Manipulation of Endogenous Kinase Activity in Living Cells Using Photoswitchable Inhibitory Peptides

作者：Jason J. Yi, Hui Wang, Marco Vilela, Gaudenz Danuser, Klaus M. Hahn · 发表于：ACS Synthetic Biology · 年份：2014 · DOI：10.1021/sb5001356 · 被引用次数：72 · 研究领域：Light effects on plants、Photochromic and Fluorescence Chemistry、Neurobiology and Insect Physiology Research

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.