Photoswitchable cyan fluorescent protein for protein tracking.
In recent years diverse photolabeling techniques using green fluorescent protein (GFP)-like proteins have been reported, including photoactivatable PA-GFP, photoactivatable protein Kaede, the DsRed 'greening' technique and kindling fluorescent proteins. 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.
Chudakov
Dmitriy M
DM
Institute of Bioorganic Chemistry RAS, Laboratory of Genes for Regeneration, Miklukho-Maklaya 16/10, Moscow 117997, Russia.
Verkhusha
Vladislav V
VV
Staroverov
Dmitry B
DB
Souslova
Ekaterina A
EA
Lukyanov
Sergey
S
Lukyanov
Konstantin A
KA
D000595:Amino Acid Sequence; D004926:Escherichia coli / Q000166:cytology / Q000378:metabolism*; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D049452:Green Fluorescent Proteins / Q000737:chemistry* / Q000528:radiation effects; D008856:Microscopy, Fluorescence / Q000379:methods*; D008969:Molecular Sequence Data; D010777:Photochemistry / Q000379:methods; D021381:Protein Transport / Q000502:physiology*; D011994:Recombinant Proteins / Q000737:chemistry / Q000528:radiation effects
C485182:Cyan Fluorescent Protein; D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins