Harvey, R. J. & Yee, B. K. Glycine transporters as novel therapeutic targets in schizophrenia, alcohol dependence and pain. Nat. Rev. Drug Discov. 12, 866–885 (2013).
Google Scholar
Grenningloh, G. et al. The strychnine-binding subunit of the glycine receptor shows homology with nicotinic acetylcholine receptors. Nature 328, 215–220 (1987).
Google Scholar
Johnson, J. W. & Ascher, P. Glycine potentiates the NMDA response in cultured mouse brain neurons. Nature 325, 529–531 (1987).
Google Scholar
Cioffi, C. L. Glycine transporter-1 inhibitors: a patent review (2011–2016). Expert Opin. Ther. Pat. 28, 197–210 (2018).
Google Scholar
Kristensen, A. S. et al. SLC6 neurotransmitter transporters: structure, function, and regulation. Pharmacol. Rev. 63, 585–640 (2011).
Google Scholar
Gomeza, J. et al. Inactivation of the glycine transporter 1 gene discloses vital role of glial glycine uptake in glycinergic inhibition. Neuron 40, 785–796 (2003).
Google Scholar
Cubelos, B., Giménez, C. & Zafra, F. Localization of the GLYT1 glycine transporter at glutamatergic synapses in the rat brain. Cereb. Cortex 15, 448–459 (2005).
Google Scholar
Cubelos, B., González-González, I. M., Giménez, C. & Zafra, F. The scaffolding protein PSD-95 interacts with the glycine transporter GLYT1 and impairs its internalization. J. Neurochem. 95, 1047–1058 (2005).
Google Scholar
Kantrowitz, J. T. & Javitt, D. C. N-methyl-d-aspartate (NMDA) receptor dysfunction or dysregulation: the final common pathway on the road to schizophrenia? Brain Res. Bull. 83, 108–121 (2010).
Google Scholar
Pinard, E., Borroni, E., Koerner, A., Umbricht, D. & Alberati, D. Glycine transporter type I (GlyT1) inhibitor, bitopertin: a journey from lab to patient. CHIMIA Int. J. Chem. 72, 477–484 (2018).
Google Scholar
Shim, S. S., Hammonds, M. D. & Kee, B. S. Potentiation of the NMDA receptor in the treatment of schizophrenia: focused on the glycine site. Eur. Arch. Psychiatry Clin. Neurosci. 258, 16–27 (2007).
Google Scholar
Pinard, E. et al. Selective GlyT1 inhibitors: discovery of [4-(3-fluoro-5-trifluoromethylpyridin-2-yl)piperazin-1-yl][5-methanesulfonyl-2-((S)-2,2,2-trifluoro-1-methylethoxy)phenyl]methanone (RG1678), a promising novel medicine to treat schizophrenia. J. Med. Chem. 53, 4603–4614 (2010).
Google Scholar
Krystal, J. H. et al. Neuroplasticity as a target for the pharmacotherapy of anxiety disorders, mood disorders, and schizophrenia. Drug Discov. Today 14, 690–697 (2009).
Google Scholar
D’Souza, D. C. et al. Dose-related target occupancy and effects on circuitry, behavior, and neuroplasticity of the glycine transporter-1 inhibitor PF-03463275 in healthy and schizophrenia subjects. Biol. Psychiatry 84, 413–421 (2018).
Google Scholar
Jardetzky, O. Simple allosteric model for membrane pumps. Nature 211, 969–970 (1966).
Google Scholar
Kazmier, K. et al. Conformational dynamics of ligand-dependent alternating access in LeuT. Nat. Struct. Mol. Biol. 21, 472–479 (2014).
Google Scholar
Malinauskaite, L. et al. A mechanism for intracellular release of Na+ by neurotransmitter/sodium symporters. Nat. Struct. Mol. Biol. 21, 1006–1012 (2014).
Google Scholar
Penmatsa, A., Wang, K. H. & Gouaux, E. X-ray structure of dopamine transporter elucidates antidepressant mechanism. Nature 503, 85–90 (2013).
Google Scholar
Coleman, J. A. et al. Serotonin transporter–ibogaine complexes illuminate mechanisms of inhibition and transport. Nature 569, 141–145 (2019).
