| Literature DB >> 34050479 |
Tetsuya Ishimoto1, Hisashi Mori2.
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.Entities:
Keywords: Actin polymerization; Bioluminescence resonance energy transfer; Cofilin-actin rod; Firefly luciferase (FLuc); HEK293T; KillerRed; Photosensitizer protein; Reactive oxygen species
Year: 2021 PMID： 34050479 DOI： 10.1007/978-1-0716-1258-3_23
Source DB: PubMed Journal: Methods Mol Biol ISSN： 1064-3745