Subproject Investigator: Kasper B. Hansen

子项目研究员：Kasper B. Hansen

• 批准号: 10004088
• 负责人: Kasper Boe Hansen
• 金额: $ 21.75万
• 依托单位: UNIVERSITY OF MONTANA
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-01 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10004088
• 关键词: Affect; Affinity; Agonist; Allosteric Site; Antidepressive Agents; Binding; Binding Sites; Clinical Trials; Crystallization; Data; Development; Disease; Docking; Electrophysiology (science); Engineering; Genes; Glutamate Receptor; Glutamates; Glycine; Goals; Hydrophobicity; Inherited; Ion Channel; Ion Channel Gating; Kidney; Ligands; Major Depressive Disorder; Mediating; Mental disorders; Molecular Conformation; Mutagenesis; N-Methyl-D-Aspartate Receptors; N-Methylaspartate; NMDA receptor A1; Neuraxis; Patients; Pharmacology; Play; Property; Receptor Activation; Research Personnel; Resistance; Role; Site; Site-Directed Mutagenesis; Structure; Synaptic plasticity; Therapeutic; Work; X-Ray Crystallography; childhood epilepsy; clinically relevant; de novo mutation; design; disulfide bond; insight; nervous system disorder; neuron development; neurotransmission; novel; protein protein interaction; receptor function; simulation; therapeutic development; tool

NMDA-type glutamate receptors are ligand-gated ion channels that mediate excitatory neurotransmission in the central nervous system, but are also implicated in many neurological and psychiatric disorders. NMDA receptors are composed of both glycine-binding GluN1 subunits and glutamate-binding GluN2 subunits. One GluN1 subunit has been cloned and there are four types of GluN2 subunits (GluN2A-D) with different developmental and regional expression levels that endow NMDA receptors with different functional properties. GluN1 glycine site agonists show rapid onset symptomatic relief in major depressive disorder, and reduction of NMDA receptor activity by inhibiting the activity of full endogenous agonists (e.g. glycine) hold considerable promise for the development of new antidepressant drugs. In addition, recent studies have shown that up to 20% of all patients with treatment-resistant childhood epilepsy disorders may have inherited or de novo mutations in the gene encoding the GluN2A NMDA receptor subunit. New partial GluN1 agonists with selectivity between GluN2 subunits have been designed, and a unique mechanism of action have been uncovered for a class of GluN2A-selective negative allosteric modulators (NAMs) that reduce affinity of glycine binding to GluN1. Using X-ray crystallography, site-directed mutagenesis, and electrophysiological recordings of NMDA receptor function, Aim 1 of this project will investigate the binding site of GluN2A-selective NAMs and Aim 2 will define binding contacts for new classes of partial agonists at the GluN1 glycine binding site. Aim 3 will examine the allosteric interaction between NAM and glycine binding sites in more detail using site-directed mutagenesis and engineered disulfide bonds. The relationship between allosteric inhibition by GluN2A- selective NAMs and GluN1 agonist efficacy will also be uncovered. This combination of aims will uncover new regions of NMDA receptors that can be exploited for engineering of subunit-selective ligands, and provide mechanistic and structural insight to enable rational design of novel, clinically-relevant modulators that reduce GluN1 glycine site activity.

NMDA 型谷氨酸受体是配体门控离子通道，介导兴奋性神经传递 中枢神经系统，但也与许多神经和精神疾病有关。国家MDA 受体由甘氨酸结合 GluN1 亚基和谷氨酸结合 GluN2 亚基组成。一 GluN1亚基已被克隆，GluN2亚基有四种类型（GluN2A-D），各有不同 发育和区域表达水平赋予 NMDA 受体不同的功能特性。 GluN1 甘氨酸位点激动剂可快速缓解重度抑郁症的症状，并减少 通过抑制完全内源性激动剂（例如甘氨酸）的活性，NMDA 受体活性保持相当大 有望开发新的抗抑郁药物。此外，最近的研究表明，高达 所有患有难治性儿童癫痫病的患者中，20% 可能患有遗传性或新发疾病 编码 GluN2A NMDA 受体亚基的基因发生突变。新的部分 GluN1 激动剂 设计了 GluN2 亚基之间的选择性，并形成了独特的作用机制 发现一类可降低甘氨酸亲和力的 GluN2A 选择性负变构调节剂 (NAM) 与 GluN1 结合。使用 X 射线晶体学、定点诱变和电生理记录 NMDA 受体功能的研究，该项目的目标 1 将研究 GluN2A 选择性 NAM 的结合位点和 目标 2 将定义新类别的部分激动剂在 GluN1 甘氨酸结合位点的结合接触。目标 3 将使用定点更详细地检查 NAM 和甘氨酸结合位点之间的变构相互作用 诱变和工程二硫键。 GluN2A-变构抑制之间的关系 选择性 NAM 和 GluN1 激动剂的功效也将被揭示。这些目标的结合将发现新的 NMDA 受体区域可用于亚基选择性配体的工程，并提供 机制和结构洞察力能够合理设计新颖的、临床相关的调节剂，从而减少 GluN1 甘氨酸位点活性。