STRUCTURE AND FUNCTION OF THE ACETYLCHOLINE RECEPTOR

乙酰胆碱受体的结构和功能

• 批准号: 3075215
• 负责人: STEEN E PEDERSEN
• 金额: $ 6.68万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-12-01 至 1997-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3075215
• 关键词: Torpedo; acetylcholine; affinity labeling; alternatives to animals in research; animal poison; calcium channel blockers; chemical binding; crosslink; ethidium; fluorescence spectrometry; gel filtration chromatography; high performance liquid chromatography; laboratory mouse; molecular site; neurotoxins; nicotinic receptors; protein sequence; protein structure function; tubocurarine; western blottings

The objectives of the research application are to understand the structural features of the nicotinic acetylcholine receptor (AChR) and to relate these structures to the function of the receptor. The structure of the acetylcholine binding sites will be investigated by photoaffinity labelling to determine respective contributions of the subunits to the binding sites. Photoaffinity labels derivatized from d- tubocurarine, ethidium bromide and conotoxins will be designed to probe the contributions gamma- and delta-subunits to the agonist binding sites. The topology of the AChR primary subunit sequence will be analyzed by labelling of protein exposed on the extracellular surface of the AChR. The labelled sites will be identified by isolation of labelled fragments and identification of labelled residues. The location of putative intracellularly exposed fragment a156-179 will specifically be evaluated. Chemical crosslinking will be used to define near-neighbor relationships of the subunits and of individual amino acids within subunits.

研究申请的目标是了解 烟碱乙酰胆碱受体（ACHR）和 将这些结构与受体的功能联系起来。 这 乙酰胆碱结合位点的结构将通过 光性标签以确定相应的贡献 绑定位点的亚基。 从d-衍生的光性标签 将设计用于探测 对激动剂结合位点的伽马和三角洲贡献的贡献。 ACHR主要亚基序列的拓扑将通过 暴露在ACHR的细胞外表面上的蛋白质的标记。 标记的位点将通过隔离标记的片段来识别 并识别标记的残基。 推定的位置 细胞内暴露的片段A156-179将被专门评估。 化学交联将用于定义接近邻近的关系 亚基中的亚基和单个氨基酸的。