Heterogeneity of signal transduction at the subcellular level: microsphere-based focal EGF receptor activation and stimulation of Shc translocation.
Heterogeneity of signal transduction at the subcellular level: microsphere-based focal EGF receptor activation and stimulation of Shc translocation.
复制标题
亚细胞水平信号转导的异质性：基于微球的局灶 EGF 受体激活和 Shc 易位刺激。
作者：
Roland Brock;T. Jovin
表皮生长因子受体（EGFR，erbB 1）的激活和易位的Shc衔接蛋白激活受体进行了分析，在亚细胞水平的双标记免疫荧光和共聚焦激光扫描显微镜结合一个新的微球为基础的协议。在这里介绍的定量微球募集试验（QMRA）中，表皮生长因子包被的1 μ m直径的微球分布在表达受体的贴壁组织培养细胞的表面上。高分辨率共聚焦显微镜的受体和绿色荧光蛋白在中国仓鼠卵巢细胞中表达的融合结构表明，吞噬和内化的微球迅速发生在几分钟内，并在受体激活依赖性的方式。在人表皮样癌A431细胞中，受体活化和Shc易位持续超过20分钟的实验时间过程。然而，在亚细胞水平上，在5-8分钟观察到的受体活化和Shc易位的正相关性消失，表明两个事件的时间依赖性解耦和不同亚细胞位置的信号转导动力学的变化。
Epidermal growth factor receptor (EGFR, erbB1) activation and translocation of the Shc adaptor protein to activated receptors were analyzed at the subcellular level by dual-label immunofluorescence and confocal laser scanning microscopy in conjunction with a new microsphere-based protocol. In the Quantitative Microsphere Recruitment Assay (QMRA) introduced here, epidermal growth factor-coated 1 microm diameter microspheres were distributed over the surface of adherent tissue culture cells expressing the receptor. High-resolution confocal microscopy of a fusion construct of the receptor and the green fluorescent protein expressed in Chinese hamster ovary cells demonstrated that engulfment and internalization of the microspheres occurred rapidly within minutes, and in a receptor activation-dependent manner. In human epidermoid carcinoma A431 cells, receptor activation and Shc translocation persisted over the 20-minute time course of the experiments. However, at the subcellular level the positive correlation of receptor activation and Shc translocation observed at 5-8 minutes dissipated, indicating a time-dependent decoupling of the two events and variation in the kinetics of signal transduction for different subcellular locations.
通过与去污剂不溶性细胞骨架结合增强表皮生长因子受体酪氨酸激酶活性的证据：完整和羧基末端截短的受体的分析。
影响因子：
4.8
作者：
Gronowski,AM;Bertics,PJ
通讯作者：
Bertics,PJ
DOI：
10.1073/pnas.93.19.10078
发表时间：
1996-09-17
影响因子：
11.1
作者：
Huang, CYF;Ferrell, JE
通讯作者：
Ferrell, JE