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Biochim Biophys Acta Mol Cell Res. 2018 Nov;1865(11 Pt A):1649-1656. doi: 10.1016/j.bbamcr.2018.08.015. Epub 2018 Aug 25.

Far-red acclimating cyanobacterium as versatile source for bright fluorescent biomarkers.

Author information

1
State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, PR China.
2
Dept. of Mathematical, Physical and Computer Sciences, University of Parma, Parma, Italy.
3
Institute for Analytical Chemistry, University of Leipzig, Germany.
4
Department Biologie I, Universität München, Menzinger Str. 67, D-80638 München, Germany.
5
State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, PR China. Electronic address: khzhao@163.com.

Abstract

Far-red and near-infrared emitting chromophores extend applications of fluorescent proteins to regions of maximal transmission of most tissues, but present considerable engineering challenges. Far-red adapting cyanobacteria generate a novel set of biliproteins. One of them, ApcF2, from a thermophilic cyanobacterium was subjected to structure-guided, site-directed random and specific mutagenesis, and was screened for bright far-red emission. We report the generation of chromoproteins, termed BDFPs, that are small, bind auto-catalytically the ubiquitous biliverdin as chromophore, express well, and retain their fluorescence in mammalian cells and in the nematode, C. elegans. They are, moreover, photostable and tolerate high temperature, low pH and chemical denaturation. Homo-bichromophoric tandems of these proteins improve labeling, while hetero-bichromophoric systems with large Stokes shifts are suitable for applications like FRET, multi-channel or super-resolution microscopy. The BDFPs compare favorably to other biliproteins and provide a novel, extremely versatile labeling tool-box.

KEYWORDS:

Biliprotein; Far-red spectral range; Fluorescence biomarker; Protein engineering; Tissue penetration

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