Far-red acclimating cyanobacterium as versatile source for bright fluorescent biomarkers.
Biochim Biophys Acta Mol Cell Res
; 1865(11 Pt A): 1649-1656, 2018 11.
Article
en En
| MEDLINE
| ID: mdl-30327206
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.
Palabras clave
Texto completo:
1
Colección:
01-internacional
Banco de datos:
MEDLINE
Asunto principal:
Proteínas Bacterianas
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Biomarcadores
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Cianobacterias
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Genes Reporteros
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Fluorescencia
Límite:
Animals
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Humans
Idioma:
En
Revista:
Biochim Biophys Acta Mol Cell Res
Año:
2018
Tipo del documento:
Article