M3 muscarinic acetylcholine receptor facilitates the endocytosis of mu opioid receptor mediated by morphine independently of the formation of heteromeric complexes.
M3 muscarinic acetylcholine receptor facilitates the endocytosis of mu opioid receptor mediated by morphine independently of the formation of heteromeric complexes.
复制标题
M3 毒蕈碱乙酰胆碱受体促进吗啡介导的 mu 阿片受体的内吞作用，与异聚复合物的形成无关。
DOI：
10.1016/j.cellsig.2017.04.006
复制
发表时间：
2017
影响因子：
4.8
通讯作者：
Lopez-Gimenez JF
中科院分区：
文献类型：
--
作者：
Lopez-Gimenez JF
在许多实验模型中观察到吗啡诱导μ阿片样物质（MOP）受体内化的效率低下，这构成了G蛋白偶联受体（GPCR）功能选择性的一个范例。我们最近报道，通过在同一细胞中共表达的5-HT_2A-5-羟色胺受体激活Gαq/11蛋白，促进吗啡促进的MOP受体内吞。为了探索不同的Gαq/11偶联的GPCR是否会模拟这种效应，产生了永久表达MOP-YFP受体沿着以诱导方式表达的FLAG-M3-Cerulean受体的双稳定Flp-In T-REx HEK 293细胞系。这些细胞的荧光显微镜检查显示，这两种受体的共分布，主要是在质膜区室化。同时刺激卡巴胆碱和吗啡促进MOP受体的内化，脱敏和下调，这种促进作用不依赖于PKC激活。免疫共沉淀实验表明，FLAG-M3-Cerulean/MOP-YFP受体以时间依赖性方式相互作用形成异聚复合物，即在FLAG-M3-Cerulean诱导表达96小时后检测到最强的相互作用。在这些实验条件下，用卡巴胆碱加吗啡处理细胞导致两种受体在分离的内吞囊泡内的内化，如通过共聚焦显微镜观察到的。在激动剂刺激后，对FLAG-M3-Cerulean和MOP-YFP受体观察到的这种运输分离表明这种蛋白质-蛋白质相互作用呈现时间和动态特性。此外，由FLAG-M3-Cerulean受体促进的MOP-YFP受体内化独立于异聚复合物的构成。
Morphine inefficiency to induce the internalization of mu opioid (MOP) receptors observed in numerous experimental models constitutes a paradigm of G-protein coupled receptor (GPCR) functional selectivity. We recently described that activation of Gαq/11proteins through 5-HT2Aserotonin receptors co-expressed in the same cells facilitates MOP receptor endocytosis promoted by morphine. In order to explore whether a different Gαq/11coupled GPCR would emulate this effect, a double stable Flp-In T-REx HEK293 cell line permanently expressing MOP-YFP receptors along with FLAG-M3-Cerulean receptors expressed in an inducible manner was generated. Fluorescence microscopy examination of these cells revealed a co-distribution of both receptors mainly compartmentalized in plasma membrane. Concurrent stimulation with carbachol and morphine promoted MOP receptor internalization, desensitization and down-regulation and this facilitation was not dependent on PKC activation. Co-immunoprecipitation experiments demonstrated that FLAG-M3-Cerulean/MOP-YFP receptors interact forming heteromeric complexes in a time depending manner, i.e. the strongest interaction was detected after 96 h of FLAG-M3-Cerulean induced expression. Under these experimental conditions, treatment of cells with carbachol plus morphine resulted in the internalization of both receptors within separated endocytic vesicles as visualized by confocal microscopy. This trafficking segregation observed for FLAG-M3-Cerulean and MOP-YFP receptors upon agonist stimulation suggests that this protein-protein interaction presents temporal and dynamic properties. Moreover, MOP-YFP receptor internalization facilitated by FLAG-M3-Cerulean receptors is independent of the constitution of heteromeric complexes.
登录
查看更多内容
检查 mu 阿片受体内吞作用在长期使用阿片类药物的有益和副作用中的作用：来自 mu-阿片类药理学新概念研讨会。
影响因子：
4.2
作者：
Whistler,JenniferL
通讯作者：
Whistler,JenniferL
四种人毒蕈碱乙酰胆碱受体的独特一级结构、配体结合特性和组织特异性表达。
影响因子：
11.4
作者：
Peralta Eg;A. Ashkenazi;J. Winslow;Douglas H. Smith;J. Ramachandran;D. Capon
通讯作者：
D. Capon
影响因子：
4.8
作者：
Alvarez-Curto, Elisa;Ward, Richard J.;Milligan, Graeme
通讯作者：
Milligan, Graeme
DOI：
10.1007/978-1-61779-126-0_2
发表时间：
2011-01-01
期刊：
RECEPTOR SIGNAL TRANSDUCTION PROTOCOLS, THIRD EDITION
影响因子：
--
作者：
Ward, Richard J.;Alvarez-Curto, Elisa;Milligan, Graeme
通讯作者：
Milligan, Graeme
DOI：
10.1073/pnas.95.12.7157
发表时间：
1998-06-09
影响因子：
11.1
作者：
Zhang, J;Ferguson, SSG;Caron, MG
通讯作者：
Caron, MG