Red fluorescent protein with reversibly photoswitchable absorbance for photochromic FRET

Fedor V. Subach, Lijuan Zhang, Theodorus W.J. Gadella, Nadya G. Gurskaya, Konstantin A. Lukyanov, Vladislav V. Verkhusha

Research output: Contribution to journalArticle

100 Scopus citations

Abstract

We have developed the first red fluorescent protein, named rsTagRFP, which possesses reversibly photoswitchable absorbance spectra. Illumination with blue and yellow light switches rsTagRFP into a red fluorescent state (ON state) or nonfluorescent state (OFF state), respectively. The ON and OFF states exhibit absorbance maxima at 567 and 440 nm, respectively. Due to the photoswitchable absorbance, rsTagRFP can be used as an acceptor for a photochromic Förster resonance energy transfer (pcFRET). The photochromic acceptor facilitates determination of a protein-protein interaction by providing an internal control for FRET. Using pcFRET with EYFP as a donor, we observed an interaction between epidermal growth factor receptor and growth factor receptor-binding protein 2 in live cells by detecting the modulation of both the fluorescence intensity and lifetime of the EYFP donor upon the ON-OFF photoswitching of the rsTagRFP acceptor.

Original languageEnglish (US)
Pages (from-to)745-755
Number of pages11
JournalChemistry and Biology
Volume17
Issue number7
DOIs
Publication statusPublished - Aug 16 2010

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Keywords

  • CHEMBIO

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Medicine
  • Molecular Biology
  • Pharmacology
  • Drug Discovery
  • Clinical Biochemistry

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