Dualsteric Muscarinic Antagonists–Orthosteric Binding Pose Controls Allosteric Subtype Selectivity

Schmitz, Jens; Mey, Dorina van der; Bermudez, Marcel; Klöckner, Jessica; Schrage, R; Kostenis, Evi; Tränkle, Christian; Wolber, Gerhard; Mohr, Klaus; Holzgrabe, Ulrike

doi:10.1021/jm500790x

Cited by 33 publications

(44 citation statements)

References 58 publications

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“…Modes of a Dualsteric Agonist-Dualsteric (also termed bitopic orthosteric/allosteric) ligands (30) for muscarinic receptors are composed of orthosteric and allosteric moieties covalently connected via linkers of different chemical nature (31)(32)(33)(34). Recent studies suggested that at least some of these bipharmacophoric ligands may have more than one binding mode (28,34).…”

Section: Direct Pharmacological Evidence For Multiple Bindingmentioning

confidence: 99%

“…Unlike previous applications of gathering pharmacophore information from molecular dynamics (33,69), this new implementation works in a fully automated way: dynophore groups interaction points (such as hydrogen bonds, charges, and lipophilic contacts) of each trajectory frame according to their type and ligand atoms involved. All feature groups are graphically represented by three-dimensional volumetric feature density clouds, which are statistically characterized by occurrence frequency and interaction patterns with the protein.…”

Section: Site-directed Mutagenesis and Generation Of Stable Cellmentioning

confidence: 99%

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Ligand Binding Ensembles Determine Graded Agonist Efficacies at a G Protein-coupled Receptor

Böck

Bermudez

Krebs

et al. 2016

Journal of Biological Chemistry

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G protein-coupled receptors constitute the largest family of membrane receptors and modulate almost every physiological process in humans. Binding of agonists to G protein-coupled receptors induces a shift from inactive to active receptor conformations. Biophysical studies of the dynamic equilibrium of receptors suggest that a portion of receptors can remain in inactive states even in the presence of saturating concentrations of agonist and G protein mimetic. However, the molecular details of agonist-bound inactive receptors are poorly understood.Here we use the model of bitopic orthosteric/allosteric (i.e. dualsteric) agonists for muscarinic M 2 receptors to demonstrate the existence and function of such inactive agonist⅐receptor complexes on a molecular level. Using all-atom molecular dynamics simulations, dynophores (i.e. a combination of static three-dimensional pharmacophores and molecular dynamics-based conformational sampling), ligand design, and receptor mutagenesis, we show that inactive agonist⅐receptor complexes can result from agonist binding to the allosteric vestibule alone, whereas the dualsteric binding mode produces active receptors. Each agonist forms a distinct ligand binding ensemble, and different agonist efficacies depend on the fraction of purely allosteric (i.e. inactive) versus dualsteric (i.e. active) binding modes. We propose that this concept may explain why agonist⅐receptor complexes can be inactive and that adopting multiple binding modes may be generalized also to small agonists where binding modes will be only subtly different and confined to only one binding site.Specific and coordinated cell-to-cell communication regulates the flow of information between cells, and proper information processing ensures physiological functions of biological systems. G protein-coupled receptors (GPCRs), 8 constituting the largest class of membrane proteins in mammals, are essential mediators of chemically and light-encoded information (1-4). GPCRs sense a great variety of extracellular stimuli, e.g. neurotransmitters and hormones, and subsequently translate this information into an intracellular response via G proteins, ␤-arrestins, and possibly GPCR-interacting proteins (2-5). Because of their abundance and relevance in regulating the majority of (patho-)physiological processes in humans, GPCRs have for a long time represented the most important drug targets being addressed by at least a third of all currently marketed drugs (6, 7).Agonist binding leads to receptor activation, which is followed by intracellular G protein recruitment and subsequent cell signaling. Breakthroughs in GPCR crystallography have led to inactive and active crystal structures of the same receptor protein. Among these are rhodopsin (8 -10) and more recently the ␤ 2 -adrenergic (11-14), M 2 muscarinic (15, 16), and -opioid receptors (17, 18). These structures most likely represent energetically favored, relatively stable inactive and active receptor⅐ligand complexes. Despite the diversity of crystallized receptors, a common...

