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.
Gurskaya
Nadya G
NG
Institute of Bioorganic Chemistry, Russian Academy of Sciences, Miklukho-Maklaya 16/10, Moscow 117997, Russia.
Verkhusha
Vladislav V
VV
Shcheglov
Alexander S
AS
Staroverov
Dmitry B
DB
Chepurnykh
Tatyana V
TV
Fradkov
Arkady F
AF
Lukyanov
Sergey
S
Lukyanov
Konstantin A
KA
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