Ultrafast fluorescence depolarisation in green fluorescence protein tandem dimers as hydrophobic environment sensitive probes

Advances in ultra-fast photonics have enabled monitoring of biochemical interactions on a sub nano-second time scale. In addition, picosecond dynamics of intermolecular energy transfer in fluorescent proteins has been observed. Here, we present the development of a genetically encoded fluorescent se...

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Bibliographic Details
Published in:Physical chemistry chemical physics : PCCP 2023-07, Vol.25 (29), p.19532-19539
Main Authors: Sánchez-Pedreño Jiménez, Alejandro, Puhl, Henry L, Vogel, Steven S, Kim, Youngchan
Format: Article
Language:English
Subjects:
Amino acids
Anisotropy
Chemical compounds
Chemistry
Depolarization
Dimers
Energy transfer
Fluorescence
Genetic code
Glycerol
Hydrophobicity
Monitoring
Proteins
Sensors
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Summary:Advances in ultra-fast photonics have enabled monitoring of biochemical interactions on a sub nano-second time scale. In addition, picosecond dynamics of intermolecular energy transfer in fluorescent proteins has been observed. Here, we present the development of a genetically encoded fluorescent sensor that can detect changes in hydrophobicity by monitoring ultrafast fluorescence depolarisation. Our sensor is composed of a pair of dimeric enhanced green fluorescent proteins (dEGFPs) linked by a flexible amino-acid linker. We show dimerisation is perturbed by the addition of glycerol which interferes with the hydrophobic interaction of the two proteins. Time-resolved fluorescence anisotropy revealed a systematic attenuation of ultrafast fluorescence depolarisation when the sensor was exposed to increasing glycerol concentrations. This suggests that as hydrophobicity increases, dEGFP pairing decreases within a tandem dimer. Un-pairing of the protein fluorophores dramatically alters the rate of energy transfer between the proteins, resulting in an increase in the limiting anisotropy of the sensor. Increasing glycerol concentrations led to a systematic attenuation of ultrafast fluorescence depolarisation in dEGFP-TD.
ISSN:1463-9076
1463-9084
DOI:10.1039/d3cp01765f