title: Manipulation of endogenous kinase activity in living cells using photoswitchable inhibitory peptides.
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.
authors.affiliation: Department of Pharmacology and ‡Department of Cell Biology and Physiology, University of North Carolina at Chapel Hill , Chapel Hill, North Carolina 27599, United States.
authors.forename: Jason J
authors.initials: JJ
authors.lastname: Yi
authors.forename: Hui
authors.initials: H
authors.lastname: Wang
authors.forename: Marco
authors.initials: M
authors.lastname: Vilela
authors.forename: Gaudenz
authors.initials: G
authors.lastname: Danuser
authors.forename: Klaus M
authors.initials: KM
authors.lastname: Hahn
mesh_terms: D000818:Animals; D018554:Avena / Q000235:genetics; D019556:COS Cells; D002462:Cell Membrane / Q000737:chemistry / Q000378:metabolism; D002522:Chlorocebus aethiops; D017868:Cyclic AMP-Dependent Protein Kinases / Q000737:chemistry / Q000378:metabolism; D008958:Models, Molecular; D009219:Myosin-Light-Chain Kinase / Q000737:chemistry / Q000378:metabolism; D062308:Optogenetics; D010455:Peptides / Q000737:chemistry / Q000235:genetics / Q000378:metabolism / Q000528:radiation effects; D018462:Photobiology; D056949:Phototropins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism / Q000528:radiation effects*; D047428:Protein Kinase Inhibitors / Q000737:chemistry* / Q000378:metabolism / Q000528:radiation effects*
chemical_list: D010455:Peptides; D056949:Phototropins; D047428:Protein Kinase Inhibitors; D017868:Cyclic AMP-Dependent Protein Kinases; D009219:Myosin-Light-Chain Kinase
keywords: CREB; MLCK; PKA; membrane dynamics; optogenetics; peptide caging