9HUF

Outward-open structure of human glycine transporter 2 bound to allosteric inhibitor RPI-GLYT2-82

This is a non-PDB format compatible entry.

Summary for 9HUF

• Entry DOI: 10.2210/pdb9huf/pdb
• EMDB information: 52409 52410 52411
• Descriptor: Sodium- and chloride-dependent glycine transporter 2, CHLORIDE ION, ~{N}-[[4-(dimethylamino)oxan-4-yl]methyl]-1-(phenylcarbonyl)indole-5-carboxamide, ... (7 entities in total)
• Functional Keywords: transport protein, slc6a5, neurotransmitter/sodium symporter, sodium- and chloride-dependent glycine transporter 2, membrane protein
• Biological source: Homo sapiens (human)
• Total number of polymer chains: 1
• Total formula weight: 70699.99

Authors

Cantwell Chater, R.P.,Peiser-Oliver, J.,Pati, T.K.,Quinn, A.S.,Lotsaris, I.,Frangos, Z.J.,Anderson, K.E.,Tischer, A.E.,Williams-Noonan, B.J.,Aubrey, K.R.,O Mara, M.L.,Michaelides, M.,Mohammadi, S.A.,Cioffi, C.L.,Vandenberg, R.J.,Shahsavar, A. (deposition date: 2024-12-22, release date: 2026-02-25, Last modification date: 2026-03-25)

Primary citation

Cantwell Chater, R.P.,Peiser-Oliver, J.,Pati, T.K.,Quinn, A.S.,Lotsaris, I.,Frangos, Z.J.,Anderson, K.E.,Tischer, A.E.,Williams-Noonan, B.J.,Aubrey, K.R.,O'Mara, M.L.,Michaelides, M.,Mohammadi, S.A.,Cioffi, C.L.,Vandenberg, R.J.,Shahsavar, A.
A reversible allosteric inhibitor of GlyT2 alleviates neuropathic pain without on-target side effects.
Biorxiv, 2025

PubMed Abstract: Chronic neuropathic pain, caused by nerve damage or disease, is increasing in prevalence, but current treatments are ineffective and over-reliant on opioids. The neuronal glycine transporter, GlyT2, regulates inhibitory glycinergic neurotransmission and represents a promising target for new analgesics. However, most GlyT2 inhibitors cause significant side effects, in part due to irreversible inhibition at analgesic doses. Here we develop a reversible inhibitor of GlyT2, RPI-GLYT2-82, and identify its binding site by determining cryo-EM structures of human GlyT2. We capture three fundamental conformational states of GlyT2 in the substrate-free state, and bound to either glycine, RPI-GLYT2-82 or the pseudo-irreversible inhibitor ORG25543. We demonstrate that RPI-GLYT2-82 dissociates from GlyT2 faster than ORG25543, providing analgesia in mouse neuropathic pain models without on-target side-effects or addiction liability. Our data provide a mechanistic understanding of allosteric inhibition of glycine transport, enabling structure-based design of non-opioid analgesics.

PubMed: 41332641
DOI: 10.1101/2025.04.21.649698

Experimental method

ELECTRON MICROSCOPY (2.79 Å)