Mapping the Orthosteric Binding Site of the Human 5-HT<sub>3</sub> Receptor Using Photo-cross-linking Antagonists

Jack, Thomas R.; Leuenberger, Michele; Ruepp, Marc‐David; Vernekar, Sanjeev Kumar V.; Thompson, Andrew; Braga-Lagache, Sophie; Heller, Manfred; Lochner, Martin

doi:10.1021/acschemneuro.8b00327

Cited by 7 publications

(9 citation statements)

References 47 publications

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“…The structural studies have, to a large degree, refined and elaborated on the molecular understanding of the orthosteric sites initially characterized through mutagenesis and ligand binding approaches [ 15 , 19 , 150 , 151 ]. The three-dimensional structures open up many opportunities to employ additional techniques such as molecular dynamics simulations [ 141 , 142 , 145 , 146 ] in conjunction with photo-crosslinking and mass spectrometry to delve further into the dynamics of ligand receptor interactions [ 152 ]. The transmembrane domains are arranged asymmetrically so transmembrane domain 2 (TM2) faces the pore and contributes most to ion interactions therein ( Figure 2 ).…”

Section: Structural Insights Into 5-ht 3 Receptor Actionmentioning

confidence: 99%

Tapping into 5-HT3 Receptors to Modify Metabolic and Immune Responses

Irving

Turek

Kettle

et al. 2021

IJMS

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5-hydroxytryptamine type 3 (5-HT3) receptors are ligand gated ion channels, which clearly distinguish their mode of action from the other G-protein coupled 5-HT or serotonin receptors. 5-HT3 receptors are well established targets for emesis and gastrointestinal mobility and are used as adjunct targets in treating schizophrenia. However, the distribution of these receptors is wider than the nervous system and there is potential that these additional sites can be targeted to modulate inflammatory and/or metabolic conditions. Recent progress in structural biology and pharmacology of 5-HT3 receptors have provided profound insights into mechanisms of their action. These advances, combined with insights into clinical relevance of mutations in genes encoding 5-HT3 subunits and increasing understanding of their implications in patient’s predisposition to diseases and response to the treatment, open new avenues for personalized precision medicine. In this review, we recap on the current status of 5-HT3 receptor-based therapies using a biochemical and physiological perspective. We assess the potential for targeting 5-HT3 receptors in conditions involving metabolic or inflammatory disorders based on recent findings, underscoring the challenges and limitations of this approach.

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Section: Structural Insights Into 5-ht 3 Receptor Actionmentioning

confidence: 99%

Tapping into 5-HT3 Receptors to Modify Metabolic and Immune Responses

Irving

Turek

Kettle

et al. 2021

IJMS

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“…The human C and E subunits also influence the electrical properties in response to 5-HT 3 receptor agonists and antagonists [ 81 , 82 ]. In contrast to humans, rodents only express A and B subunits just forming 5HT 3 A receptor homomers or 5-HT 3 AB receptor heteromers [ 80 , 85 ].…”

Section: Introductionmentioning

confidence: 99%

“…We identified that all subunits are present in some members of Chordata, Mollusca, Platyhelminthes, and Cnidaria. As the A subunit is particularly important in forming the orthosteric binding site [ 85 , 86 ], we investigated how ligand binding components (amino acid residues) in this subunit are altered across the phyla.…”

Section: Introductionmentioning

confidence: 99%

Phylogenetic analyses of 5-hydroxytryptamine 3 (5-HT3) receptors in Metazoa

Rao

Turek²,

Irving³

2023

PLoS ONE

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The 5-hydroxytrptamine 3 (5-HT3) receptor is a member of the ’Cys-loop’ family and the only pentameric ligand gated ion channel among the serotonin receptors. 5-HT3 receptors play an important role in controlling growth, development, and behaviour in animals. Several 5-HT3 receptor antagonists are used to treat diseases (e.g., irritable bowel syndrome, nausea and emesis). Humans express five different subunits (A-E) enabling a variety of heteromeric receptors to form but all contain 5HT3A subunits. However, the information available about the 5-HT3 receptor subunit occurrence among the metazoan lineages is minimal. In the present article we searched for 5-HT3 receptor subunit homologs from different phyla in Metazoa. We identified more than 1000 5-HT3 receptor subunits in Metazoa in different phyla and undertook simultaneous phylogenetic analysis of 526 5HT3A, 358 5HT3B, 239 5HT3C, 70 5HT3D, and 173 5HT3E sequences. 5-HT3 receptor subunits were present in species belonging to 11 phyla: Annelida, Arthropoda, Chordata, Cnidaria, Echinodermata, Mollusca, Nematoda, Orthonectida, Platyhelminthes, Rotifera and Tardigrada. All subunits were most often identified in Chordata phylum which was strongly represented in searches. Using multiple sequence alignment, we investigated variations in the ligand binding region of the 5HT3A subunit protein sequences in the metazoan lineage. Several critical amino acid residues important for ligand binding (common structural features) are commonly present in species from Nematoda and Platyhelminth gut parasites through to Chordata. Collectively, this better understanding of the 5-HT3 receptor evolutionary patterns raises possibilities of future pharmacological challenges facing Metazoa including effects on parasitic and other species in ecosystems that contain 5-HT3 receptor ligands.

