使用适应性双激酶活性报告子（BimKARs）可视化区室化激酶活性动力学
Visualization of Compartmentalized Kinase Activity Dynamics Using Adaptable BimKARs.
作者信息
Depry Charlene, Mehta Sohum, Li Ruojing, Zhang Jin
机构信息
Department of Pharmacology and Molecular Sciences, The Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
Department of Pharmacology, University of California, San Diego, 9500 Gilman Drive, BRF2 1120, La Jolla, CA 92093, USA.
出版信息
Chem Biol. 2015 Nov 19;22(11):1470-1479. doi: 10.1016/j.chembiol.2015.10.004. Epub 2015 Nov 5.
Abstract
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 protein kinase A (PKA) in the plasma membrane, and demonstrate that PKA can both negatively and positively regulate AMPK activity in the same cell.
摘要
在活细胞中监测激酶活性动态变化的能力极大地有助于研究信号事件是如何在时空上受到调控的。在此，我们报告双分子激酶活性报告分子（bimKARs）作为分子工具在增强激酶活性实时可视化方面的适应性。我们证明bimKAR设计具有真正的通用性，可用于监测多种激酶，包括JNK、ERK和AMPK。此外，与单分子对应物相比，bimKARs的动态范围可显著增强，从而能够阐明以前无法检测到的激酶活性动态变化。利用这些新设计的bimKARs，我们研究了蛋白激酶A（PKA）在质膜中对AMPK的调控，并证明PKA可在同一细胞中对AMPK活性进行负向和正向调控。
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