Photoswitchable cyan fluorescent protein as a FRET donor

Ekaterina A. Souslova, Dmitriy M. Chudakov

Research output: Contribution to journalArticlepeer-review

10 Citations (Scopus)

Abstract

Among a variety of fluorescent proteins available today, there is a lack of suitable markers with excitation/emission in the violet/blue part of visible spectrum. Recently, we reported on photoswitchable cyan fluorescent protein (PS-CFP), which represents monomeric high-contrasting photactivatable label for in vivo protein movement tracking. However, PS-CFP demands high intensity of light for the photoswitching. Therefore it can be employed as a common fluorescent tag at conventional light intensities, which cause negligible or zero photoactivation. High pH stability and unique positioning of excitation/emission peaks make it a worthy supplement to the existing palette of fluorescent proteins. Here we use PS-CFP fusion with a yellow fluorescent protein phiYFP to show that PS-CFP is a promising donor partner for the fluorescence resonance energy transfer (FRET). A remarkable phenomenon is that PS-CFP donor fluorescence turned to be essentially stable with and without FRET, while acceptor emission demonstrated record dynamic range of up to 7.8-fold. This makes the FRET pair presented a useful tool for the single color high throughput screenings. Here we also propose ways for further PS-CFP enhancing, aiming to develop bright cyan fluorescent protein with unique spectral characteristics.

Original languageEnglish
Pages (from-to)207-209
Number of pages3
JournalMicroscopy Research and Technique
Volume69
Issue number3
DOIs
Publication statusPublished - Mar 2006
Externally publishedYes

Keywords

  • Cyan fluorescent protein
  • Fluorescence resonance energy transfer
  • FRET
  • phiYFP
  • PS-CFP

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