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Section: Direct Pharmacological Evidence For Multiple Bindingmentioning

confidence: 99%

Section: Site-directed Mutagenesis and Generation Of Stable Cellmentioning

confidence: 99%

Ligand Binding Ensembles Determine Graded Agonist Efficacies at a G Protein-coupled Receptor

Böck

Bermudez

Krebs

et al. 2016

Journal of Biological Chemistry

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“…Due to available crystal structures and mutational data, the M 2 receptor is a classical example in many studies 6e,11. The identified key residues for allosteric binding are Y177 and W422.…”

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confidence: 99%

Structural Characteristics of the Allosteric Binding Site Represent a Key to Subtype Selective Modulators of Muscarinic Acetylcholine Receptors

Bermudez

Rakers

Wolber

2015

Molecular Informatics

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The high conservation of the orthosteric acetylcholine binding site of muscarinic receptors (MAChR) represents a considerable challenge in terms of designing subtype selective drugs. A promising approach to gain subtype selectivity is to include allosteric or dualsteric targeting that aims to address more specific extracellular binding sites. Despite recent advances in crystallography of G protein coupled receptors (GPCRs), structural information for all 5 MAChR subtypes is not yet available. Here we report structural models of the active and the inactive receptor state of all subtypes derived by homology modelling in combination with MD simulations. The comparison of the allosteric binding site unveils the characteristics for each subtype on a structural level and indicates anchor points for rational design of selective drugs. Additionally, homology models offer the possibility for a rational explanation of dualsteric subtype selectivity, as we show for the M2 over M5 selectivity of the dualsteric ligands Atr-6-naph and Iper-6-phth.

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“…Bitopic ligands, also described as ‘dualsteric’, bridge two different sites on a single GPCR, such as an orthosteric agonist site and an allosteric enhancer site [98,128]. A biotopic antagonist of the M 2 muscarinic receptor demonstrated that the orthosteric binding portion directs the allosteric moiety to establish its subtype selectivity [147]. …”

Section: New Pharmacological Dimensionsmentioning

confidence: 99%

New paradigms in GPCR drug discovery

Jacobson

2015

Biochemical Pharmacology

150

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G protein-coupled receptors (GPCRs) remain a major domain of pharmaceutical discovery. The discovery of GPCR lead compounds and their optimization are now structure-based, thanks to advances in X-ray crystallography, molecular modeling, protein engineering and biophysical techniques. In silico screening provides useful hit molecules. New pharmacological approaches to tuning the pleotropic action of GPCRs include: allosteric modulators, biased ligands, GPCR heterodimer-targeted compounds, manipulation of polypharmacology, receptor antibodies and tailoring of drug molecules to fit GPCR pharmacogenomics. Measurements of kinetics and drug efficacy are factors influencing clinical success. With the exception of inhibitors of GPCR kinases, targeting of intracellular GPCR signaling or receptor cycling for therapeutic purposes remains a futuristic concept. New assay approaches are more efficient and multidimensional: cell-based, label-free, fluorescence-based assays, and biosensors. Tailoring GPCR drugs to a patient’s genetic background is now being considered. Chemoinformatic tools can predict ADME-tox properties. New imaging technology visualizes drug action in vivo. Thus, there is reason to be optimistic that new technology for GPCR ligand discovery will help improve the current narrowing of the pharmaceutical pipeline.

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Dualsteric Muscarinic Antagonists–Orthosteric Binding Pose Controls Allosteric Subtype Selectivity

Cited by 33 publications

References 58 publications

Ligand Binding Ensembles Determine Graded Agonist Efficacies at a G Protein-coupled Receptor

Ligand Binding Ensembles Determine Graded Agonist Efficacies at a G Protein-coupled Receptor

Structural Characteristics of the Allosteric Binding Site Represent a Key to Subtype Selective Modulators of Muscarinic Acetylcholine Receptors

New paradigms in GPCR drug discovery

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