Structure and Allosteric Inhibition of Excitatory Amino Acid Transporter 1

Overview

Journal Nature

Specialty Science

Date 2017 Apr 21

PMID 28424515

Citations 116

Authors

Juan C Canul-Tec

Reda Assal

Erica Cirri

Pierre Legrand

Sebastien Brier

Julia Chamot-Rooke

Nicolas Reyes

Affiliations

Soon will be listed here.

Abstract

Human members of the solute carrier 1 (SLC1) family of transporters take up excitatory neurotransmitters in the brain and amino acids in peripheral organs. Dysregulation of the function of SLC1 transporters is associated with neurodegenerative disorders and cancer. Here we present crystal structures of a thermostabilized human SLC1 transporter, the excitatory amino acid transporter 1 (EAAT1), with and without allosteric and competitive inhibitors bound. The structures reveal architectural features of the human transporters, such as intra- and extracellular domains that have potential roles in transport function, regulation by lipids and post-translational modifications. The coordination of the allosteric inhibitor in the structures and the change in the transporter dynamics measured by hydrogen-deuterium exchange mass spectrometry reveal a mechanism of inhibition, in which the transporter is locked in the outward-facing states of the transport cycle. Our results provide insights into the molecular mechanisms underlying the function and pharmacology of human SLC1 transporters.

Citing Articles

Inhibitory Potential of the Truncated Isoforms on Glutamate Transporter Oligomerization Identified by Computational Analysis of Gene-Centric Isoform Maps.

Karagol A, Karagol T, Li M, Zhang S Pharm Res. 2024; 41(11):2173-2187.

PMID: 39487385 PMC: 11599315. DOI: 10.1007/s11095-024-03786-z.

Loss of Carbamoyl Phosphate Synthetase 1 Potentiates Hepatocellular Carcinoma Metastasis by Reducing Aspartate Level.

Chen S, Tang Q, Hu M, Song S, Wu X, Zhou Y Adv Sci (Weinh). 2024; 11(45):e2402703.

PMID: 39387452 PMC: 11615744. DOI: 10.1002/advs.202402703.

Succinate Dehydrogenase and Human Disease: Novel Insights into a Well-Known Enzyme.

Esteban-Amo M, Jimenez-Cuadrado P, Serrano-Lorenzo P, de la Fuente M, Simarro M Biomedicines. 2024; 12(9).

PMID: 39335562 PMC: 11429145. DOI: 10.3390/biomedicines12092050.

Molecular Dynamic Simulations Reveal that Water-Soluble QTY-Variants of Glutamate Transporters EAA1, EAA2 and EAA3 Retain the Conformational Characteristics of Native Transporters.

Karagol A, Karagol T, Zhang S Pharm Res. 2024; 41(10):1965-1977.

PMID: 39322794 PMC: 11530497. DOI: 10.1007/s11095-024-03769-0.

Structural basis of the excitatory amino acid transporter 3 substrate recognition.

Qiu B, Boudker O bioRxiv. 2024; .

PMID: 39282329 PMC: 11398500. DOI: 10.1101/2024.09.05.611541.

References

Choi K, Jen J, Choi S, Shin J, Kim H, Kim H . Late-onset episodic ataxia associated with SLC1A3 mutation. J Hum Genet. 2016; 62(3):443-446. DOI: 10.1038/jhg.2016.137. View

Zerangue N, Kavanaugh M . Flux coupling in a neuronal glutamate transporter. Nature. 1996; 383(6601):634-7. DOI: 10.1038/383634a0. View

Rothstein J, van Kammen M, Levey A, Martin L, Kuncl R . Selective loss of glial glutamate transporter GLT-1 in amyotrophic lateral sclerosis. Ann Neurol. 1995; 38(1):73-84. DOI: 10.1002/ana.410380114. View

Waterhouse A, Procter J, Martin D, Clamp M, Barton G . Jalview Version 2--a multiple sequence alignment editor and analysis workbench. Bioinformatics. 2009; 25(9):1189-91. PMC: 2672624. DOI: 10.1093/bioinformatics/btp033. View

