MS Binding Assays for Glycine Transporter 2 (GlyT2) Employing Org25543 As Reporter Ligand

Overview

Journal ChemMedChem

Specialties Chemistry
Pharmacology

Date 2020 Aug 1

PMID 32734692

Citations 2

Authors

Thomas M Ackermann

Lars Allmendinger

Georg Hofner

Klaus T Wanner

Affiliations

Soon will be listed here.

Abstract

This study describes the first binding assay for glycine transporter 2 (GlyT2) following the concept of MS Binding Assays. The selective GlyT2 inhibitor Org25543 was employed as a reporter ligand and it was quantified with a highly sensitive and rapid LC-ESI-MS/MS method. Binding of Org25543 at GlyT2 was characterized in kinetic and saturation experiments with an off-rate of 7.07×10 s , an on-rate of 1.01×10 M s , and an equilibrium dissociation constant of 7.45 nM. Furthermore, the inhibitory constants of 19 GlyT ligands were determined in competition experiments. The validity of the GlyT2 affinities determined with the binding assay was examined by a comparison with published inhibitory potencies from various functional assays. With the capability for affinity determination towards GlyT2 the developed MS Binding Assays provide the first tool for affinity profiling of potential ligands and it represents a valuable new alternative to functional assays addressing GlyT2.

Citing Articles

Screening for New Inhibitors of Glycine Transporter 1 and 2 by Means of MS Binding Assays.

Ackermann T, Hofner G, Wanner K ChemMedChem. 2021; 16(19):3094-3104.

PMID: 34174033 PMC: 8518836. DOI: 10.1002/cmdc.202100408.

MS Binding Assays for Glycine Transporter 2 (GlyT2) Employing Org25543 as Reporter Ligand.

Ackermann T, Allmendinger L, Hofner G, Wanner K ChemMedChem. 2020; 16(1):199-215.

PMID: 32734692 PMC: 7821181. DOI: 10.1002/cmdc.202000342.

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