KINETICS OF M1 MUSCARINIC RECEPTOR EXPRESSION ON THE PLASMA MEMBRANE

质膜上 M1 毒蕈碱受体表达的动力学

• 批准号: 8170985
• 负责人: Fred J Ehlert
• 金额: $ 0.07万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-08-01 至 2011-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8170985
• 关键词: Binding; Cell fusion; Cell membrane; Chimeric Proteins; Chinese Hamster Ovary Cell; Cleaved cell; Computer Retrieval of Information on Scientific Projects Database; Dissociation; Funding; Grant; Green Fluorescent Proteins; Human; Institution; Kinetics; Ligands; Muscarinic Acetylcholine Receptor; Muscarinic M1 Receptor; Research; Research Personnel; Resources; Source; Travel; United States National Institutes of Health; protein aggregate; receptor expression

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. We plan to study the kinetics of the expression of the M1 muscarinic receptor on the plasma membrane of Chinese hamster ovary (CHO) cells using a fusion protein consisting of a conditional aggregation domain (CAD) fused to the N-terminus of the human M1 receptor and green fluorescent protein (GFP) fused to the C-terminus. Following transient expression in CHO cells, the fusion protein (GFP-M1-CAD) is trapped in the ER, but can be induced to exit following application of a ligand (AP21998) that binds to the CAD and causes dissociation of fusion protein aggregates in the ER. After exiting the ER, the CAD domain is cleaved by furin, and the M1-GFP receptor travels to the plasma membrane. We plan to explore the factors that influence the plasma membrane expression of the M1 receptor using this fusion protein and TIRF.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目及 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 中心，不一定是研究者的机构。 我们计划使用由条件聚集域（CAD）与人M1受体N端融合组成的融合蛋白来研究中国仓鼠卵巢（CHO）细胞质膜上M1毒蕈碱受体表达的动力学和融合到 C 末端的绿色荧光蛋白 (GFP)。 在 CHO 细胞中瞬时表达后，融合蛋白 (GFP-M1-CAD) 被捕获在 ER 中，但在应用配体 (AP21998) 后可被诱导退出，该配体与 CAD 结合并导致融合蛋白聚集体在 ER 中解离。急诊室。 离开 ER 后，CAD 结构域被弗林蛋白酶切割，M1-GFP 受体进入质膜。 我们计划利用这种融合蛋白和 TIRF 来探索影响 M1 受体质膜表达的因素。