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 structureguided, 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.

Far-red acclimating cyanobacterium as versatile source for bright fluorescent biomarkers / Ding, Wen-Long; Hou, Ya-Nan; Tan, Zi-Zhu; Jiang, Su-Ping; Miao, Dan; Losi, Aba; Gartner, Wolfgang; Scheer, Hugo; Zhao, Kai-Hong. - In: BIOCHIMICA ET BIOPHYSICA ACTA-MOLECULAR CELL RESEARCH. - ISSN 0167-4889. - 1865:11(2018), pp. 1649-1656. [10.1016/j.bbamcr.2018.08.015]

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

Losi, Aba
Membro del Collaboration Group
;
GARTNER, WOLFGANG;
2018-01-01

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 structureguided, 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.
2018
Far-red acclimating cyanobacterium as versatile source for bright fluorescent biomarkers / Ding, Wen-Long; Hou, Ya-Nan; Tan, Zi-Zhu; Jiang, Su-Ping; Miao, Dan; Losi, Aba; Gartner, Wolfgang; Scheer, Hugo; Zhao, Kai-Hong. - In: BIOCHIMICA ET BIOPHYSICA ACTA-MOLECULAR CELL RESEARCH. - ISSN 0167-4889. - 1865:11(2018), pp. 1649-1656. [10.1016/j.bbamcr.2018.08.015]
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11381/2849361
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