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

Jack, Thomas; Leuenberger, Michele; Ruepp, Marc-David; Vernekar, Sanjeev Kumar V; Thompson, Andrew James; Braga-Lagache, Sophie; Heller, Manfred; Lochner, Martin (2019). Mapping the Orthosteric Binding Site of the Human 5-HT3 Receptor Using Photo-crosslinking Antagonists. ACS chemical neuroscience, 10(1), pp. 438-450. American Chemical Society 10.1021/acschemneuro.8b00327

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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.

Item Type:

Journal Article (Original Article)

Division/Institute:

04 Faculty of Medicine > Pre-clinic Human Medicine > BioMedical Research (DBMR) > DCR Services > Core Facility Massenspektrometrie- und Proteomics-Labor
04 Faculty of Medicine > Pre-clinic Human Medicine > BioMedical Research (DBMR) > Unit Childrens Hospital > Protein- und Zellbiologie
04 Faculty of Medicine > Pre-clinic Human Medicine > BioMedical Research (DBMR)
08 Faculty of Science > Departement of Chemistry and Biochemistry
04 Faculty of Medicine > Pre-clinic Human Medicine > Institute of Biochemistry and Molecular Medicine

UniBE Contributor:

Jack, Thomas; Leuenberger, Michele; Ruepp, Marc-David; Thompson, Andrew James; Braga, Sophie Marie-Pierre; Heller, Manfred and Lochner, Martin

Subjects:

600 Technology > 610 Medicine & health
500 Science > 570 Life sciences; biology
500 Science > 540 Chemistry

ISSN:

1948-7193

Publisher:

American Chemical Society

Funders:

[4] Swiss National Science Foundation
[100] Holcim
[91] British Heart Foundation

Language:

English

Submitter:

Martin Lochner

Date Deposited:

29 Aug 2018 10:31

Last Modified:

01 Sep 2019 02:32

Publisher DOI:

10.1021/acschemneuro.8b00327

PubMed ID:

30149702

BORIS DOI:

10.7892/boris.119622

URI:

https://boris.unibe.ch/id/eprint/119622

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