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Kindling fluorescent proteins for precise in vivo photolabeling

Nature Biotechnology volume 21, pages 191–194 (2003)Cite this article

A Corrigendum to this article was published on 01 April 2003

Abstract

Photobleaching of green fluorescent protein (GFP) is a widely used approach for tracking the movement of subcellular structures and intracellular proteins1,2,3. Although photobleaching is a powerful technique, it does not allow direct tracking of an object's movement and velocity within a living cell. Direct tracking becomes possible only with the introduction of a photoactivated fluorescent marker. A number of previous studies have reported optically induced changes in the emission spectra of fluorescent proteins4,5,6,7. However, the ideal photoactivated fluorescent marker should be a nonfluorescent tag capable of “switching on” (i.e., becoming fluorescent) in response to irradiation by light of a particular wavelength, intensity, and duration. In this report, we generated a mutant of Anemonia sulcata chromoprotein asCP8. The mutant protein is capable of unique irreversible photoconversion from the nonfluorescent to a stable bright-red fluorescent form (“kindling”). This “kindling fluorescent protein” (KFP1) can be used for precise in vivo photolabeling to track the movements of cells, organelles, and proteins. We used KFP1 for in vivo cell labeling in mRNA microinjection assays to monitor Xenopus laevis embryo development and to track mitochondrial movement in mammalian cells.

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Figure 1: KFP1 and asCP kindling.
Figure 2: Xenopus laevis embryo development imaged using KFP1.
Figure 3: Mitochondrial tracking within the eukaryotic cell using KFP1.

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Acknowledgements

The authors thank A.V. Feofanov (Shemiakin and Ovchinnikov Institute of Bioorganic Chemistry RAS) for valuable help in intracellular fluorescence kindling technique development. This work was supported by the Russian Foundation for Basic Research (grant 01-04-49037), the Russian Foundation for Support of Domestic Science grant to S.A.L., and by grants from HHMI (55000344), FIRCA, and CRDF (RB1-2406-MO-02) to A.G.Z.

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Authors and Affiliations

  1. Shemiakin and Ovchinnikov Institute of Bioorganic Chemistry RAS, Miklukho-Maklaya 16/10, Moscow, 117997, Russia

    Dmitriy M. Chudakov, Andrey G. Zaraisky, Vladimir V. Novoselov, Dmitry B. Staroverov, Sergey Lukyanov & Konstantin A. Lukyanov

  2. Evrogen, JSC, Miklukho-Maklaya 16/10, Moscow, 117997, Russia

    Dmitriy M. Chudakov, Vsevolod V. Belousov, Dmitry B. Staroverov, Sergey Lukyanov & Konstantin A. Lukyanov

  3. Belozersky Institute of Physical and Chemical Biology, Moscow State University, Khokhlov 6, Moscow, 118899, Russia

    Dmitry B. Zorov

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  1. Dmitriy M. Chudakov
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  2. Vsevolod V. Belousov
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Correspondence to Konstantin A. Lukyanov.

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Chudakov, D., Belousov, V., Zaraisky, A. et al. Kindling fluorescent proteins for precise in vivo photolabeling. Nat Biotechnol 21, 191–194 (2003). https://doi.org/10.1038/nbt778

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