Visualization of Compartmentalized Kinase Activity Dynamics Using Adaptable BimKARs

Charlene Depry, Sohum Mehta, Ruojing Li, Jin Zhang

Research output: Contribution to journalArticle

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.

Original languageEnglish (US)
Pages (from-to)1470-1480
Number of pages11
JournalChemistry and Biology
Volume22
Issue number11
DOIs
StatePublished - Nov 19 2015

Keywords

  • AMPK
  • biosensor
  • compartmentalized signaling
  • FRET
  • live-cell imaging
  • PKA

ASJC Scopus subject areas

  • Biochemistry
  • Drug Discovery
  • Molecular Biology
  • Clinical Biochemistry
  • Molecular Medicine
  • Pharmacology

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