Structure and Function of NMDA Receptors

NMDA 受体的结构和功能

• 批准号: 9232209
• 负责人: Hiroyasu Furukawa
• 金额: $ 49.89万
• 依托单位: COLD SPRING HARBOR LABORATORY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-03-01 至 2020-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9232209
• 关键词: Address; Affinity; Agonist; Allosteric Site; Alzheimer' s Disease; Antibodies; Binding; Binding Sites; Biochemistry; Brain; Brain Diseases; Brain region; Chemical Structure; Chemicals; Complex; Coupled; Development; Disease; Electrophysiology (science); Elements; Exhibits; Extracellular Domain; Family; Fc Receptor; Funding; Gated Ion Channel; Glutamate Receptor; Glutamates; Glycine; Goals; Ion Channel; Ion Channel Gating; Knowledge; Ligand Binding; Ligand Binding Domain; Ligands; Magnesium; Mammals; Mediating; Mental Depression; Mental Health; Molecular; Molecular Profiling; N-Methyl-D-Aspartate Receptors; NMDA receptor A1; Neurologic; Neurons; Parkinson Disease; Pattern; Pharmacology; Play; Property; Public Health; RNA Splicing; Reagent; Regulation; Research; Resolution; Role; Schizophrenia; Site-Directed Mutagenesis; Specificity; Structure; Synaptic Transmission; Testing; Therapeutic; Therapeutic Uses; Therapeutic antibodies; Transmembrane Domain; Variant; X-Ray Crystallography; Zinc; base; experimental study; ifenprodil; inhibitor/antagonist; insight; member; mental health related disorder; nervous system disorder; novel; novel strategies; public health relevance; receptor

DESCRIPTION (provided by applicant): The goal of this project is to define the molecular mechanism underlying subtype-specificity in N-methyl-D- aspartate receptors (NMDARs) to facilitate development of subtype-specific reagents for controlling NMDAR activities. NMDARs belong to the family of ionotropic glutamate receptors, which mediate the majority of fast excitatory synaptic transmission in mammalian brains. Abnormal activity of NMDARs is implicated in various neurological disorders and diseases including schizophrenia, depression, Alzheimer's disease, and Parkinson's disease. Those receptors are multimeric ligand-gated ion channels composed mainly of GluN1 and GluN2 subunits that bind to glycine and L-glutamate at the extracellular domain, respectively. Gating of transmembrane ion channels is mediated by concurrent binding of glycine and L-glutamate to the ligand- binding domain (LBD) and is allosterically regulated by binding of modulator compounds including phenylethanolamines and Zn2+ to the amino terminal domain (ATD). Importantly, functional properties of NMDARs subtypes, which are defined by four distinct GluN2 subunits (A though D), exhibit dramatically different functional properties. Different NMDAR subtypes are expressed in discrete regions of the brain at given developmental stages and are also associated with distinct neurological diseases and disorders. Thus, understanding the molecular basis for subtype-specificity will be necessary in order to develop specific reagents for treatment of the above neurological diseases. Despite much enthusiasm, the field is limited to one useful subtype-specific compound, phenylethanolamine, which targets GluN1/GluN2B NMDARs but is associated with off-target effects when used therapeutically. Development of reagents targeting other subtypes such as GluN1/GluN2A has been hampered due to limited amount of structural information on subtypes of NMDARs, which would allow comprehensive structural comparison. To obtain a mechanistic understanding of subtype-specificity in NMDARs and to facilitate development of subtype-specific reagents for GluN1/GluN2A and GluN1/GluN2B NMDARs, we will conduct research aimed at: Aim 1 obtaining an in-depth understanding of the ligand-binding site in GluN1/GluN2B ATD and GluN1/GluN2A LBD; Aim 2 defining the molecular mechanism of subtype-specific allosteric inhibition in GluN1/GluN2A ATD; and Aim 3 determining the binding and inhibition mechanism of GluN1/GluN2B NMDAR by inhibitory antibody that we recently developed. These three goals will be achieved by obtaining the structural information of ATD and LBD and testing structure- based hypotheses by electrophysiology. Successful completion of the proposed studies will provide unprecedented insights into ligand-binding sites in ATD, LBD, and molecular elements underlying subtype- specificity, and to demonstrate a novel approach to inhibit NMDAR in a subtype-specific manner using inhibitory antibodies. These findings will facilitate development of subtype-specific reagents to study and treat the mental health related disorders above.

描述（由申请人提供）：该项目的目的是定义N-甲基-D-天冬氨酸受体（NMDARS）中亚型的分子机制，以促进用于控制NMDAR活动的亚型特异性试剂的发展。 NMDAR属于离子型谷氨酸受体家族，它介导了哺乳动物大脑中大多数快速兴奋性突触传播。 NMDAR的异常活性与各种神经系统疾病和疾病有关，包括精神分裂症，抑郁症，阿尔茨海默氏病和帕金森氏病。这些受体是多聚体门控离子通道，主要由Glun1和Glun2亚基组成，分别与甘氨酸和L-谷氨酸结合，分别在细胞外结构域结合。跨膜离子通道的输卵是通过甘氨酸和l-谷氨酸与配体结合结构域（LBD）的同时结合介导的，并通过调节化合物的结合（包括苯甲醇胺和Zn2+）与氨基末端（ATD）的结合来构层调节。重要的是，由四个不同的glun2亚基（a a d）定义的NMDARS亚型的功能特性表现出截然不同的功能性能。不同的NMDAR亚型在给定发育阶段在大脑的离散区域中表示，也与不同的神经系统疾病和疾病有关。因此，必须了解亚型特异性的分子基础，以开发特定的试剂来治疗上述神经系统疾病。尽管热情洋溢，但该领域仍限于一种有用的亚型特异性化合物苯甲醇胺，该化合物靶向GLUN1/GLUN2B NMDAR，但在治疗时使用时与脱靶效应相关。靶向其他亚型（例如glun1/glun2a）的试剂的开发受到了阻碍，这是由于NMDARS亚型的结构性信息有限，这将允许全面的结构比较。 To obtain a mechanistic understanding of subtype-specificity in NMDARs and to facilitate development of subtype-specific reagents for GluN1/GluN2A and GluN1/GluN2B NMDARs, we will conduct research aimed at: Aim 1 obtaining an in-depth understanding of the ligand-binding site in GluN1/GluN2B ATD and GluN1/GluN2A LBD; AIM 2定义Glun1/Glun2a ATD中亚型特异性变构抑制的分子机制；目标3通过我们最近开发的抑制性抗体来确定Glun1/Glun2b NMDAR的结合和抑制机制。这三个目标将通过获得ATD和LBD的结构信息以及通过电生理学的基于测试结构的假设来实现。拟议的研究的成功完成将提供前所未有的见解，对亚型亚型的ATD，LBD和分子元素中的配体结合位点，并使用抑制性抗体证明了一种新型的NMDAR抑制NMDAR的方法。这些发现将有助于开发亚型特异性试剂，以研究和治疗上面的心理健康相关疾病。