Photoswitchable cyan fluorescent protein for protein tracking

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

Research output: Contribution to journalArticle

277 Citations (Scopus)

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 2004
Externally publishedYes

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Proteins
Green Fluorescent Proteins
Fluorescence
Human Trafficking
Dopamine Plasma Membrane Transport Proteins
Fluorescence Resonance Energy Transfer
Labels
Cells
Cyan Fluorescent Protein
Irradiation
Color
Light

ASJC Scopus subject areas

  • Microbiology

Cite this

Chudakov, D. M., Verkhusha, V., Staroverov, D. B., Souslova, E. A., Lukyanov, S., & Lukyanov, K. A. (2004). Photoswitchable cyan fluorescent protein for protein tracking. Nature Biotechnology, 22(11), 1435-1439. https://doi.org/10.1038/nbt1025

Photoswitchable cyan fluorescent protein for protein tracking. / Chudakov, Dmitriy M.; Verkhusha, Vladislav; Staroverov, Dmitry B.; Souslova, Ekaterina A.; Lukyanov, Sergey; Lukyanov, Konstantin A.

In: Nature Biotechnology, Vol. 22, No. 11, 11.2004, p. 1435-1439.

Research output: Contribution to journalArticle

Chudakov, DM, Verkhusha, V, Staroverov, DB, Souslova, EA, Lukyanov, S & Lukyanov, KA 2004, 'Photoswitchable cyan fluorescent protein for protein tracking', Nature Biotechnology, vol. 22, no. 11, pp. 1435-1439. https://doi.org/10.1038/nbt1025
Chudakov DM, Verkhusha V, Staroverov DB, Souslova EA, Lukyanov S, Lukyanov KA. Photoswitchable cyan fluorescent protein for protein tracking. Nature Biotechnology. 2004 Nov;22(11):1435-1439. https://doi.org/10.1038/nbt1025
Chudakov, Dmitriy M. ; Verkhusha, Vladislav ; Staroverov, Dmitry B. ; Souslova, Ekaterina A. ; Lukyanov, Sergey ; Lukyanov, Konstantin A. / Photoswitchable cyan fluorescent protein for protein tracking. In: Nature Biotechnology. 2004 ; Vol. 22, No. 11. pp. 1435-1439.
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