Manipulation of endogenous kinase activity in living cells using photoswitchable inhibitory peptides

Jason J. Yi, Hui Wang, Marco Vilela, Gaudenz Danuser, Klaus M. Hahn

40 Scopus citations

Abstract

Optogenetic control of endogenous signaling can be an important tool for probing cell behavior. Using the photoresponse of the LOV2 domain of Avena sativa phototropin 1, we developed analogues of kinase inhibitors whose activity is light dependent. Inhibitory peptides were appended to the Jα helix, where they potently inhibited kinases in the light but were sterically blocked from kinase interaction in the dark. Photoactivatable inhibitors for cyclic-AMP dependent kinase (PKA) and myosin light chain kinase (MLCK) are described, together with studies that shed light on proper positioning of the peptides in the LOV domain. These inhibitors altered endogenous signaling in living cells and produced light-dependent changes in cell morphodynamics. (Figure Presented).

Original languageEnglish (US)
Pages (from-to)788-795
Number of pages8
JournalACS Synthetic Biology
Volume3
Issue number11
DOIs
Publication statusPublished - Nov 21 2014

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