Visualization of Compartmentalized Kinase Activity Dynamics Using Adaptable BimKARs.
Visualization of Compartmentalized Kinase Activity Dynamics Using Adaptable BimKARs.
复制标题
DOI：
10.1016/j.chembiol.2015.10.004
复制
发表时间：
2015-11-19
影响因子：
--
通讯作者：
Zhang J
中科院分区：
文献类型：
--
作者：
Depry C;Mehta S;Li R;Zhang J
在活细胞中监测激酶活性动力学的能力极大地有助于研究信号事件如何时空调节。在这里，我们报告的适应性双分子激酶活性报告（bimKARs）作为分子工具，以提高实时可视化的激酶活性。我们证明了bimKAR设计是真正通用的，可用于监测各种激酶，包括JNK，ERK和AMPK。此外，bimKAR可以具有比它们的单分子对应物显著增强的动态范围，允许阐明先前不可检测的激酶活性动力学。使用这些新设计的bimKAR，我们研究了质膜中PKA对AMPK的调节，并证明了刺激和未刺激（即，基础）PKA活性分别增强和抑制AMPK活性。
The ability to monitor kinase activity dynamics in live cells greatly aids the study of how signaling events are spatiotemporally regulated. Here, we report on the adaptability of bimolecular Kinase Activity Reporters (bimKARs) as molecular tools to enhance the real-time visualization of kinase activity. We demonstrate that the bimKAR design is truly versatile and can be used to monitor a variety of kinases, including JNK, ERK, and AMPK. Furthermore, bimKARs can have significantly enhanced dynamic ranges over their unimolecular counterparts, allowing the elucidation of previously undetectable kinase activity dynamics. Using these newly designed bimKARs, we investigate the regulation of AMPK by PKA in the plasma membrane and demonstrate that stimulated and unstimulated (i.e., basal) PKA activity enhance and suppress stimulated AMPK activity, respectively.