Analysis of Human Dopamine D3 Receptor Quaternary Structure

Marsango, Sara; Caltabiano, Gianluigi; Pou, Chantévy; Liste, María José Varela; Milligan, Graeme

doi:10.1074/jbc.m114.630681

Cited by 23 publications

(29 citation statements)

References 54 publications

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“…2 ) as well as by recent experimental data for a close homolog of the D 2 receptor, namely the D 3 receptor dimer [ 42 ]. Marsango et al [ 42 ] used molecular modeling, mutagenesis and analysis of inactive state receptor crystal structures to indicate that D 3 monomers can interact with each other via at least two distinct interfaces: the first comprising residues from transmembrane domains TM1 and TM2 along with those from TM7, and a second involving transmembrane domains TM4 and TM5. Moreover, Guitart et al [ 43 ] also reported that the D 1 receptor TM5 peptide was also able to reduce D 1 –D 1 receptor complementation.…”

Section: Resultsmentioning

confidence: 95%

The dopamine D2 receptor dimer and its interaction with homobivalent antagonists: homology modeling, docking and molecular dynamics

2016

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In order to apply structure-based drug design techniques to G protein-coupled receptor complexes, it is essential to model their 3D structure and to identify regions that are suitable for selective drug binding. For this purpose, we have developed and tested a multi-component protocol to model the inactive conformation of the dopamine D2 receptor dimer, suitable for interaction with homobivalent antagonists. Our approach was based on protein–protein docking, applying the Rosetta software to obtain populations of dimers as present in membranes with all the main possible interfaces. Consensus scoring based on the values and frequencies of best interfaces regarding four scoring parameters, Rosetta interface score, interface area, free energy of binding and energy of hydrogen bond interactions indicated that the best scored dimer model possesses a TM4–TM5–TM7–TM1 interface, which is in agreement with experimental data. This model was used to study interactions of the previously published dopamine D2 receptor homobivalent antagonists based on clozapine,1,4-disubstituted aromatic piperidines/piperazines and arylamidoalkyl substituted phenylpiperazine pharmacophores. It was found that the homobivalent antagonists stabilize the receptor-inactive conformation by maintaining the ionic lock interaction, and change the dimer interface by disrupting a set of hydrogen bonds and maintaining water- and ligand-mediated hydrogen bonds in the extracellular and intracellular part of the interface.Graphical AbstractStructure of the final model of the dopamine D2 receptor homodimer, indicating the distancebetween Tyr37 and Tyr 5.42 in the apo form (left) and in the complex with the ligand (right).Electronic supplementary materialThe online version of this article (doi:10.1007/s00894-016-3065-2) contains supplementary material, which is available to authorized users.

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Section: Resultsmentioning

confidence: 95%

The dopamine D2 receptor dimer and its interaction with homobivalent antagonists: homology modeling, docking and molecular dynamics

2016

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“…However, in many of these studies, the point mutants did not eliminate energy transfer completely, which was interpreted as evidence of higher-order oligomers and multiple dimerization interfaces. 26–30 The difficulty with BRET and FRET studies is that they can be overly sensitive to weak and transient interactions and potentially overestimate the extent of dimerization. This interpretation was supported by data from several assays that are sensitive to dimer lifetimes.…”

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confidence: 99%

A G Protein-Coupled Receptor Dimerization Interface in Human Cone Opsins

et al. 2016

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G protein-coupled receptors (GPCRs) detect a wide variety of physical and chemical signals and transmit that information across the cellular plasma membrane. Dimerization is a proposed modulator of GPCR signaling, but the structure and stability of class A GPCR dimerization have been difficult to establish. Here we investigated the dimerization affinity and binding interface of human cone opsins, which initiate and sustain daytime color vision. Using a time-resolved fluorescence approach, we found that human red cone opsin exhibits a strong propensity for dimerization, whereas the green and blue cone opsins do not. Through mutagenesis experiments, we identified a dimerization interface in the fifth transmembrane helix of human red cone opsin involving amino acids I230, A233, and M236. Insights into this dimerization interface of red cone opsin should aid ongoing investigations of the structure and function of GPCR quaternary interactions in cell signaling. Finally, we demonstrated that the same residues needed for dimerization are also partially responsible for the spectral tuning of red cone opsin. This last observation has the potential to open up new lines of inquiry regarding the functional role of dimerization for red cone opsin.

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“…Different strategies have been used to map the GPCR dimer interface and its functional significance. These strategies include: (i) structure-based molecular modeling to predict residues potentially involved in dimer formation (42)(43)(44); (ii) cysteine mutagenesis of amino acids predicted to be involved in formation of the dimer followed by cross-linking, which successfully mapped several amino acids at the dimer interface of opsin, dopamine D2, and serotonin 5HT2c receptors (45)(46)(47); and (iii) disruption of the dimer interface by mutation of certain dimer-contacting residues (48,49) or with synthetic peptides derived from selected TM domains, resulting in specific alterations of receptor function, which for example was demonstrated for CXCR4 and formyl peptide receptors targeting TM4 (50,51) and for ␤ 2 AR and leukotriene B4 (BLT1) receptors targeting TM6 (52,53). However, TM peptides corresponding to TM6 and TM7 of the cholecystokinin receptor inhibited receptor dimerization without altering its function, as assessed by ligand binding and agonist-stimulated intracellular calcium concentration assays (25).…”

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confidence: 99%

Disruption of Rhodopsin Dimerization with Synthetic Peptides Targeting an Interaction Interface

Jastrzębska

Chen

Orban

et al. 2015

Journal of Biological Chemistry

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Although homo- and heterodimerizations of G protein-coupled receptors (GPCRs) are well documented, GPCR monomers are able to assemble in different ways, thus causing variations in the interactive interface between receptor monomers among different GPCRs. Moreover, the functional consequences of this phenomenon, which remain to be clarified, could be specific for different GPCRs. Synthetic peptides derived from transmembrane (TM) domains can interact with a full-length GPCR, blocking dimer formation and affecting its function. Here we used peptides corresponding to TM helices of bovine rhodopsin (Rho) to investigate the Rho dimer interface and functional consequences of its disruption. Incubation of Rho with TM1, TM2, TM4, and TM5 peptides in rod outer segment (ROS) membranes shifted the resulting detergent-solubilized protein migration through a gel filtration column toward smaller molecular masses with a reduced propensity for dimer formation in a cross-linking reaction. Binding of these TM peptides to Rho was characterized by both mass spectrometry and a label-free assay from which dissociation constants were calculated. A BRET (bioluminescence resonance energy transfer) assay revealed that the physical interaction between Rho molecules expressed in membranes of living cells was blocked by the same four TM peptides identified in our in vitro experiments. Although disruption of the Rho dimer/oligomer had no effect on the rates of G protein activation, binding of Gt to the activated receptor stabilized the dimer. However, TM peptide-induced disruption of dimer/oligomer decreased receptor stability, suggesting that Rho supramolecular organization could be essential for ROS stabilization and receptor trafficking.

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Analysis of Human Dopamine D3 Receptor Quaternary Structure

Cited by 23 publications

References 54 publications

The dopamine D2 receptor dimer and its interaction with homobivalent antagonists: homology modeling, docking and molecular dynamics

The dopamine D2 receptor dimer and its interaction with homobivalent antagonists: homology modeling, docking and molecular dynamics

A G Protein-Coupled Receptor Dimerization Interface in Human Cone Opsins

Disruption of Rhodopsin Dimerization with Synthetic Peptides Targeting an Interaction Interface

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