Engineering of a monomeric green-to-red photoactivatable fluorescent protein induced by blue light.
Green fluorescent protein (GFP) and GFP-like proteins represent invaluable genetically encoded fluorescent probes. In the last few years a new class of photoactivatable fluorescent proteins (PAFPs) capable of pronounced light-induced spectral changes have been developed. Except for tetrameric KFP1 (ref. 4), all known PAFPs, including PA-GFP, Kaede, EosFP, PS-CFP, Dronpa, PA-mRFP1 and KikGR require light in the UV-violet spectral region for activation through one-photon excitation--such light can be phototoxic to some biological systems. 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 degrees 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.
Institute of Bioorganic Chemistry, Russian Academy of Sciences, Miklukho-Maklaya 16/10, Moscow 117997, Russia.
Nadya G
NG
Gurskaya
Vladislav V
VV
Verkhusha
Alexander S
AS
Shcheglov
Dmitry B
DB
Staroverov
Tatyana V
TV
Chepurnykh
Arkady F
AF
Fradkov
Sergey
S
Lukyanov
Konstantin A
KA
Lukyanov
D005456:Fluorescent Dyes; D008027:Light; D008164:Luminescent Proteins / Q000032:analysis* / Q000737:chemistry / Q000235:genetics* / Q000528:radiation effects; D036641:Microscopy, Fluorescence, Multiphoton / Q000379:methods*; D010777:Photochemistry / Q000379:methods*; D015202:Protein Engineering / Q000379:methods*
D005456:Fluorescent Dyes; D008164:Luminescent Proteins