Manipulation of Actin Cytoskeleton by Intracellular-Targeted ROS Generation

Tetsuya Ishimoto*, Hisashi Mori

*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

4 Scopus citations

Abstract

A method to generate small amount of reactive oxygen species (ROSs) at intracellular targeted region has great potential to manipulate the function of particular proteins. The present protocol introduces a fusion protein that consisted of firefly luciferase (FLuc), photosensitizer protein KillerRed and F-actin-targeting peptide Lifeact (Lifeact-KillerFirefly) to generate ROSs in the vicinity of F-actin and found that morphological change in F-actin structure was induced by the fusion protein after luciferin treatment. This manipulating and imaging method is of use to analyze the role of the locally generated ROSs on the function of intracellular proteins.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages271-279
Number of pages9
DOIs
StatePublished - 2021

Publication series

NameMethods in Molecular Biology
Volume2274
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Actin polymerization
  • Bioluminescence resonance energy transfer
  • Cofilin-actin rod
  • Firefly luciferase (FLuc)
  • HEK293T
  • KillerRed
  • Photosensitizer protein
  • Reactive oxygen species

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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