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Live‐Cell Copper‐Induced Fluorescence Quenching of the Flavin‐Binding Fluorescent Protein CreiLOV
Author(s) -
Zou Wenping,
Le Khoa,
Zastrow Melissa L.
Publication year - 2020
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.201900669
Subject(s) - flavin group , fluorescence , fluorescence in the life sciences , green fluorescent protein , chromophore , photochemistry , chemistry , quenching (fluorescence) , copper protein , bimolecular fluorescence complementation , cuvette , biophysics , förster resonance energy transfer , copper , biochemistry , biology , enzyme , physics , organic chemistry , quantum mechanics , gene
CreiLOV is a flavin‐binding fluorescent protein derived from the blue‐light photoreceptor protein family that contains light‐oxygen‐voltage (LOV) sensing domains. Flavin‐binding fluorescent proteins represent a promising foundation for new fluorescent reporters and biosensors that can address limitations of the well‐established green fluorescent protein (GFP) family. Flavin‐binding fluorescent proteins are smaller than GFPs, are stable over a wider pH range, offer rapid chromophore incorporation, and are oxygen‐independent so can be applied to live anaerobic organisms. Among the flavin‐binding fluorescent proteins, CreiLOV has a high quantum yield and excellent photophysical properties, making it promising for cellular applications. Here, we investigated the suitability of CreiLOV as an intensity‐ and fluorescence‐lifetime‐based metal sensor. CreiLOV selectively binds copper(II) over other biologically relevant metals with low‐micromolar affinity, resulting in fluorescence quenching and a decrease in the fluorescence lifetime that can be observed in cuvettes and live bacterial cells.