title: Optical control of cell signaling by single-chain photoswitchable kinases.
abstract: Protein kinases transduce signals to regulate a wide array of cellular functions in eukaryotes. A generalizable method for optical control of kinases would enable fine spatiotemporal interrogation or manipulation of these various functions. We report the design and application of single-chain cofactor-free kinases with photoswitchable activity. We engineered a dimeric protein, pdDronpa, that dissociates in cyan light and reassociates in violet light. Attaching two pdDronpa domains at rationally selected locations in the kinase domain, we created the photoswitchable kinases psRaf1, psMEK1, psMEK2, and psCDK5. Using these photoswitchable kinases, we established an all-optical cell-based assay for screening inhibitors, uncovered a direct and rapid inhibitory feedback loop from ERK to MEK1, and mediated developmental changes and synaptic vesicle transport in vivo using light.
authors.affiliation: Department of Bioengineering, Stanford University, Stanford, CA, USA.
authors.forename: Xin X
authors.identifier: 0000-0001-6681-1568
authors.initials: XX
authors.lastname: Zhou
authors.affiliation: Department of Bioengineering, Stanford University, Stanford, CA, USA.
authors.forename: Linlin Z
authors.initials: LZ
authors.lastname: Fan
authors.affiliation: Howard Hughes Medical Institute and Department of Biology, Stanford University, Stanford, CA, USA.
authors.forename: Pengpeng
authors.initials: P
authors.lastname: Li
authors.affiliation: Howard Hughes Medical Institute and Department of Biology, Stanford University, Stanford, CA, USA.
authors.forename: Kang
authors.initials: K
authors.lastname: Shen
authors.affiliation: Department of Bioengineering, Stanford University, Stanford, CA, USA. mzlin@stanford.edu.|Department of Neurobiology, Stanford University, Stanford, CA, USA.
authors.forename: Michael Z
authors.identifier: 0000-0002-0492-1961
authors.initials: MZ
authors.lastname: Lin
mesh_terms: D000818:Animals; D017173:Caenorhabditis elegans; D045744:Cell Line, Tumor; D004353:Drug Evaluation, Preclinical; D048049:Extracellular Signal-Regulated MAP Kinases / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000235:genetics / Q000528:radiation effects*; D006801:Humans; D008027:Light; D048369:MAP Kinase Kinase 1 / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000235:genetics / Q000528:radiation effects*; D009154:Mutation; D000072417:Protein Domains; D015202:Protein Engineering; D047428:Protein Kinase Inhibitors / Q000737:chemistry / Q000494:pharmacology*; D055503:Protein Multimerization; D015398:Signal Transduction
chemical_list: D047428:Protein Kinase Inhibitors; D048049:Extracellular Signal-Regulated MAP Kinases; D048369:MAP Kinase Kinase 1