Local Generation and Imaging of Hydrogen Peroxide in Living Cells

Yulia A. Bogdanova, Carsten Schultz, Vsevolod V. Belousov

Research output: Contribution to journalArticle

8 Scopus citations

Abstract

Described here is a localized H2 O2 generation-detection system consisting of a yeast D-amino acid oxidase (DAAO) and two spectrally distinct variants of biosensor, HyPer2 and HyPerRed based on circularly permutated yellow and red fluorescent proteins, respectively, which enables spatiotemporal production and examination of the intracellular H2 O2 dynamics. The protocol describes using this system in a simple cell culture model. We provide detailed instructions on imaging of H2 O2 generated by the activated DAAO. The system can be easily optimized for various combinations of cell types, conditions and DAAO/sensor subcellular localizations.

Original languageEnglish (US)
Pages (from-to)117-127
Number of pages11
JournalCurrent protocols in chemical biology
Volume9
Issue number2
DOIs
StatePublished - Jun 19 2017
Externally publishedYes

Keywords

  • biosensors
  • D-amino acid oxidase
  • hydrogen peroxide
  • HyPer
  • HyPer2
  • HyPerRed
  • metabolic engineering
  • reactive oxygen species

ASJC Scopus subject areas

  • Medicine(all)

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