ALLOSTERIC POTENTIATION OF NEURONAL NICOTINIC ACETYLCHOLINE RECEPTORS

神经元烟碱乙酰胆碱受体的变构增强

• 批准号: 7960096
• 负责人: BRIAN WILLIAM EDMONDS
• 金额: $ 8.89万
• 依托单位: UNIVERSITY OF ALASKA FAIRBANKS
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-05-01 至 2010-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7960096
• 关键词: Address; Agonist; Alaska; Allosteric Site; Binding; Binding Sites; Cells; Chemicals; Chemosensitization; Cholinergic Receptors; Classification; Computer Retrieval of Information on Scientific Projects Database; Disease; Extracellular Domain; Faculty; Funding; Grant; Harvest; Homology Modeling; Institution; Methodology; Methods; Molecular; Mutagenesis; Neurons; Nicotinic Receptors; Oocytes; Physiologic pulse; Process; Property; Recruitment Activity; Research; Research Personnel; Research Proposals; Resources; Source; Students; System; United States National Institutes of Health; Work; analog; design; environmental agent; graduate student; improved; outreach; patch clamp; receptor; response

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Dr. Edmonds joined the INBRE support faculty through the outreach core in 2006-2007. He has successfully recruited a graduate student that is officially a doctoral student of the Fairbanks campus however is working now in the Dr Edmonds lab on the Juneau campus. Defined research proposal from Dr. Edmonds. Aim 1: Identify the binding site of the selective allosteric potentiator deformylflustrabromine on the a4b2 subtype of the acetylcholine receptor. Homology modeling of the extracellular domain of the receptor will be used to identify candidate binding sites for dFBr. Identification of the binding site(s) will be confirmed by examining the functional effects of systematic mutagenesis of residues in the vicinity of the putative binding site. Aim 2: Identify the molecular features of deformylflustrabromine critical for binding and activity at the allosteric site. This aim will be addressed using standard electrophysiological methods to screen synthetic analogs of naturally occurring dFBr. Compounds that potentiate the action of ACh will be investigated further in Aim 3. The structural and chemical features of active compounds will be noted and used in the design process to improve functional properties of the compound(s). Aim 3: Determine the mechanism of potentiation for the deformylflustrabromine analogs identified in Research Aim 2. For this aim we will utilize patch clamp methodology in combination with a fast agonist application system to examine the effects of dFBr analog(s) (Aim 2) on single receptor-channel responses to pulses of ACh. a4b2 receptors will be harvested either from oocytes (transient expression) or stably transfected HEK cells.

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。列出的机构为 中心，但不一定是研究者所在的机构。 博士埃德蒙兹在2006-2007年通过外展核心加入了INBRE支持学院。 他已经成功地招募了一名研究生，这名研究生正式成为费尔班克斯校区的博士生，但现在正在朱诺校区的埃德蒙兹博士实验室工作。 Edmonds博士的研究计划。 目的1：确定选择性别构增效剂去甲氟溴铵在乙酰胆碱受体a4 b2亚型上的结合位点。 受体胞外结构域的同源性建模将用于鉴定dFBr的候选结合位点。 通过检查推定结合位点附近残基系统性诱变的功能效应，确认结合位点的鉴别。 目的2：确定在变构位点的结合和活性的关键去甲酰基氟曲溴的分子特征。 这一目标将使用标准的电生理方法来筛选天然存在的dFBr的合成类似物。 增强乙酰胆碱作用的化合物将在目标3中进一步研究。 活性化合物的结构和化学特征将被记录并用于设计过程中，以改善化合物的功能特性。 目的3：确定研究目的2中鉴定的去甲酰基氟溴环类似物的增效机制。 为此，我们将利用膜片钳方法结合快速激动剂应用系统来检查dFBr类似物（目的2）对ACh脉冲的单受体通道反应的影响。 从卵母细胞（瞬时表达）或稳定转染的HEK细胞收获α 4 b2受体。