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.
Keywords:
CREB; MLCK; PKA; membrane dynamics; optogenetics; peptide caging.
Publication types
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Letter
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Avena / genetics
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COS Cells
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Cell Membrane / chemistry
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Cell Membrane / metabolism
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Chlorocebus aethiops
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Cyclic AMP-Dependent Protein Kinases / chemistry
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Cyclic AMP-Dependent Protein Kinases / metabolism
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Models, Molecular
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Myosin-Light-Chain Kinase / chemistry
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Myosin-Light-Chain Kinase / metabolism
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Optogenetics
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Peptides / chemistry
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Peptides / genetics
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Peptides / metabolism
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Peptides / radiation effects
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Photobiology
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Phototropins / chemistry*
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Phototropins / genetics
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Phototropins / metabolism
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Phototropins / radiation effects*
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Protein Kinase Inhibitors / chemistry*
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Protein Kinase Inhibitors / metabolism
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Protein Kinase Inhibitors / radiation effects*
Substances
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Peptides
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Phototropins
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Protein Kinase Inhibitors
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Cyclic AMP-Dependent Protein Kinases
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Myosin-Light-Chain Kinase