Photoswitchable cyan fluorescent protein for protein tracking

Dmitriy M. Chudakov, Vladislav V. Verkhusha, Dmitry B. Staroverov, Ekaterina A. Souslova, Sergey Lukyanov, Konstantin A. Lukyanov

Research output: Contribution to journalArticlepeer-review

288 Scopus citations

Abstract

In recent years diverse photolabeling techniques using green fluorescent protein (GFP)-like proteins have been reported1-7, including photoactivatable PA-GFP1, photoactivatable protein Kaede2, the DsRed 'greening' technique3 and kindling fluorescent proteins6,7. So far, only PA-GFP, which is monomeric and gives 100-fold fluorescence contrast, could be applied for protein tracking. Here we describe a dual-color monomeric protein, photoswitchable cyan fluorescent protein (PS-CFP). PS-CFP is capable of efficient photoconversion from cyan to green, changing both its excitation and emission spectra in response to 405-nm light irradiation. Complete photoactivation of PS-CFP results in a 1,500-fold increase in the green-to-cyan fluorescence ratio, making it the highest-contrast monomeric photoactivatable fluorescent protein described to date. We used PS-CFP as a photoswitchable tag to study trafficking of human dopamine transporter in living cells. At moderate excitation intensities, PS-CFP can be used as a pH-stable cyan label for protein tagging and fluorescence resonance energy transfer applications.

Original languageEnglish (US)
Pages (from-to)1435-1439
Number of pages5
JournalNature biotechnology
Volume22
Issue number11
DOIs
StatePublished - Nov 1 2004
Externally publishedYes

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology
  • Molecular Medicine
  • Biomedical Engineering

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