Mapping the Orthosteric Binding Site of the Human 5-HT Receptor Using Photo-crosslinking Antagonists

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

doi:10.1021/acschemneuro.8b00327

Mapping the Orthosteric Binding Site of the Human 5-HT Receptor Using Photo-crosslinking Antagonists

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

Basic and Clinical Neuroscience

Research output: Contribution to journal › Article › peer-review

5 Citations (Scopus)

Abstract

The serotonin-gated 5-HT3 receptor is a ligand-gated ion channel. Its location at the synapse in the central and peripheral nervous system have rendered it a prime pharmacological target, e.g. for antiemetic drugs that bind with high affinity to the neurotransmitter binding site and prevent the opening of the channel. Advances in structural biology techniques has led to a surge of disclosed three-dimensional receptor structures, however, solving ligand-bound high-resolution 5-HT3 receptor structures has not been achieved to date. Ligand binding poses in the orthosteric binding site have been largely predicted from mutagenesis and docking studies. We report the synthesis of a series of photo-crosslinking compounds whose structures are based on the clinically used antiemetic drug granisetron (Kytril®). These displaced [3H]granisetron from the orthosteric binding site with low nanomolar affinities and showed specific photo-crosslinking with the human 5-HT3 receptor. Detailed analysis by protein-MS/MS identified a residue (Met-228) near the tip of binding loop C as the covalent modification site.

Original language	English
Journal	Acs chemical neuroscience
DOIs	https://doi.org/10.1021/acschemneuro.8b00327
Publication status	E-pub ahead of print - 27 Aug 2018

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title = "Mapping the Orthosteric Binding Site of the Human 5-HT Receptor Using Photo-crosslinking Antagonists",

abstract = "The serotonin-gated 5-HT3 receptor is a ligand-gated ion channel. Its location at the synapse in the central and peripheral nervous system have rendered it a prime pharmacological target, e.g. for antiemetic drugs that bind with high affinity to the neurotransmitter binding site and prevent the opening of the channel. Advances in structural biology techniques has led to a surge of disclosed three-dimensional receptor structures, however, solving ligand-bound high-resolution 5-HT3 receptor structures has not been achieved to date. Ligand binding poses in the orthosteric binding site have been largely predicted from mutagenesis and docking studies. We report the synthesis of a series of photo-crosslinking compounds whose structures are based on the clinically used antiemetic drug granisetron (Kytril{\textregistered}). These displaced [3H]granisetron from the orthosteric binding site with low nanomolar affinities and showed specific photo-crosslinking with the human 5-HT3 receptor. Detailed analysis by protein-MS/MS identified a residue (Met-228) near the tip of binding loop C as the covalent modification site.",

author = "Thomas Jack and Michele Leuenberger and Marc-David Ruepp and Vernekar, {Sanjeev Kumar V} and Thompson, {Andrew J} and Sophie Braga-Lagache and Manfred Heller and Martin Lochner",

year = "2018",

month = aug,

day = "27",

doi = "10.1021/acschemneuro.8b00327",

language = "English",

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AU - Jack, Thomas

AU - Leuenberger, Michele

AU - Ruepp, Marc-David

AU - Vernekar, Sanjeev Kumar V

AU - Thompson, Andrew J

AU - Braga-Lagache, Sophie

AU - Heller, Manfred

AU - Lochner, Martin

PY - 2018/8/27

Y1 - 2018/8/27

N2 - The serotonin-gated 5-HT3 receptor is a ligand-gated ion channel. Its location at the synapse in the central and peripheral nervous system have rendered it a prime pharmacological target, e.g. for antiemetic drugs that bind with high affinity to the neurotransmitter binding site and prevent the opening of the channel. Advances in structural biology techniques has led to a surge of disclosed three-dimensional receptor structures, however, solving ligand-bound high-resolution 5-HT3 receptor structures has not been achieved to date. Ligand binding poses in the orthosteric binding site have been largely predicted from mutagenesis and docking studies. We report the synthesis of a series of photo-crosslinking compounds whose structures are based on the clinically used antiemetic drug granisetron (Kytril®). These displaced [3H]granisetron from the orthosteric binding site with low nanomolar affinities and showed specific photo-crosslinking with the human 5-HT3 receptor. Detailed analysis by protein-MS/MS identified a residue (Met-228) near the tip of binding loop C as the covalent modification site.

AB - The serotonin-gated 5-HT3 receptor is a ligand-gated ion channel. Its location at the synapse in the central and peripheral nervous system have rendered it a prime pharmacological target, e.g. for antiemetic drugs that bind with high affinity to the neurotransmitter binding site and prevent the opening of the channel. Advances in structural biology techniques has led to a surge of disclosed three-dimensional receptor structures, however, solving ligand-bound high-resolution 5-HT3 receptor structures has not been achieved to date. Ligand binding poses in the orthosteric binding site have been largely predicted from mutagenesis and docking studies. We report the synthesis of a series of photo-crosslinking compounds whose structures are based on the clinically used antiemetic drug granisetron (Kytril®). These displaced [3H]granisetron from the orthosteric binding site with low nanomolar affinities and showed specific photo-crosslinking with the human 5-HT3 receptor. Detailed analysis by protein-MS/MS identified a residue (Met-228) near the tip of binding loop C as the covalent modification site.

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DO - 10.1021/acschemneuro.8b00327

M3 - Article

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SN - 1948-7193

JO - Acs chemical neuroscience

JF - Acs chemical neuroscience

ER -

Mapping the Orthosteric Binding Site of the Human 5-HT Receptor Using Photo-crosslinking Antagonists

Abstract

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