Development and Characterization of Flavin-Binding Fluorescent Proteins, Part II: Advanced Characterization

Methods Mol Biol. 2023:2564:143-183. doi: 10.1007/978-1-0716-2667-2_7.

Abstract

Flavin-based fluorescent proteins (FbFPs), a class of small fluorescent proteins derived from light-oxygen-voltage (LOV) domains, bind ubiquitous endogenous flavins as chromophores. Due to their unique properties, they can be used as versatile in vivo reporter proteins under aerobic and anaerobic conditions. This chapter presents methodologies for in-depth characterization of the biochemical, spectroscopic, photophysical, and photochemical properties of FbFPs.

Keywords: Flavin; Flavin-binding fluorescent protein; Fluorescence brightness; Fluorescence lifetime; Fluorescence microscopy; Genetically encoded photosensitizer; LOV domain; Phototoxicity.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Dinitrocresols*
  • Flavins* / metabolism
  • Oxygen / metabolism
  • Proteins

Substances

  • Dinitrocresols
  • Flavins
  • Proteins
  • 4,6-dinitro-o-cresol
  • Oxygen