Premium
Photoswitchable cyan fluorescent protein as a FRET donor
Author(s) -
Souslova Ekaterina A.,
Chudakov Dmitriy M.
Publication year - 2006
Publication title -
microscopy research and technique
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.536
H-Index - 118
eISSN - 1097-0029
pISSN - 1059-910X
DOI - 10.1002/jemt.20278
Subject(s) - cyan , förster resonance energy transfer , fluorescence , yellow fluorescent protein , fluorescent protein , chemistry , photochemistry , green fluorescent protein , biophysics , optics , biology , biochemistry , physics , gene
Among a variety of fluorescent proteins available today, there is a lack of suitable markers with excitation/emission in the violet/blue part of visible spectrum. Recently, we reported on photoswitchable cyan fluorescent protein (PS‐CFP), which represents monomeric high‐contrasting photactivatable label for in vivo protein movement tracking. However, PS‐CFP demands high intensity of light for the photoswitching. Therefore it can be employed as a common fluorescent tag at conventional light intensities, which cause negligible or zero photoactivation. High pH stability and unique positioning of excitation/emission peaks make it a worthy supplement to the existing palette of fluorescent proteins. Here we use PS‐CFP fusion with a yellow fluorescent protein phiYFP to show that PS‐CFP is a promising donor partner for the fluorescence resonance energy transfer (FRET). A remarkable phenomenon is that PS‐CFP donor fluorescence turned to be essentially stable with and without FRET, while acceptor emission demonstrated record dynamic range of up to 7.8‐fold. This makes the FRET pair presented a useful tool for the single color high throughput screenings. Here we also propose ways for further PS‐CFP enhancing, aiming to develop bright cyan fluorescent protein with unique spectral characteristics. Microsc. Res. Tech. 69:207–209, 2006. © 2006 Wiley‐Liss, Inc.