Cell Structure and Function
Online ISSN : 1347-3700
Print ISSN : 0386-7196
Live-cell Imaging with Genetically Encoded Protein Kinase Activity Reporters
Gembu MaryuHaruko MiuraYouichi UdaAkira T. KomatsubaraMichiyuki MatsudaKazuhiro Aoki
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2018 Volume 43 Issue 1 Pages 61-74

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Abstract

Protein kinases play pivotal roles in intracellular signal transduction, and dysregulation of kinases leads to pathological results such as malignant tumors. Kinase activity has hitherto been measured by biochemical methods such as in vitro phosphorylation assay and western blotting. However, these methods are less useful to explore spatial and temporal changes in kinase activity and its cell-to-cell variation. Recent advances in fluorescent proteins and live-cell imaging techniques enable us to visualize kinase activity in living cells with high spatial and temporal resolutions. Several genetically encoded kinase activity reporters, which are based on the modes of action of kinase activation and phosphorylation, are currently available. These reporters are classified into single-fluorophore kinase activity reporters and Förster (or fluorescence) resonance energy transfer (FRET)-based kinase activity reporters. Here, we introduce the principles of genetically encoded kinase activity reporters, and discuss the advantages and disadvantages of these reporters.

Key words: kinase, FRET, phosphorylation, KTR

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© 2018 The Author(s) CC-BY 4.0 (Submission before October 2016: Copyright © Japan Society for Cell Biology)
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