Google Scholar
Gotfryd, K. et al. X-ray structure of LeuT in an inward-facing occluded conformation reveals mechanism of substrate release. Nat. Commun. 11, 1005 (2020).
Google Scholar
Singh, S. K., Yamashita, A. & Gouaux, E. Antidepressant binding site in a bacterial homologue of neurotransmitter transporters. Nature 448, 952–956 (2007).
Google Scholar
Coleman, J. A., Green, E. M. & Gouaux, E. X-ray structures and mechanism of the human serotonin transporter. Nature 532, 334–339 (2016).
Google Scholar
Malinauskaite, L. et al. A conserved leucine occupies the empty substrate site of LeuT in the Na+-free return state. Nat. Commun. 7, 11673 (2016).
Google Scholar
Alberati, D. et al. Glycine reuptake inhibitor RG1678: a pharmacologic characterization of an investigational agent for the treatment of schizophrenia. Neuropharmacology 62, 1152–1161 (2012).
Google Scholar
Pinard, E. et al. Discovery of benzoylisoindolines as a novel class of potent, selective and orally active GlyT1 inhibitors. Bioorg. Med. Chem. Lett. 20, 6960–6965 (2010).
Google Scholar
Jolidon, S., Narquizian, R., Norcross, R. D. & Pinard, E. Heterocyclic substituted phenyl methanones as inhibitors of the glycine transporter 1. WIPO patent WO/2006/082001 (2006).
Brown, A. et al. Discovery and SAR of Org 24598—a selective glycine uptake inhibitor. Bioorg. Med. Chem. Lett. 11, 2007–2009 (2001).
Google Scholar
Zimmermann, I. et al. Synthetic single domain antibodies for the conformational trapping of membrane proteins. eLife 7, e34317 (2018).
Google Scholar
Abramson, J. & Wright, E. M. Structure and function of Na+-symporters with inverted repeats. Curr. Opin. Struct. Biol. 19, 425–432 (2009).
Google Scholar
LeVine, M. V. et al. The allosteric mechanism of substrate-specific transport in SLC6 is mediated by a volumetric sensor. Proc. Natl Acad. Sci. USA 116, 15947–15956 (2019).
Google Scholar
Carland, J. E. et al. Molecular determinants for substrate interactions with the glycine transporter GlyT2. ACS Chem. Neurosci. 9, 603–614 (2018).
Google Scholar
Focht, D. et al. A non-helical region in transmembrane helix 6 of hydrophobic amino acid transporter MhsT mediates substrate recognition. EMBO J. 40, e105164 (2020).
Google Scholar
Jaeger, K. et al. Structural basis for allosteric ligand recognition in the human CC chemokine receptor 7. Cell 178, 1222–1230 (2019).
Google Scholar
Vandenberg, R. J., Shaddick, K. & Ju, P. Molecular basis for substrate discrimination by glycine transporters. J. Biol. Chem. 282, 14447–14453 (2007).
Google Scholar
Werdehausen, R. et al. Lidocaine metabolites inhibit glycine transporter 1: a novel mechanism for the analgesic action of systemic lidocaine? Anesthesiology 116, 147–158 (2012).
Google Scholar
Jacobs, M. T., Zhang, Y.-W., Campbell, S. D. & Rudnick, G. Ibogaine, a noncompetitive inhibitor of serotonin transport, acts by stabilizing the cytoplasm-facing state of the transporter. J. Biol. Chem. 282, 29441–29447 (2007).
Google Scholar
Bugarski-Kirola, D. et al. Bitopertin in negative symptoms of schizophrenia-results from the phase III FlashLyte and DayLyte studies. Biol. Psychiatry 82, 8–16 (2017).
Google Scholar
Martin-Facklam, M. et al. Glycine transporter type 1 occupancy by bitopertin: a positron emission tomography study in healthy volunteers. Neuropsychopharmacology 38, 504–512 (2013).
Google Scholar
Weber, F. et al. Brain shuttle antibody for Alzheimer’s disease with attenuated peripheral effector function due to an inverted binding mode. Cell Rep. 22, 149–162 (2018).
Google Scholar
Olivares, L., Aragón, C., Giménez, C. & Zafra, F. The role of N-glycosylation in the targeting and activity of the GLYT1 glycine transporter. J. Biol. Chem. 270, 9437–9442 (1995).