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“…These works are enabled by the strategies used in activity-based protein profiling (ABPP), photoaffinity labeling, and advances in mass spectrometry . This strategy has recently been used to great effect to study the activity profiles of NSAIDs, cannabinoid drugs, and methamphetamine, for example. − Detailed receptor binding site studies have been enabled by photo-cross-linking the CNS drugs granisetron, propofol, glutamate receptor modulators, and others. − DRD2 itself has a long history of use with photoaffinity technology to aid the biochemical characterization of this hard-to-handle membrane-bound protein. , …”

Section: Introductionmentioning

confidence: 99%

Developing Photoaffinity Probes for Dopamine Receptor D₂ to Determine Targets of Parkinson’s Disease Drugs

Kim

Doukmak

Flax

et al. 2022

ACS Chem. Neurosci.

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Dopaminergic pathways control highly consequential aspects of physiology and behavior. One of the most therapeutically important and best-studied receptors in these pathways is dopamine receptor D2 (DRD2). Unfortunately, DRD2 is challenging to study with traditional molecular biological techniques, and most drugs designed to target DRD2 are ligands for many other receptors. Here, we developed probes able to both covalently bind to DRD2 using photoaffinity labeling and provide a chemical handle for detection or affinity purification. These probes behaved like good DRD2 agonists in traditional biochemical assays and were able to perform in chemical–biological assays of cell and receptor labeling. Rat whole brain labeling and affinity enrichment using the probes permitted proteomic analysis of the probes’ interacting proteins. Bioinformatic study of the hits revealed that the probes bound noncanonically targeted proteins in Parkinson’s disease network as well as the retrograde endocannabinoid signaling, neuronal nitric oxide synthase, muscarinic acetylcholine receptor M1, GABA receptor, and dopamine receptor D1 (DRD1) signaling networks. Follow-up analysis may yield insights into how this pathway relates specifically to Parkinson’s disease symptoms or provide new targets for treatments. This work reinforces the notion that the combination of chemical biology and omics-based approaches provides a broad picture of a molecule’s “interactome” and may also give insight into the pleiotropy of effects observed for a drug or perhaps indicate new applications.

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Mapping the Orthosteric Binding Site of the Human 5-HT₃ Receptor Using Photo-cross-linking Antagonists

Cited by 7 publications

References 47 publications

Tapping into 5-HT3 Receptors to Modify Metabolic and Immune Responses

Tapping into 5-HT3 Receptors to Modify Metabolic and Immune Responses

Phylogenetic analyses of 5-hydroxytryptamine 3 (5-HT3) receptors in Metazoa

Developing Photoaffinity Probes for Dopamine Receptor D₂ to Determine Targets of Parkinson’s Disease Drugs

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Mapping the Orthosteric Binding Site of the Human 5-HT3 Receptor Using Photo-cross-linking Antagonists

Cited by 7 publications

References 47 publications

Tapping into 5-HT3 Receptors to Modify Metabolic and Immune Responses

Tapping into 5-HT3 Receptors to Modify Metabolic and Immune Responses

Phylogenetic analyses of 5-hydroxytryptamine 3 (5-HT3) receptors in Metazoa

Developing Photoaffinity Probes for Dopamine Receptor D2 to Determine Targets of Parkinson’s Disease Drugs

Contact Info

Product

Resources

About

Mapping the Orthosteric Binding Site of the Human 5-HT₃ Receptor Using Photo-cross-linking Antagonists

Developing Photoaffinity Probes for Dopamine Receptor D₂ to Determine Targets of Parkinson’s Disease Drugs