G protein-coupled receptors: the evolution of structural insight

Gacasan, Samantha B.; Baker, Daniel L.; Parrill, Abby L.

doi:10.3934/biophy.2017.3.491

Cited by 63 publications

(65 citation statements)

References 189 publications

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“…The majority of studies on isolated membrane proteins are still carried out in a detergent system; GPCR research is no exception ( Table 2 ) where well-established protocols have greatly simplified membrane extraction. The vast majority of GPCR structures deposited in the PDB have been obtained for receptors in detergent micelles [ 185 ]. Further, the proven ability to make homogeneous particles with sizes ( Figure 4 ) that relate to sufficiently rapid rotational tumbling means detergents have been a favoured system for solution NMR studies.…”

Section: Membrane Mimetic Systems For Structural and Functional Stmentioning

confidence: 99%

Structure and Dynamics of GPCRs in Lipid Membranes: Physical Principles and Experimental Approaches

et al. 2020

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Over the past decade, the vast amount of information generated through structural and biophysical studies of GPCRs has provided unprecedented mechanistic insight into the complex signalling behaviour of these receptors. With this recent information surge, it has also become increasingly apparent that in order to reproduce the various effects that lipids and membranes exert on the biological function for these allosteric receptors, in vitro studies of GPCRs need to be conducted under conditions that adequately approximate the native lipid bilayer environment. In the first part of this review, we assess some of the more general effects that a membrane environment exerts on lipid bilayer-embedded proteins such as GPCRs. This is then followed by the consideration of more specific effects, including stoichiometric interactions with specific lipid subtypes. In the final section, we survey a range of different membrane mimetics that are currently used for in vitro studies, with a focus on NMR applications.

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Section: Membrane Mimetic Systems For Structural and Functional Stmentioning

confidence: 99%

Structure and Dynamics of GPCRs in Lipid Membranes: Physical Principles and Experimental Approaches

et al. 2020

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“…The domain orientations are constrained by the mutational data in such a way as to bring ECL2 immediately below the last module of the CRD. In GPCRs generally, ECL2 is a critical determinant of ligand interactions (55,56). ECL2 of class C GPCRs are similar in length (less than 35 residues), and the length of the linker peptide at the end of the CRD is also important for receptor activation and surface expression (57)(58)(59).…”

Section: Structural Architecture Of the Dimeric Sweet Taste Receptormentioning

confidence: 99%

Structural architecture of a dimeric class C GPCR based on co-trafficking of sweet taste receptor subunits

Park

Selvam

Sanematsu

et al. 2019

Journal of Biological Chemistry

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Edited by Henrik G. Dohlman Class C G protein-coupled receptors (GPCRs) are obligatory dimers that are particularly important for neuronal responses to endogenous and environmental stimuli. Ligand recognition through large extracellular domains leads to the reorganization of transmembrane regions to activate G protein signaling. Although structures of individual domains are known, the complete architecture of a class C GPCR and the mechanism of interdomain coupling during receptor activation are unclear. By screening a mutagenesis library of the human class C sweet taste receptor subunit T1R2, we enhanced surface expression and identified a dibasic intracellular retention motif that modulates surface expression and co-trafficking with its heterodimeric partner T1R3. Using a highly expressed T1R2 variant, dimerization sites along the entire subunit within all the structural domains were identified by a comprehensive mutational scan for co-trafficking with T1R3 in human cells. The data further reveal that the C terminus of the extracellular cysteine-rich domain needs to be properly folded for T1R3 dimerization and co-trafficking, but not for surface expression of T1R2 alone. These results guided the modeling of the T1R2-T1R3 dimer in living cells, which predicts a twisted arrangement of domains around the central axis, and a continuous folded structure between transmembrane domain loops and the cysteine-rich domains. These insights have implications for how conformational changes between domains are coupled within class C GPCRs. This work was supported in part by National Institutes of Health NIMH Award R21MH113155 (to E. P.) for development of deep mutational scanning in human cells and in part by JSPS KAKENHI Grant JP18K09523 (to K. S.). The authors declare that they have no conflicts of interest with the contents of this article. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. This article contains Figs. S1-S12 and Tables S1-S4. Data were deposited in NCBI's Gene Expression Omnibus under accession number GSE115751.

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“…This GPR17 model resulted much more accurate than the previous ones, due to the closest sequence identity of GPR17 with respect to P2Y 1 , compared to that shared with all the other available crystallographic templates among related class-A GPCRs (26.4%). A pairwise alignment identity matrix between GPR17 and the primary structures of other related class-A GPCR, whose structure has been solved (Gacasan et al, 2017), is reported in Supplementary Figure S5.…”

Section: Molecular Modeling and Ligand Dockingmentioning

confidence: 99%

Surface Plasmon Resonance as a Tool for Ligand Binding Investigation of Engineered GPR17 Receptor, a G Protein Coupled Receptor Involved in Myelination

et al. 2020

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The aim of this study was to investigate the potential of surface plasmon resonance (SPR) spectroscopy for the measurement of real-time ligand-binding affinities and kinetic parameters for GPR17, a G protein-coupled receptor (GPCR) of major interest in medicinal chemistry as potential target in demyelinating diseases. The receptor was directly captured, in a single-step, from solubilized membrane extracts on the sensor chip through a covalently bound anti-6x-His-antibody and retained its ligand binding activity for over 24 h. Furthermore, our experimental setup made possible, after a mild regeneration step, to remove the bound receptor without damaging the antibody, and thus to reuse many times the same chip. Two engineered variants of GPR17, designed for crystallographic studies, were expressed in insect cells, extracted from crude membranes and analyzed for their binding with two high affinity ligands: the antagonist Cangrelor and the agonist Asinex 1. The calculated kinetic parameters and binding constants of ligands were in good agreement with those reported from activity assays and highlighted a possible functional role of the N-terminal residues of the receptor in ligand recognition and binding. Validation of SPR results was obtained by docking and molecular dynamics of GPR17-ligands interactions and by functional in vitro studies. The latter allowed us to confirm that Asinex 1 behaves as GPR17 receptor agonist, inhibits forskolin-stimulated adenylyl cyclase pathway and promotes oligodendrocyte precursor cell maturation and myelinating ability.

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G protein-coupled receptors: the evolution of structural insight

Cited by 63 publications

References 189 publications

Structure and Dynamics of GPCRs in Lipid Membranes: Physical Principles and Experimental Approaches

Structure and Dynamics of GPCRs in Lipid Membranes: Physical Principles and Experimental Approaches

Structural architecture of a dimeric class C GPCR based on co-trafficking of sweet taste receptor subunits

Surface Plasmon Resonance as a Tool for Ligand Binding Investigation of Engineered GPR17 Receptor, a G Protein Coupled Receptor Involved in Myelination

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