Engineering of a monomeric green-to-red photoactivatable fluorescent protein induced by blue light

Nadya G. Gurskaya, Vladislav V. Verkhusha, Alexander S. Shcheglov, Dmitry B. Staroverov, Tatyana V. Chepurnykh, Arkady F. Fradkov, Sergey Lukyanov, Konstantin A. Lukyanov

Research output: Contribution to journalArticle

487 Scopus citations

Abstract

Green fluorescent protein (GFP) and GFP-like proteins represent invaluable genetically encoded fluorescent probes1,2. In the last few years a new class of photoactivatable fluorescent proteins (PAFPs) capable of pronounced light-induced spectral changes have been developed3. Except for tetrameric KFP1 (ref. 4), all known PAFPs, including PA-GFP5, Kaede6, EosFP7, PS-CFP8, Dronpa9, PA-mRFP110 and KikGR11 require light in the UV-violet spectral region for activation through one-photon excitation - such light can be phototoxic to some biological systems12. Here, we report a monomeric PAFP, Dendra, derived from octocoral Dendronephthya sp. and capable of 1,000- to 4,500-fold photoconversion from green to red fluorescent states in response to either visible blue or UV-violet light. Dendra represents the first PAFP, which is simultaneously monomeric, efficiently matures at 37°C, demonstrates high photostability of the activated state, and can be photoactivated by a common, marginally phototoxic, 488-nm laser line. We demonstrate the suitability of Dendra for protein labeling and tracking to quantitatively study dynamics of fibrillarin and vimentin in mammalian cells.

Original languageEnglish (US)
Pages (from-to)461-465
Number of pages5
JournalNature biotechnology
Volume24
Issue number4
DOIs
StatePublished - Apr 1 2006

ASJC Scopus subject areas

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

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    Gurskaya, N. G., Verkhusha, V. V., Shcheglov, A. S., Staroverov, D. B., Chepurnykh, T. V., Fradkov, A. F., Lukyanov, S., & Lukyanov, K. A. (2006). Engineering of a monomeric green-to-red photoactivatable fluorescent protein induced by blue light. Nature biotechnology, 24(4), 461-465. https://doi.org/10.1038/nbt1191