Prof. Konstantin Lukyanov, Inst. of Bioorganic Chemistry, Moscow RUSSIA

Unusual fluorescent proteins: design and applications

The study and applications of fluorescent proteins has gone a long way from a subject of highly specialized research to a widely known and indispensable tool for in vivo labeling, and from a single known member¾Green Fluorescent Protein from Aequorea victoria¾to hundreds of natural proteins of different colors with still poorly studied phylogeny and biological significance [1].

My talk will be focused on fluorescent proteins with “unusual” spectral or biochemical characteristics. In particular, we developed KillerRed – phototoxic fluorescent protein capable of light-induced production of reactive oxygen species (ROS) [2]. KillerRed allows light-induced ROS production to be strictly localized to different compartments and subcompartments within a cell. It was shown that KillerRed-mediated oxidative stress at different locations results in clearly different molecular events and cell responses (e.g., triggering apoptosis for mitochondria-localized KillerRed, or DNA damage and temporal blockage of cell division for chromatin-localized KillerRed) [2, 3].

Recently, we designed CFP variant with an anionic tryptophan-based chromophore [4]. In contrast to green and red fluorescent proteins usually containing anionic tyrosine-based chromophores, no charged states of the CFP chromophore have been described before. We believe that switching between protonated and deprotonated tryptophan in chromophores of fluorescent proteins represents a new unexplored strategy to control their spectral properties.

1. Chudakov D.M., Matz M.V., Lukyanov S., Lukyanov K.A. Fluorescent proteins and their applications in imaging living cells and tissues. Physiol. Rev. 2010, 90, 1103-1163.
2. Bulina M.E., Chudakov D.M., Britanova O.V., Yanushevich Y.G., Staroverov D.B., Chepurnykh T.V., Merzlyak E.M., Shkrob M.A., Lukyanov S., Lukyanov K.A. A genetically encoded photosensitizer. Nature Biotechnol. 2006, 24, 95-99.
3. Serebrovskaya EO, Gorodnicheva TV, Ermakova GV, Solovieva EA, Sharonov GV, Zagaynova EV, Chudakov DM, Lukyanov S, Zaraisky AG, Lukyanov KA. Light-induced blockage of cell division with a chromatin-targeted phototoxic fluorescent protein. Biochem. J. 2011, 435, 65-71.

4. Sarkisyan KS, Yampolsky IV, Solntsev KM, Lukyanov SA, Lukyanov KA, Mishin AS. Tryptophan-based chromophore in fluorescent proteins can be anionic. Sci Rep. 2012, 2, 608.

For more information contact Dr. Kyril Solntsev (404-385-7131).