Rational design of photoconvertible and biphotochromic fluorescent proteins for advanced microscopy applications.

Authors:
Dr. Virgile Adam, PhD
Dr. Virgile Adam, PhD
CNRS / Institute for Structural Biology (IBS)
Research scientist
single molecule fluorescence microscopy, fluorescent protein engineering, X-ray crystallography
Grenoble | France

Chem Biol 2011 Oct;18(10):1241-51

Laboratory of Photochemistry and Spectroscopy, Department of Chemistry, Katholieke Universiteit Leuven, 3001 Heverlee, Belgium.

Advanced fluorescence imaging, including subdiffraction microscopy, relies on fluorophores with controllable emission properties. Chief among these fluorophores are the photoactivatable fluorescent proteins capable of reversible on/off photoswitching or irreversible green-to-red photoconversion. IrisFP was recently reported as the first fluorescent protein combining these two types of phototransformations. The introduction of this protein resulted in new applications such as super-resolution pulse-chase imaging. However, the spectroscopic properties of IrisFP are far from being optimal and its tetrameric organization complicates its use as a fusion tag. Here, we demonstrate how four-state optical highlighting can be rationally introduced into photoconvertible fluorescent proteins and develop and characterize a new set of such enhanced optical highlighters derived from mEosFP and Dendra2. We present in particular NijiFP, a promising new fluorescent protein with photoconvertible and biphotochromic properties that make it ideal for advanced fluorescence-based imaging applications.

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Source
http://dx.doi.org/10.1016/j.chembiol.2011.08.007DOI Listing
October 2011
16 Reads
20 Citations
6.645 Impact Factor

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