In Situ Localization of Enzyme Activity in Live Cells by a Molecular Probe Releasing a Precipitating Fluorochrome

Liu, Hong‐Wen; Li, Ké; Hu, Xiaoxiao; Zhu, Longmin; Rong, Qiming; Liu, Yongchao; