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Identification of Novel Allosteric Modulators of Glutamate Transporter EAAT2
Journal article   Peer reviewed

Identification of Novel Allosteric Modulators of Glutamate Transporter EAAT2

Sandhya Kortagere, Ole V Mortensen, Jingsheng Xia, William Lester, Yuhong Fang, Yellamelli Srikanth, Joseph M Salvino and Andréia C K Fontana
ACS chemical neuroscience, v 9(3), pp 522-534
21 Mar 2018
PMID: 29140675

Abstract

hybrid structure based method EAAT2 activators glutamate transporter transport enhancement positive allosteric modulation glutamate uptake EAAT2
Dysfunction of excitatory amino acid transporters (EAATs) has been implicated in the pathogenesis of various neurological disorders, such as stroke, brain trauma, epilepsy, and neurodegenerative diseases, among others. EAAT2 is the main subtype responsible for glutamate clearance in the brain, having a key role in regulating transmission and preventing excitotoxicity. Therefore, compounds that increase the expression or activity of EAAT2 have therapeutic potential for neuroprotection. Previous studies identified molecular determinants for EAAT2 transport stimulation in a structural domain that lies at the interface of the rigid trimerization domain and the central substrate binding transport domain. In this work, a hybrid structure based approach was applied, based on this molecular domain, to create a high-resolution pharmacophore. Subsequently, virtual screening of a library of small molecules was performed, identifying 10 hit molecules that interact at the proposed domain. Among these, three compounds were determined to be activators, four were inhibitors, and three had no effect on EAAT2-mediated transport in vitro. Further characterization of the two best ranking EAAT2 activators for efficacy, potency, and selectivity for glutamate over monoamine transporters subtypes and NMDA receptors and for efficacy in cultured astrocytes is demonstrated. Mutagenesis studies suggest that the EAAT2 activators interact with residues forming the interface between the trimerization and transport domains. These compounds enhance the glutamate translocation rate, with no effect on substrate interaction, suggesting an allosteric mechanism. The identification of these novel positive allosteric modulators of EAAT2 offers an innovative approach for the development of therapies based on glutamate transport enhancement.

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Collaboration types
Domestic collaboration
Web of Science research areas
Biochemistry & Molecular Biology
Chemistry, Medicinal
Neurosciences
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