2002;Lohse et al, 2008b;Balla, 2009).…”

Section: A G-protein-coupled Receptors and Methods Of Their Localizamentioning

confidence: 99%

Fluorescence/Bioluminescence Resonance Energy Transfer Techniques to Study G-Protein-Coupled Receptor Activation and Signaling

2012

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Section: A G-protein-coupled Receptors and Methods Of Their Localizamentioning

confidence: 99%

Fluorescence/Bioluminescence Resonance Energy Transfer Techniques to Study G-Protein-Coupled Receptor Activation and Signaling

2012

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“…Unlike rhodopsin, which exhibits an efficient and ordered photon-induced transition from the dark state conformation to metarhodopsin II (Choe et al, 2011a;Choe et al, 2011b), ligand-activated GPCRs generally show considerable basal activity that is reduced by inverse agonists.…”

Section: Introductionmentioning

confidence: 99%

Ligand Modulation of Sidechain Dynamics in a Wild-Type Human GPCR

Clark

Dikiy

Chapman

et al. 2018

Biophysical Journal

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GPCRs regulate all aspects of human physiology, and biophysical studies have deepened our understanding of GPCR conformational regulation by different ligands. Yet there is no experimental evidence for how sidechain dynamics control allosteric transitions between GPCR conformations. To address this deficit, we generated samples of a wild-type GPCR (A 2A R) that are deuterated apart from 1 H/ 13 C NMR probes at isoleucine d1 methyl groups, which facilitated 1 H/ 13 C methyl TROSY NMR measurements with opposing ligands. Our data indicate that low [Na + ] is required to allow large agonist-induced structural changes in A 2A R, and that patterns of sidechain dynamics substantially differ between agonist (NECA) and inverse agonist (ZM241385) bound receptors, with the inverse agonist suppressing fast ps-ns timescale motions at the G protein binding site. Our approach to GPCR NMR creates a framework for exploring how different regions of a receptor respond to different ligands or signaling proteins through modulation of fast ps-ns sidechain dynamics.

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“…Despite their abundance and functional importance, membrane proteins are largely underrepresented in the Protein Data Bank, mainly due to technical difficulties in studying these proteins. A large fraction of these proteins such as G protein-coupled receptors (GPCRs) transduces signals across membranes through conformational changes (3)(4)(5)(6). The resulting protein conformational heterogeneity and flexibility is a major factor hindering crystallization.…”

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confidence: 99%

Naturally evolved G protein-coupled receptors adopt metastable conformations

Chen

Zhou

Fryszczyn

et al. 2012

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

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A wide range of membrane receptors signal through conformational changes, and the resulting protein conformational flexibility often hinders their structural studies. Because the determinants of membrane receptor conformational stability are still poorly understood, identifying a minimal set of perturbations stabilizing a membrane protein in a given conformation remains a major challenge in membrane protein structure determination. We present a novel approach integrating bioinformatics, computational design and experimental techniques that identifies and stabilizes metastable receptor regions. When applied to the beta1-adrenergic receptor, the method generated 13 novel receptor variants stabilized in the intended inactive state among which two exhibit an apparent thermostability higher than WT and M23 (a receptor variant previously stabilized by extensive scanning mutagenesis) by more than 30°C and 11°C, respectively. Targeted regions involve nonconserved unsatisfied polar residues or exhibit significant packing defects, features found in all class A G protein-coupled receptor structures. These findings suggest that natural G protein-coupled receptor sequences have evolved to be conformationally metastable through the design of suboptimal polar and van der Waals tertiary interactions. Given sufficiently accurate structural models, our approach should prove useful for designing stabilized variants of many uncharacterized membrane receptors.computational protein design | protein conformational stability | membrane protein modeling | signal transduction | synthetic biology M embrane proteins represent around 30% of currently sequenced genomes and are critical in the regulation of cell signaling and cell-cell communication (1, 2). When dysfunctional, however, these proteins can be responsible for serious diseases and constitute more than 60% of current drug targets. Despite their abundance and functional importance, membrane proteins are largely underrepresented in the Protein Data Bank, mainly due to technical difficulties in studying these proteins. A large fraction of these proteins such as G protein-coupled receptors (GPCRs) transduces signals across membranes through conformational changes (3-6). The resulting protein conformational heterogeneity and flexibility is a major factor hindering crystallization. An additional obstacle to structural determination is the poor stability of membrane proteins in detergent solution required by traditional crystallization approaches (7,8).In recent years, several approaches to stabilize membrane proteins have been explored (9). These methods include development and optimization of solubilizing detergents (10), reconstitution of proteins into lipidic bicelles (11, 12), cleavage of flexible protein regions (13-18), co-crystallization with stabilizing ligands, antibodies, and fusion with soluble proteins (e.g., T4 lysozyme) (13-15, 17, 19-23). Several of these modifications, however, disrupt important functional regions and interactions with either natural ligands or downstre...

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Transmembrane signaling by GPCRs: Insight from rhodopsin and opsin structures

Cited by 49 publications

References 54 publications

Fluorescence/Bioluminescence Resonance Energy Transfer Techniques to Study G-Protein-Coupled Receptor Activation and Signaling

Fluorescence/Bioluminescence Resonance Energy Transfer Techniques to Study G-Protein-Coupled Receptor Activation and Signaling

Ligand Modulation of Sidechain Dynamics in a Wild-Type Human GPCR

Naturally evolved G protein-coupled receptors adopt metastable conformations

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