Live-Cell Copper-Induced Fluorescence Quenching of the Flavin-Binding Fluorescent Protein CreiLOV.,ChemBioChem

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Live-Cell Copper-Induced Fluorescence Quenching of the Flavin-Binding Fluorescent Protein CreiLOV.
ChemBioChem ( IF 2.6 ) Pub Date : 2020-01-21 , DOI: 10.1002/cbic.201900669
Wenping Zou ₁ , Khoa Le ₁ , Melissa L Zastrow ₁

Affiliation

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.

中文翻译：

黄素结合荧光蛋白CreiLOV的活细胞铜诱导的荧光淬灭。

CreiLOV是一种黄素结合型荧光蛋白，衍生自包含光氧电压（LOV）感应域的蓝光感光蛋白家族。黄素结合型荧光蛋白代表了新的荧光报告基因和生物传感器的有前途的基础，它们可以解决公认的绿色荧光蛋白（GFP）家族的局限性。与黄素结合的荧光蛋白比GFP小，在较宽的pH范围内稳定，提供快速的生色团掺入，并且不依赖氧，因此可以应用于活的厌氧生物。在结合黄素的荧光蛋白中，CreiLOV具有高量子产率和出色的光物理性质，使其有望用于细胞应用。在这里，我们研究了CreiLOV作为基于强度和荧光寿命的金属传感器的适用性。

更新日期：2020-01-21

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