PROJECT 1 - MOLECULAR ANALYSIS OF NMDA RECEPTOR MODULATORY SITES

项目 1 - NMDA 受体调节位点的分子分析

• 批准号: 8535874
• 负责人: STUART A LIPTON
• 金额: $ 6.83万
• 依托单位: SANFORD BURNHAM PREBYS MEDICAL DISCOVERY INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-03-01 至 2017-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8535874
• 关键词: Abate; Active Sites; Affect; Affinity; Alzheimer' s Disease; Binding; Biological Assay; Brain; Brain Hypoxia-Ischemia; Brain Injuries; Calcium; Chemicals; Cognitive deficits; Crystallization; Cysteine; Developmental Disabilities; Down Syndrome; Electrophysiology (science); Etiology; Family; Functional disorder; Genetic; Glutamate Receptor; Grant; Impaired cognition; In Vitro; Instruction; Intellectual functioning disability; Ion Channel; Ischemic-Hypoxic Encephalopathy; Knowledge; Ligand Binding Domain; Ligands; Magnesium; Mediating; Memantine; Modification; Molecular; Molecular Analysis; N-Methyl-D-Aspartate Receptors; N-Methylaspartate; NMDA receptor antagonist; NR1 gene; National Research Service Awards; Neuraxis; Neurons; Neuroprotective Agents; Oxidation-Reduction; Peptides; Perinatal Hypoxia; Permeability; Pharmaceutical Preparations; Principal Investigator; Property; Reaction; Research Personnel; Role; Screening procedure; Single Nucleotide Polymorphism; Site; Site-Directed Mutagenesis; Structure; Sulfhydryl Compounds; Synapses; Synaptic Receptors; Testing; Therapeutic; Vertebral column; Vestibule; base; design; developmental disease; gene therapy; high throughput screening; improved; in vivo; nervous system disorder; neuroprotection; novel; prevent; programs; protein function; receptor

Project I - Zhang & Chen Program Director/Principal Investigator (Last, First, Middle): Lipton, Stuart A. PROJECT SUMMARY (See instructions): The N-methyl-D-aspartate subtype of gluatmate receptor (NMDAR) is essential for normal function of the central nervous system (CNS). However, excessive activation of NMDARs, particulariy of extrasynaptic as opposed to synaptic receptors, mediates, at least in part, neuronal or synaptic damage in many neurological disorders, such as hypoxic-ischemic brain injury and, as recently suggested, in Down syndrome. Blockade of excessive NMDAR activity must be achieved without interference with its normal brain function. We have taken two approaches for clinically-tolerated pharmacological and genetic intervention on NMDARs. One approach is to use Memantine but also NO species to further down regulate the NMDAR by S-nitrosylation. The structural determinants on NMDARs for the action of Memantine and NO-like species will be characterized further under the auspices of this grant. Another approach is to utilize the inhibitory effect of a novel family of NMDAR subunits, composed of NRSA and NRSB, to downregulate NMDARs by affecting channel permeability, in a sense mimicking the effect of the NMDAR antagonist drugs that are also being developed here. We will study the role of the MS domain of NRS subunits that downregulate activity of NMDARs and also design NRS ligand-binding domain (LBD)-based screening assays to discover new compounds that modulate NRS-containing receptors. These agents will be useful for characterizing NRS-containing receptors, and possibly for neuroprotection. Accordingly, the Specific Aims of this proposal are as follows: 1) To study the effect of S-nitrosylation/redox modulation of the loose linker region between the amino-terminal domain (ATD) and the LBD of NMDARs by electrophysiology; 2) To develop LBD-derived screening assays to screen for ligands selective for the NRS subunit of the NMDAR. These ligands will be further characterized and refined by secondary assays, chemical modification, and co-crystallization; 3) To study the inhibitory effect of peptides derived from the out vestibule (MS) region of NRS subunits on NMDAR permeability.

项目 I - 张和陈 项目总监/首席研究员（最后、第一、中间）：Lipton, Stuart A. 项目摘要（参见说明）： 谷氨酸受体 (NMDAR) 的 N-甲基-D-天冬氨酸亚型对于中枢神经系统 (CNS) 的正常功能至关重要。然而，NMDAR 的过度激活，特别是突触外而非突触受体的过度激活，至少部分介导许多神经系统疾病中的神经元或突触损伤，例如缺氧缺血性脑损伤，以及最近提出的唐氏综合症。阻断过度的 NMDAR 活性必须在不干扰其正常大脑功能的情况下实现。我们采取了两种方法对 NMDAR 进行临床耐受的药理学和遗传干预。一种方法是使用美金刚和 NO 物质通过 S-亚硝基化进一步下调 NMDAR。 在这项资助的支持下，将进一步表征 NMDAR 对美金刚和 NO 样物质作用的结构决定因素。另一种方法是利用由 NRSA 和 NRSB 组成的新型 NMDAR 亚基家族的抑制作用，通过影响通道通透性来下调 NMDAR，在某种意义上模仿也在开发的 NMDAR 拮抗剂药物的作用。我们将研究 NRS 亚基的 MS 结构域在下调 NMDAR 活性方面​​的作用，并设计基于 NRS 配体结合结构域 (LBD) 的筛选分析，以发现调节含 NRS 受体的新化合物。这些试剂可用于表征包含 NRS 的受体，并可能用于神经保护。 因此，本提案的具体目标如下： 1）通过电生理学研究NMDAR的氨基末端结构域（ATD）和LBD之间的松散连接区的S-亚硝基化/氧化还原调节的影响； 2) 开发LBD衍生的筛选方法来筛选对NMDAR的NRS亚基具有选择性的配体。这些配体将通过二次测定、化学修饰和共结晶进一步表征和精制； 3) 研究NRS亚基前庭外(MS)区衍生的肽对NMDAR通透性的抑制作用。