Nature Biotechnology
21, 191 - 194 (2003)
Published online: 13 January 2003; | doi:10.1038/nbt778
There is a Corrigenda (April 2003) associated with this Technical Report.
Kindling fluorescent proteins for precise in vivo photolabelingDmitriy M. Chudakov1, 2, Vsevolod V. Belousov2, Andrey G. Zaraisky1, Vladimir V. Novoselov1, Dmitry B. Staroverov1, 2, Dmitry B. Zorov3, Sergey Lukyanov1, 2
& Konstantin A. Lukyanov1, 21
Shemiakin and Ovchinnikov Institute of Bioorganic Chemistry RAS, Miklukho-Maklaya 16/10, 117997 Moscow, Russia. 2
Evrogen, JSC, Miklukho-Maklaya 16/10, 117997 Moscow, Russia. 3
Belozersky Institute of Physical and Chemical Biology, Moscow State University, Khokhlov 6, 118899 Moscow, Russia.
Correspondence should be addressed to Konstantin A. Lukyanov kluk@ibch.ruPhotobleaching 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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