当前位置: X-MOL 学术ACS Chem. Neurosci. › 论文详情
Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)
Identification of Novel Allosteric Modulators of Glutamate Transporter EAAT2
ACS Chemical Neuroscience ( IF 5 ) Pub Date : 2017-11-15 00:00:00 , DOI: 10.1021/acschemneuro.7b00308
Sandhya Kortagere 1 , Ole V. Mortensen 2 , Jingsheng Xia 2 , William Lester 3 , Yuhong Fang 3 , Yellamelli Srikanth 2 , Joseph M. Salvino 2 , Andréia C. K. Fontana 2
Affiliation  

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.

中文翻译:

新型谷氨酸转运蛋白EAAT2的变构调节剂的鉴定。

兴奋性氨基酸转运蛋白(EAAT)的功能异常与多种神经系统疾病的发病机制有关,例如中风,脑外伤,癫痫和神经退行性疾病。EAAT2是负责大脑中谷氨酸清除的主要亚型,在调节传播和预防兴奋性毒性中起关键作用。因此,增加EAAT2表达或活性的化合物具有神经保护作用的治疗潜力。先前的研究确定了位于刚性三聚结构域和中央底物结合转运结构域的界面的结构域中EAAT2转运刺激的分子决定因素。在这项工作中,基于此分子域,基于杂化结构的方法被应用于创建高分辨率药效基团。随后,对小分子文库进行了虚拟筛选,确定了在拟议域中相互作用的10个命中分子。其中,确定了三种化合物为激活剂,四种为抑制剂,三种对EAAT2介导的转运无影响体外。进一步证明了两种最上等的EAAT2激活剂的功效,效能和对谷氨酸的选择性,优于单胺转运蛋白亚型和NMDA受体,以及在培养的星形胶质细胞中的功效。诱变研究表明,EAAT2激活剂与形成三聚化域和转运域之间界面的残基相互作用。这些化合物提高了谷氨酸的转运速率,而对底物的相互作用没有影响,提示了一种变构机制。这些新颖的EAAT2阳性正构构调节剂的鉴定为基于谷氨酸转运增强的疗法的开发提供了创新的方法。
更新日期:2017-11-15
down
wechat
bug