TY - JOUR
T1 - Photoswitchable cyan fluorescent protein for protein tracking
AU - Chudakov, Dmitriy M.
AU - Verkhusha, Vladislav V.
AU - Staroverov, Dmitry B.
AU - Souslova, Ekaterina A.
AU - Lukyanov, Sergey
AU - Lukyanov, Konstantin A.
N1 - Funding Information:
We are grateful to Alexander Sorkin for cDNA of hDAT and valuable discussions. We thank Natalia E. Yelina for the critical reading of the manuscript. This work was supported by grants from National Institutes of Health/National Institute on Drug Abuse A DA014204 and National Institutes of Health/National Institute of General Medical Sciences GM070358 (to V.V.V.), Russian Academy of Sciences for the program ‘‘Molecular and Cell Biology’’ and EC FP-6 Integrated Project LSHG-CT-2003-503259.
PY - 2004/11
Y1 - 2004/11
N2 - In recent years diverse photolabeling techniques using green fluorescent protein (GFP)-like proteins have been reported1-7, including photoactivatable PA-GFP1, photoactivatable protein Kaede2, the DsRed 'greening' technique3 and kindling fluorescent proteins6,7. So far, only PA-GFP, which is monomeric and gives 100-fold fluorescence contrast, could be applied for protein tracking. Here we describe a dual-color monomeric protein, photoswitchable cyan fluorescent protein (PS-CFP). PS-CFP is capable of efficient photoconversion from cyan to green, changing both its excitation and emission spectra in response to 405-nm light irradiation. Complete photoactivation of PS-CFP results in a 1,500-fold increase in the green-to-cyan fluorescence ratio, making it the highest-contrast monomeric photoactivatable fluorescent protein described to date. We used PS-CFP as a photoswitchable tag to study trafficking of human dopamine transporter in living cells. At moderate excitation intensities, PS-CFP can be used as a pH-stable cyan label for protein tagging and fluorescence resonance energy transfer applications.
AB - In recent years diverse photolabeling techniques using green fluorescent protein (GFP)-like proteins have been reported1-7, including photoactivatable PA-GFP1, photoactivatable protein Kaede2, the DsRed 'greening' technique3 and kindling fluorescent proteins6,7. So far, only PA-GFP, which is monomeric and gives 100-fold fluorescence contrast, could be applied for protein tracking. Here we describe a dual-color monomeric protein, photoswitchable cyan fluorescent protein (PS-CFP). PS-CFP is capable of efficient photoconversion from cyan to green, changing both its excitation and emission spectra in response to 405-nm light irradiation. Complete photoactivation of PS-CFP results in a 1,500-fold increase in the green-to-cyan fluorescence ratio, making it the highest-contrast monomeric photoactivatable fluorescent protein described to date. We used PS-CFP as a photoswitchable tag to study trafficking of human dopamine transporter in living cells. At moderate excitation intensities, PS-CFP can be used as a pH-stable cyan label for protein tagging and fluorescence resonance energy transfer applications.
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U2 - 10.1038/nbt1025
DO - 10.1038/nbt1025
M3 - Article
C2 - 15502815
AN - SCOPUS:8344285764
SN - 1087-0156
VL - 22
SP - 1435
EP - 1439
JO - Nature biotechnology
JF - Nature biotechnology
IS - 11
ER -