Developing a genetically encoded green fluorescent protein mutant for sensitive light-up fluorescent sensing and cellular imaging of Hg(II)

Jiang, Tao; Guo, Daiping; Wang, Qian; Wu, Xin; Li, Zhao; Zheng, Zhenhua; Yin, Boyuan; Lin, Xia; Tang, Jixian; Luo, Wenxin; Xia, Ningshao; Jiang, Yun‐Bao

doi:10.1016/j.aca.2015.03.026

Cited by 21 publications

(14 citation statements)

References 45 publications

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“…Moreover, combination of natural ligand binding proteins with fluorescent proteins allows large dynamic range . In consistent with this study, we similarly obtained a larger dynamic range compared with that of GFP mutant .…”

Section: Resultssupporting

confidence: 89%

“…As summarized in Table , MerR:EYFP chimera biosensor showed higher sensitivity when compared with four of the previously reported single fluorescent protein biosensors . Although the LOD values of GFP mutant biosensor and MerR:EYFP chimera biosensor were same, the highest detectable Hg 2+ concentration was more than 10‐fold higher for MerR:EYFP. Metal‐binding peptides have also been used to design Hg sensors, albeit with a lower sensitivity than MerR:EYFP chimera biosensor.…”

Section: Resultsmentioning

confidence: 79%

“…A similar fluorescence quenching‐based Hg 2+ biosensor was obtained by introducing cysteine residue at position 205 of GFP . The histidine to cysteine mutation at position 148 created a GFP variant with enhanced fluorescence response upon addition of Hg 2+ . Blocking of the chromophore conjugation by Hg 2+ provided a turn‐off infrared fluorescent protein probe .…”

Section: Introductionmentioning

confidence: 99%

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MerR‐fluorescent protein chimera biosensor for fast and sensitive detection of Hg²⁺ in drinking water

Özyurt

Üstükarcı

Evran

et al. 2019

Biotech and App Biochem

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Mercury ion (Hg2+) is a universal pollutant and its detection is crucial for public healthcare. In this study, we developed a novel fluorescent biosensor by construction of a protein fusion between the mercury‐sensing transcription factor MerR and enhanced yellow fluorescent protein (EYFP). Hg2+‐induced conformational change of MerR was transduced into fluorescence signal. Fluorescence intensity of the biosensor protein decreased with increasing concentrations of Hg2+ and a linear response was obtained in the range of 0.5–40 nM. The limit of detection was 0.5 nM, which was much lower than the maximum allowed level in water. The biosensor specificity was highly dependent on type and concentration of metal ion. The biosensor exhibited high specificity in a mixture of metal ions at 0.5 nM concentration. However, the interference effect was more pronounced at 40 nM concentration of metal ions. The measurement was completed in less than 1 Min with no need for sample preparation or preincubation steps. The biosensor achieved accurate and reliable detection in the spiked drinking water sample, as validated by the inductively coupled plasma optical emission spectrometry.

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Section: Resultssupporting

confidence: 89%

Section: Resultsmentioning

confidence: 79%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

MerR‐fluorescent protein chimera biosensor for fast and sensitive detection of Hg²⁺ in drinking water

Özyurt

Üstükarcı

Evran

et al. 2019

Biotech and App Biochem

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“…Upon binding to Hg 2+ , the fluorescence of FP could be either quenched 20, 21 or moderately enhanced. 22 In the present work, we devised a reaction-based Hg 2+ FP sensor, which was highly selective and displayed a large fluorescence enhancement in the presence of Hg 2+ .…”

mentioning

confidence: 99%

“…Several of these probes can be used to detect Hg 2+ in live cells. 12–15 Besides small-molecule probes, 12–17 protein-based, 18, 19 in particular fluorescent protein (FP)-based, 20–22 detection methods have been developed in recent years. The FP biosensors have the advantages in its genetic encodability and the flexibility of facile targeting into different sub-cellular compartments.…”

mentioning

confidence: 99%

Fluorescent Protein-Based Turn-On Probe through a General Protection–Deprotection Design Strategy

et al. 2017

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We demonstrated a general protection-deprotection strategy for the design of fluorescent protein biosensors through the construction of a turn-on Hg2+ sensor. A combination of fluorescent protein engineering and unnatural amino acid mutagenesis was used. Unlike previously reported fluorescent protein-based Hg2+ sensors that relied on the binding of Hg2+ to the sulfhydryl group of cysteine residues, a well-established chemical reaction, oxymercuration, was transformed into biological format and incorporated into our sensor design. This novel Hg2+ sensor displayed good sensitivity and selectivity both in vitro and in live bacterial cells. Over 60 folds change in fluorescence signal output was observed in the presence of 10 μM Hg2+, while such change was undetectable when nine other metal ions were tested. This new design strategy could expand the repertoire of fluorescent protein-based biosensors for the detection of small-molecule analytes.

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A highly selective fluorescent chemosensor for Cu²⁺ based on a diarylethene with a 2,1,3‐benzoxadiazole unit

Zhang

Liu

2018

J of Physical Organic Chem

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A novel photochromic diarylethene containing a 2,1,3-benzoxadiazole structure has been synthesized and its multicontrollable fluorescence properties have been systematically studied by the stimulation of light and metal ions. The compound exhibited favorable photochromism and fluorescent switching properties. Furthermore, it can serve as a fluorescent chemosensor for highly selective recognition of Cu 2+ in aqueous acetonitrile. When triggered by Cu 2+ , it displayed a remarkable blue shift from 594 to 524 nm (70 nm) with a significant fluorescence color change from yellow to bright green. As a result, a logic circuit has also been constructed on the basis of light and chemical stimulation.

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Developing a genetically encoded green fluorescent protein mutant for sensitive light-up fluorescent sensing and cellular imaging of Hg(II)

Cited by 21 publications

References 45 publications

MerR‐fluorescent protein chimera biosensor for fast and sensitive detection of Hg²⁺ in drinking water

MerR‐fluorescent protein chimera biosensor for fast and sensitive detection of Hg²⁺ in drinking water

Fluorescent Protein-Based Turn-On Probe through a General Protection–Deprotection Design Strategy

A highly selective fluorescent chemosensor for Cu²⁺ based on a diarylethene with a 2,1,3‐benzoxadiazole unit

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Developing a genetically encoded green fluorescent protein mutant for sensitive light-up fluorescent sensing and cellular imaging of Hg(II)

Cited by 21 publications

References 45 publications

MerR‐fluorescent protein chimera biosensor for fast and sensitive detection of Hg2+ in drinking water

MerR‐fluorescent protein chimera biosensor for fast and sensitive detection of Hg2+ in drinking water

Fluorescent Protein-Based Turn-On Probe through a General Protection–Deprotection Design Strategy

A highly selective fluorescent chemosensor for Cu2+ based on a diarylethene with a 2,1,3‐benzoxadiazole unit

Contact Info

Product

Resources

About

MerR‐fluorescent protein chimera biosensor for fast and sensitive detection of Hg²⁺ in drinking water

MerR‐fluorescent protein chimera biosensor for fast and sensitive detection of Hg²⁺ in drinking water

A highly selective fluorescent chemosensor for Cu²⁺ based on a diarylethene with a 2,1,3‐benzoxadiazole unit