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Full Color Modulation of Firefly Luciferase through Engineering with Unified Stark Effect

Authors: Duanjun Cai, M.A.L. Marques, and Fernando Nogueira

Ref.: J. Phys. Chem. B 117, 13725-13730 (2013)

Abstract: The firefly luciferin/luciferase system, a famous example of fluorescent proteins, has been successfully used in various bio-imaging techniques, as a reporter for ATP generation, gene expression, and as a biosensor for environmental pollutants. In all these diverse applications, some special marking demands require an extensive color modulation of the fluorescent properties of the protein. For this purpose, mechanisms resulting in or leading to, e.g., color shift, color tuning, and brightness enhancement have been actively studied. However, due to the ambiguity and complexity of the micro-environmental effects on the emitted light of the chromophore, really intentional and accurate wavelength modulation has yet to be achieved. In this work, we proposed a new concept of unified Stark effect and established the complete theory and method for full color modulation of firefly protein through charged residue mutagenesis technique. The unified Stark effect has covered the important internal and external influences by surrounding environmental electric interactions. This concept well explains various color shift phenomena in fluorescent proteins, including the clarification of the widely discussed pH-dependent color shift. Based on this concept, a complete full-color fluorescence map of firefly chromophore is successfully depicted, which provides clear guidance for the site-directed mutagenesis to approach intentional and accurate color tuning on fluorescent protein system. It is believed that this approach will be beneficial for bioluminescence researches and also the functionalization of organic optoelectronic devices.

Citations: 18 (Google scholar)

DOI: 10.1021/jp405665v

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Bibtex:

@article{Cai_2013,
	doi = {10.1021/jp405665v},
	url = {https://doi.org/10.1021%2Fjp405665v},
	year = 2013,
	month = {oct},
	publisher = {American Chemical Society ({ACS})},
	volume = {117},
	number = {44},
	pages = {13725--13730},
	author = {Duanjun Cai and Miguel A. L. Marques and Fernando Nogueira},
	title = {Full Color Modulation of Firefly Luciferase through Engineering with Unified Stark Effect},
	journal = {The Journal of Physical Chemistry B}
}