STRUCTURE/FUNCTION ANALYSIS OF ACCHR EPITOPES

ACCHR 表位的结构/功能分析

• 批准号: 3100126
• 负责人: DAVID P RICHMAN
• 金额: $ 72.85万
• 依托单位: UNIVERSITY OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 1987
• 资助国家: 美国
• 起止时间: 1987-01-01 至 1991-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3100126
• 关键词: acetylcholine; chemical structure function

The central theme of this Program Project is to use monoclonal antibodies (mAbs) directed against defined oligopeptides within the subunits of the nicotinic acetylcholine receptor (AcChR) as site-specific probes of AcChR structure (both primary and tertiary structure) and AcChR function. Special attention will be paid to mAbs whose binding interferes with the function of this transmembrane glycoprotein neurotransmitter receptor. Projects 1-4 will assess the effects of the mAbs on particular aspects of AcChR function: Project 1 - ligand binding, Project 2 - ion fluxes by stopped flow technique; Project 3 - ion channel function by patch clamp technique; Project 4 - neuromuscular transmission in vitro and in vivo. Projects 5 and 6 will localize the epitopes of these mAbs within the AcChR structure: Project 5 - production of rationally chosen synthetic oligopeptides mimicking specific sites within AcChR subunits, assessment of the contribution of glycosyl moieties to mAb epitopes, Project 6 -identification of bound mAbs on the low resolution three-dimensional model of AcChR by both small angle X-ray diffraction and high resolution electron microscopy, as well as localization of fluorescent probes. The Scientific Core will produce new mAbs directed against the peptides produced in Project 5 and will provide purified, characterized mAbs for the various projects. In Project 4, the peptides will be used to possibly develop a new treatment modality for experimental myasthenia gravis. This Program Project represents a collaborative effort of immunologists, biochemists, electrophysiologists, peptide chemists, and physical/structural chemists in which the information from the various projects, taken together, will allow for mapping of various sites in the primary sequence of AcChR, especially functionally important sites, onto the three-dimensional model.

本项目的中心主题是使用单克隆抗体， 针对特定寡肽的抗体（mAb）， 烟碱乙酰胆碱受体（AcChR）的亚单位， AcChR结构的位点特异性探针（初级和 三级结构）和AcChR功能。 将特别注意 支付给其结合干扰本发明的功能的mAb。 跨膜糖蛋白神经递质受体 项目 1-4将评估mAb对特定方面的影响， AcChR功能：项目1 -配体结合，项目2 -离子通量 通过停流技术;项目3 -离子通道功能， 膜片钳技术;项目4 -神经肌肉传递 体外和体内。 项目5和项目6将定位 AcChR结构中的这些mAb：项目5 -生产 合理选择的合成寡肽模拟特定的 AcChR亚单位内的位点， mAb表位的糖基部分，项目6 -鉴定 低分辨率三维模型上的结合mAb AcChR的小角X射线衍射和高分辨率 电子显微镜以及荧光探针的定位。 科学核心将产生针对 在项目5中产生的肽， 用于各种项目的特征化mAb。 在项目4中， 肽将被用来开发一种新的治疗方法， 实验性重症肌无力的治疗方法。 这个程序 该项目代表了免疫学家的合作努力， 生物化学家，电生理学家，肽化学家， 物理/结构化学家，其中的信息， 各种项目结合在一起，将允许绘制各种 AcChR一级序列中的位点，特别是功能上 重要的网站，到三维模型。