A new bioluminescence-based tool for modulating target proteins in live cells

Sci Rep. 2019 Dec 3;9(1):18239. doi: 10.1038/s41598-019-54712-y.

Abstract

We have developed a new genetically encoded tool designed to generate reactive oxygen species (ROS) at target proteins in cultured cells; it is designed using firefly luciferase and photosensitiser protein KillerRed. Targeting this fusion protein, KillerFirefly, to F-actin in live cells and treatment with luciferin induced a characteristic structure, previously reported as a cofilin-actin rod, which is seen in patients with Alzheimer's disease. This structural change is considered to be elicited by the consistent generation of very low-level ROS by KillerFirefly in the vicinity of F-actin. Moreover, our results suggest the presence of an actin-regulating system, controlled by very low levels of endogenously generated ROS.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Actins / metabolism
  • Animals
  • Blotting, Western
  • CHO Cells / metabolism
  • Cells, Cultured
  • Cricetulus
  • Firefly Luciferin
  • Green Fluorescent Proteins
  • HEK293 Cells / metabolism
  • Humans
  • Luciferases
  • Luminescent Measurements / methods*
  • Photosensitizing Agents
  • Reactive Oxygen Species / metabolism
  • Recombinant Proteins

Substances

  • Actins
  • Photosensitizing Agents
  • Reactive Oxygen Species
  • Recombinant Proteins
  • killer red protein, Anthomedusae
  • Green Fluorescent Proteins
  • Firefly Luciferin
  • Luciferases