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.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 我们计划使用由条件聚集结构域（CAD）组成的融合蛋白（CAD）融合到人M1受体和绿色含量蛋白（gfp）fused的N-末端，研究中国仓鼠卵巢（CHO）细胞质膜表达的动力学。 在CHO细胞中瞬时表达后，将融合蛋白（GFP-M1-CAD）捕获在ER中，但可以在使用配体（AP21998）后诱导退出，该配体（AP21998）与CAD结合并导致ER中融合蛋白聚集体的解离。 退出ER后，CAD结构域被Furin裂解，M1-GFP受体向质膜传播。 我们计划探索使用该融合蛋白和TIRF影响M1受体质膜表达的因素。