STRUCTURAL AND FUNCTIONAL FEATURES OF THE 5-HT3R BINDING SITE

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 摘要：本项目的主要目标是开发基于乙酰胆碱结合蛋白(AChBP)受体蛋白的改进型微悬臂梁生物传感器。AChBP是一种可溶性蛋白质，其结构和药理与中枢和外周神经系统中存在的烟碱型乙酰胆碱受体(NAChR)惊人地相似。由于nAChR是配体门控离子通道(LGIC)超家族的成员，该项目最终将开发用于这类神经元受体的高通量药物筛选和药物发现的传感器。目标1将评估AChBP本身作为一种潜在的有用的生物传感器分子。我们将通过探索该传感器与nAChR、GABAR、GlycineR和5-HT3R配体的相互作用，初步展示该传感器在高通量药物筛选中的应用。由于AChBP通常被用作模拟配基门控离子通道受体的模板，这项工作的另一个好处是扩大了对这些重要受体的配基特异性的基础的了解。Aim 2将利用AChBP和其他受体蛋白，通过改进的连接化学方法与季博士实验室开发的改进的微悬臂阵列技术相结合，提高传感器的响应能力。

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. Abstract: The primary goal of this project is to develop improved microcantilever-based biosensors using receptor proteins based on an acetylcholine binding protein (AChBP). The AChBP is a soluble protein that displays structure and pharmacology that is strikingly similar to the nicotinic acetylcholine receptor (nAChR) present in the central and peripheral nervous systems. Since the nAChR is a member of a large super-family of ligand gated ion channels (LGICs), this project will ultimately develop sensors that will be useful in high throughput drug screening and drug discovery for this class of neuronal receptors. Aim 1 will evaluate the AChBP itself as a potentially useful biosensor molecule. We will initially demonstrate the use of this sensor in high-throughput drug screening by exploring its interaction with nAChR, GABAR, glycineR and 5-HT3R ligands. Since the AChBP is commonly used as a template for modeling of ligand gated ion channel receptors, an additional benefit of this work will be an expanded knowledge of the basis of ligand specificity at these important receptors. Aim 2 will utilize the AChBP and other receptor proteins to improve sensor response through improved conjugation chemistries in combination with improved microcantilever array technology developed in Dr. Ji's laboratory.