A Computational Approach Yields Selective Inhibitors of Human Excitatory Amino Acid Transporter 2 (EAAT2)

Overview

Journal J Biol Chem

Specialty Biochemistry

Date 2020 Feb 22

PMID 32079674

Citations 2

Authors

Kelly L Damm-Ganamet

Marie-Laure Rives

Alan D Wickenden

Heather M McAllister

Taraneh Mirzadegan

Affiliations

Soon will be listed here.

Abstract

Excitatory amino acid transporters (EAATs) represent a protein family that is an emerging drug target with great therapeutic potential for managing central nervous system disorders characterized by dysregulation of glutamatergic neurotransmission. As such, it is of significant interest to discover selective modulators of EAAT2 function. Here, we applied computational methods to identify specific EAAT2 inhibitors. Utilizing a homology model of human EAAT2, we identified a binding pocket at the interface of the transport and trimerization domain. We next conducted a high-throughput virtual screen against this site and identified a selective class of EAAT2 inhibitors that were tested in glutamate uptake and whole-cell electrophysiology assays. These compounds represent potentially useful pharmacological tools suitable for further exploration of the therapeutic potential of EAAT2 and may provide molecular insights into mechanisms of allosteric modulation for glutamate transporters.

Citing Articles

A Medicinal Chemistry Perspective on Excitatory Amino Acid Transporter 2 Dysfunction in Neurodegenerative Diseases.

Fontana I, Souza D, Souza D, Gee A, Zimmer E, Bongarzone S J Med Chem. 2023; 66(4):2330-2346.

PMID: 36787643 PMC: 9969404. DOI: 10.1021/acs.jmedchem.2c01572.

Peripheral and Central Glutamate Dyshomeostasis in Neurodegenerative Disorders.

Onaolapo A, Onaolapo O Curr Neuropharmacol. 2020; 19(7):1069-1089.

PMID: 33059576 PMC: 8686318. DOI: 10.2174/1570159X18666201015161919.

References

Baell J, Holloway G . New substructure filters for removal of pan assay interference compounds (PAINS) from screening libraries and for their exclusion in bioassays. J Med Chem. 2010; 53(7):2719-40. DOI: 10.1021/jm901137j. View

Lounnas V, Ritschel T, Kelder J, McGuire R, Bywater R, Foloppe N . Current progress in Structure-Based Rational Drug Design marks a new mindset in drug discovery. Comput Struct Biotechnol J. 2014; 5:e201302011. PMC: 3962124. DOI: 10.5936/csbj.201302011. View

Veljkovic V, Loiseau P, Figadere B, Glisic S, Veljkovic N, Perovic V . Virtual screen for repurposing approved and experimental drugs for candidate inhibitors of EBOLA virus infection. F1000Res. 2015; 4:34. PMC: 4329668. DOI: 10.12688/f1000research.6110.1. View

Yernool D, Boudker O, Jin Y, Gouaux E . Structure of a glutamate transporter homologue from Pyrococcus horikoshii. Nature. 2004; 431(7010):811-8. DOI: 10.1038/nature03018. View

Lavecchia A, Di Giovanni C . Virtual screening strategies in drug discovery: a critical review. Curr Med Chem. 2013; 20(23):2839-60. DOI: 10.2174/09298673113209990001. View

Falcucci R, Wertz R, Green J, Meucci O, Salvino J, Fontana A . Novel Positive Allosteric Modulators of Glutamate Transport Have Neuroprotective Properties in an in Vitro Excitotoxic Model. ACS Chem Neurosci. 2019; 10(8):3437-3453. PMC: 7099866. DOI: 10.1021/acschemneuro.9b00061. View

Fairman W, Vandenberg R, Arriza J, Kavanaugh M, Amara S . An excitatory amino-acid transporter with properties of a ligand-gated chloride channel. Nature. 1995; 375(6532):599-603. DOI: 10.1038/375599a0. View

Rettenmaier T, Fan H, Karpiak J, Doak A, Sali A, Shoichet B . Small-Molecule Allosteric Modulators of the Protein Kinase PDK1 from Structure-Based Docking. J Med Chem. 2015; 58(20):8285-8291. PMC: 5099076. DOI: 10.1021/acs.jmedchem.5b01216. View

Sliwoski G, Kothiwale S, Meiler J, Lowe Jr E . Computational methods in drug discovery. Pharmacol Rev. 2014; 66(1):334-95. PMC: 3880464. DOI: 10.1124/pr.112.007336. View

10.

Rajkowska G, Stockmeier C . Astrocyte pathology in major depressive disorder: insights from human postmortem brain tissue. Curr Drug Targets. 2013; 14(11):1225-36. PMC: 3799810. DOI: 10.2174/13894501113149990156. View

11.

Soga S, Shirai H, Kobori M, Hirayama N . Use of amino acid composition to predict ligand-binding sites. J Chem Inf Model. 2007; 47(2):400-6. DOI: 10.1021/ci6002202. View

12.

Halgren T . Identifying and characterizing binding sites and assessing druggability. J Chem Inf Model. 2009; 49(2):377-89. DOI: 10.1021/ci800324m. View

13.

Nakagawa T, Kaneko S . SLC1 glutamate transporters and diseases: psychiatric diseases and pathological pain. Curr Mol Pharmacol. 2013; 6(2):66-73. DOI: 10.2174/18744672113069990033. View

14.

Verdon G, Oh S, Serio R, Boudker O . Coupled ion binding and structural transitions along the transport cycle of glutamate transporters. Elife. 2014; 3:e02283. PMC: 4051121. DOI: 10.7554/eLife.02283. View

15.

Hack M, Rassokhin D, Buyck C, Seierstad M, Skalkin A, ten Holte P . Library enhancement through the wisdom of crowds. J Chem Inf Model. 2011; 51(12):3275-86. DOI: 10.1021/ci200446y. View

16.

Behrens P, Franz P, Woodman B, Lindenberg K, Landwehrmeyer G . Impaired glutamate transport and glutamate-glutamine cycling: downstream effects of the Huntington mutation. Brain. 2002; 125(Pt 8):1908-22. DOI: 10.1093/brain/awf180. View

17.

Berman H, Westbrook J, Feng Z, Gilliland G, Bhat T, Weissig H . The Protein Data Bank. Nucleic Acids Res. 1999; 28(1):235-42. PMC: 102472. DOI: 10.1093/nar/28.1.235. View

18.

Abdallah C, Averill C, Salas R, Averill L, Baldwin P, Krystal J . Prefrontal Connectivity and Glutamate Transmission: Relevance to Depression Pathophysiology and Ketamine Treatment. Biol Psychiatry Cogn Neurosci Neuroimaging. 2017; 2(7):566-574. PMC: 5635826. DOI: 10.1016/j.bpsc.2017.04.006. View

19.

Fontana A, de Oliveira Beleboni R, Wojewodzic M, dos Santos W, Coutinho-Netto J, Grutle N . Enhancing glutamate transport: mechanism of action of Parawixin1, a neuroprotective compound from Parawixia bistriata spider venom. Mol Pharmacol. 2007; 72(5):1228-37. DOI: 10.1124/mol.107.037127. View

20.

Friesner R, Banks J, Murphy R, Halgren T, Klicic J, Mainz D . Glide: a new approach for rapid, accurate docking and scoring. 1. Method and assessment of docking accuracy. J Med Chem. 2004; 47(7):1739-49. DOI: 10.1021/jm0306430. View