TY - JOUR
T1 - Red fluorescent protein with reversibly photoswitchable absorbance for photochromic FRET
AU - Subach, Fedor V.
AU - Zhang, Lijuan
AU - Gadella, Theodorus W.J.
AU - Gurskaya, Nadya G.
AU - Lukyanov, Konstantin A.
AU - Verkhusha, Vladislav V.
N1 - Funding Information:
We thank A. Sorkin for providing plasmids, J. Zhang for assistance with flow cytometry, and L. Van Weeren for transfecting cells for FLIM. This work was supported by the NIH grant GM073913 (to V.V.V.), and in part by the Molecular and Cell Biology program of Russian Academy of Sciences, Rosobrazovanie grant P256, and President of the Russian Federation's grant MD-2780.2009.4 (to K.A.L.).
PY - 2010
Y1 - 2010
N2 - We have developed the first red fluorescent protein, named rsTagRFP, which possesses reversibly photoswitchable absorbance spectra. Illumination with blue and yellow light switches rsTagRFP into a red fluorescent state (ON state) or nonfluorescent state (OFF state), respectively. The ON and OFF states exhibit absorbance maxima at 567 and 440 nm, respectively. Due to the photoswitchable absorbance, rsTagRFP can be used as an acceptor for a photochromic Förster resonance energy transfer (pcFRET). The photochromic acceptor facilitates determination of a protein-protein interaction by providing an internal control for FRET. Using pcFRET with EYFP as a donor, we observed an interaction between epidermal growth factor receptor and growth factor receptor-binding protein 2 in live cells by detecting the modulation of both the fluorescence intensity and lifetime of the EYFP donor upon the ON-OFF photoswitching of the rsTagRFP acceptor.
AB - We have developed the first red fluorescent protein, named rsTagRFP, which possesses reversibly photoswitchable absorbance spectra. Illumination with blue and yellow light switches rsTagRFP into a red fluorescent state (ON state) or nonfluorescent state (OFF state), respectively. The ON and OFF states exhibit absorbance maxima at 567 and 440 nm, respectively. Due to the photoswitchable absorbance, rsTagRFP can be used as an acceptor for a photochromic Förster resonance energy transfer (pcFRET). The photochromic acceptor facilitates determination of a protein-protein interaction by providing an internal control for FRET. Using pcFRET with EYFP as a donor, we observed an interaction between epidermal growth factor receptor and growth factor receptor-binding protein 2 in live cells by detecting the modulation of both the fluorescence intensity and lifetime of the EYFP donor upon the ON-OFF photoswitching of the rsTagRFP acceptor.
KW - CHEMBIO
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U2 - 10.1016/j.chembiol.2010.05.022
DO - 10.1016/j.chembiol.2010.05.022
M3 - Article
C2 - 20659687
AN - SCOPUS:77955452640
SN - 1074-5521
VL - 17
SP - 745
EP - 755
JO - Chemistry and Biology
JF - Chemistry and Biology
IS - 7
ER -