Emsley P, Lohkamp B, Scott W, Cowtan K . Features and development of Coot. Acta Crystallogr D Biol Crystallogr. 2010; 66(Pt 4):486-501. PMC: 2852313. DOI: 10.1107/S0907444910007493. View

Seal R, Amara S . A reentrant loop domain in the glutamate carrier EAAT1 participates in substrate binding and translocation. Neuron. 1999; 21(6):1487-98. DOI: 10.1016/s0896-6273(00)80666-2. View

Evans P, Murshudov G . How good are my data and what is the resolution?. Acta Crystallogr D Biol Crystallogr. 2013; 69(Pt 7):1204-14. PMC: 3689523. DOI: 10.1107/S0907444913000061. View

Focke P, Moenne-Loccoz P, Larsson H . Opposite movement of the external gate of a glutamate transporter homolog upon binding cotransported sodium compared with substrate. J Neurosci. 2011; 31(16):6255-62. PMC: 3096012. DOI: 10.1523/JNEUROSCI.6096-10.2011. View

Pilc A, Wieronska J, Skolnick P . Glutamate-based antidepressants: preclinical psychopharmacology. Biol Psychiatry. 2013; 73(12):1125-32. DOI: 10.1016/j.biopsych.2013.01.021. View

10.

McCoy A, Grosse-Kunstleve R, Adams P, Winn M, Storoni L, Read R . Phaser crystallographic software. J Appl Crystallogr. 2009; 40(Pt 4):658-674. PMC: 2483472. DOI: 10.1107/S0021889807021206. View

11.

Guskov A, Jensen S, Faustino I, Marrink S, Slotboom D . Coupled binding mechanism of three sodium ions and aspartate in the glutamate transporter homologue Glt. Nat Commun. 2016; 7:13420. PMC: 5110648. DOI: 10.1038/ncomms13420. View

12.

Danbolt N . Glutamate uptake. Prog Neurobiol. 2001; 65(1):1-105. DOI: 10.1016/s0301-0082(00)00067-8. View

13.

Butchbach M, Tian G, Guo H, Lin C . Association of excitatory amino acid transporters, especially EAAT2, with cholesterol-rich lipid raft microdomains: importance for excitatory amino acid transporter localization and function. J Biol Chem. 2004; 279(33):34388-96. DOI: 10.1074/jbc.M403938200. View

14.

Shimizu K, Kaira K, Tomizawa Y, Sunaga N, Kawashima O, Oriuchi N . ASC amino-acid transporter 2 (ASCT2) as a novel prognostic marker in non-small cell lung cancer. Br J Cancer. 2014; 110(8):2030-9. PMC: 3992511. DOI: 10.1038/bjc.2014.88. View

15.

Wang Q, Hardie R, Hoy A, van Geldermalsen M, Gao D, Fazli L . Targeting ASCT2-mediated glutamine uptake blocks prostate cancer growth and tumour development. J Pathol. 2015; 236(3):278-89. PMC: 4973854. DOI: 10.1002/path.4518. View

16.

Zerangue N, Kavanaugh M . ASCT-1 is a neutral amino acid exchanger with chloride channel activity. J Biol Chem. 1996; 271(45):27991-4. DOI: 10.1074/jbc.271.45.27991. View

17.

Grunewald M, Bendahan A, Kanner B . Biotinylation of single cysteine mutants of the glutamate transporter GLT-1 from rat brain reveals its unusual topology. Neuron. 1998; 21(3):623-32. DOI: 10.1016/s0896-6273(00)80572-3. View

18.

Watzke N, Grewer C . The anion conductance of the glutamate transporter EAAC1 depends on the direction of glutamate transport. FEBS Lett. 2001; 503(2-3):121-5. DOI: 10.1016/s0014-5793(01)02715-6. View

19.

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

20.

Tao Z, Rosental N, Kanner B, Gameiro A, Mwaura J, Grewer C . Mechanism of cation binding to the glutamate transporter EAAC1 probed with mutation of the conserved amino acid residue Thr101. J Biol Chem. 2010; 285(23):17725-33. PMC: 2878536. DOI: 10.1074/jbc.M110.121798. View