Visualization of Compartmentalized Kinase Activity Dynamics Using Adaptable BimKARs. Issue 11 (19th November 2015)

• Record Type: Journal Article
• Authors: Depry, Charlene; Mehta, Sohum; Li, Ruojing; Zhang, Jin
• Is Part Of: Chemistry & biology. Volume 22:Issue 11(2015)
• Page Start: 1470
• Page End: 1479
• Publication Date: 2015-11-19
• Subjects: PKA -- AMPK -- FRET -- live-cell imaging -- biosensor -- compartmentalized signaling; Biochemistry -- Periodicals; 540
• DOI: 10.1016/j.chembiol.2015.10.004
• Languages: English
• ISSNs: 1074-5521

Abstract

Summary: 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. Graphical Abstract: Highlights: The versatile bimKAR design is easily adapted to monitor various kinase activities A membrane-targeted bimKAR improves the detection of local AMPK activity Membrane-localized AMPK activity undergoes opposing bidirectional regulation by PKA Abstract : Depry et al. demonstrate the versatility of FRET-based bimolecular kinase activity reporters (bimKARs) as tools for improving the visualization of signaling dynamics in living cells, and reveal that PKA acts simultaneously as both a positive and negative regulator of AMPK signaling at the plasma membrane.