Engineering of a Monomeric Green-To-Red Photoactivatable Fluorescent Protein Induced by Blue Light

Nat Biotechnol. 2006 Apr;24(4):461-5. doi: 10.1038/nbt1191. Epub 2006 Mar 19.

Abstract

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Fluorescent Dyes*
  • Light
  • Luminescent Proteins / analysis*
  • Luminescent Proteins / chemistry
  • Luminescent Proteins / genetics*
  • Luminescent Proteins / radiation effects
  • Microscopy, Fluorescence, Multiphoton / methods*
  • Photochemistry / methods*
  • Protein Engineering / methods*

Substances

  • Fluorescent Dyes
  • Luminescent Proteins