TY - JOUR
T1 - Engineering of a monomeric green-to-red photoactivatable fluorescent protein induced by blue light
AU - Gurskaya, Nadya G.
AU - Verkhusha, Vladislav V.
AU - Shcheglov, Alexander S.
AU - Staroverov, Dmitry B.
AU - Chepurnykh, Tatyana V.
AU - Fradkov, Arkady F.
AU - Lukyanov, Sergey
AU - Lukyanov, Konstantin A.
N1 - Funding Information:
We thank Robert Goldman for providing us with the EGFP-vimentin vector and Alexey V. Feofanov for help in light intensity measurements. This work was supported by grants from the European Commission FP-6 Integrated Project LSHG-CT-2003-503259 (K.A.L.), the Russian Academy of Sciences for the program Molecular and Cell Biology (S.L.) and by grants GM070358 and DA019980 from the National Institutes of Health (V.V.V.).
PY - 2006/4
Y1 - 2006/4
N2 - 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.
AB - 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.
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U2 - 10.1038/nbt1191
DO - 10.1038/nbt1191
M3 - Article
C2 - 16550175
AN - SCOPUS:33645747837
SN - 1087-0156
VL - 24
SP - 461
EP - 465
JO - Nature biotechnology
JF - Nature biotechnology
IS - 4
ER -