Google Scholar
Gati, C. et al. Serial crystallography on in vivo grown microcrystals using synchrotron radiation. IUCrJ. 1, 87–94 (2014).
Google Scholar
Zander, U. et al. MeshAndCollect: an automated multi-crystal data-collection workflow for synchrotron macromolecular crystallography beamlines. Acta Crystallogr. D 71, 2328–2343 (2015).
Google Scholar
Popov, A. N. & Bourenkov, G. Dozor (European Synchrotron Radiation Facility, 2016).
Tange, O. GNU Parallel: the command-line power tool. The USENIX Magazine 36, 42–47 (2011).
Kabsch, W. XDS. Acta Crystallogr. D 66, 125–132 (2010).
Google Scholar
Emsley, P., Lohkamp, B., Scott, W. G. & Cowtan, K. Features and development of Coot. Acta Crystallogr. D 66, 486–501 (2010).
Google Scholar
Bricogne, G. et al. BUSTER v.2.10.3 (Global Phasing, 2019).
Croll, T. I. ISOLDE: a physically realistic environment for model building into low-resolution electron-density maps. Acta Crystallogr. D 74, 519–530 (2018).
Google Scholar
Liebschner, D. et al. Macromolecular structure determination using X-rays, neutrons and electrons: recent developments in Phenix. Acta Crystallogr. D 75, 861–877 (2019).
Google Scholar
Hattori, M., Hibbs, R. E. & Gouaux, E. A fluorescence-detection size-exclusion chromatography-based thermostability assay for membrane protein precrystallization screening. Structure 20, 1293–1299 (2012).
Google Scholar
Alexandrov, A. I., Mileni, M., Chien, E. Y. T., Hanson, M. A. & Stevens, R. C. Microscale fluorescent thermal stability assay for membrane proteins. Structure 16, 351–359 (2008).
Google Scholar
Hawkins, P. C. D., Skillman, A. G. & Nicholls, A. Comparison of shape-matching and docking as virtual screening tools. J. Med. Chem. 50, 74–82 (2007).
Google Scholar
Molecular Operating Environment (MOE) 2019.01 (Chemical Computing Group, 2019).
Jones, G., Willett, P., Glen, R. C., Leach, A. R. & Taylor, R. Development and validation of a genetic algorithm for flexible docking. J. Mol. Biol. 267, 727–748 (1997).
Google Scholar
Mosca, R. & Schneider, T. R. RAPIDO: a web server for the alignment of protein structures in the presence of conformational changes. Nucleic Acids Res. 36, W42–W46 (2008).
Google Scholar
Caulfield, W. L. et al. The first potent and selective inhibitors of the glycine transporter type 2. J. Med. Chem. 44, 2679–2682 (2001).
Google Scholar
Madeira, F. et al. The EMBL-EBI search and sequence analysis tools APIs in 2019. Nucleic Acids Res. 47 (W1), W636–W641 (2019).
Google Scholar
Ashkenazy, H. et al. ConSurf 2016: an improved methodology to estimate and visualize evolutionary conservation in macromolecules. Nucleic Acids Res. 44 (W1), W344–W350 (2016).
Google Scholar
Winn, M. D. et al. Overview of the CCP4 suite and current developments. Acta Crystallogr. D 67, 235–242 (2011).
Google Scholar
Kantcheva, A. K. et al. Chloride binding site of neurotransmitter sodium symporters. Proc. Natl Acad. Sci. USA 110, 8489–8494 (2013).
Google Scholar
Zhang, Y.-W. et al. Chloride-dependent conformational changes in the GlyT1 glycine transporter. Proc. Natl Acad. Sci. USA (in the press) (2021).
Singh, S. K., Piscitelli, C. L., Yamashita, A. & Gouaux, E. A competitive inhibitor traps LeuT in an open-to-out conformation. Science 322, 1655–1661 (2008).
Google Scholar
Diederichs, K., & Karplus, P. A. Improved R-factors for diffraction data analysis in macromolecular crystallography. Nat. Struct. Mol. Biol. 4, 269–275 (1997).
Google Scholar
Diederichs, K., & Karplus, P. A. Linking crystallographic model and data quality. Science. 336, 1030–1033 (2012).
Google Scholar