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

Authors: Duanjun Cai, Miguel Alexandre Lopes Marques, Fernando Nogueira

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

Abstract: The firefly luciferase has been a unique marking tool used in various bioimaging techniques. Extensive color modulation is strongly required to meet special marking demands; however, intentional and accurate wavelength tuning has yet to be achieved. Here, we demonstrate that the color shift of the firefly chromophore (OxyLH2-1) by internal and external fields can be described as a unified Stark shift. Electrostatic microenvironmental effects on fluorescent spectroscopy are modeled in vacuo through effective electric fields by using time- dependent density functional theory. A complete visible fluorescence spectrum of firefly chromophore is depicted, which enables one to control the emission in a specific color. As an application, the widely observed pH-correlated color shift is proved to be associated with the local Stark field generated by the trace water−ions (vicinal hydronium and hydroxide ions) at active sites close to the OxyLH2-1.

DOI: 10.1021/jp405665v