Rational Design of Photoconvertible and Biphotochromic Fluorescent Proteins for Advanced Microscopy Applications

Virgile Adam (1, 2) , Benjamien Moeyaert (2) , Charlotte c. David (2) , Hideaki Mizuno (2) , Mickaël Lelimousin (3) , Peter Dedecker (2) , Ryoko Ando , Atsushi Miyawaki , Jan Michiels (2) , Yves Engelborghs (2) , Johan Hofkens (2)

1 IBS - UMR 5075 - Institut de biologie structurale
2 KU Leuven - Catholic University of Leuven = Katholieke Universiteit Leuven
3 University of Oxford

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.

Domaines: Biophysique

Virgile Adam, Benjamien Moeyaert, Charlotte c. David, Hideaki Mizuno, Mickaël Lelimousin, et al.. Rational Design of Photoconvertible and Biphotochromic Fluorescent Proteins for Advanced Microscopy Applications. Chemistry and Biology, 2011, 18 (10), pp.1241-1251. ⟨10.1016/j.chembiol.2011.08.007⟩. ⟨hal-02389699⟩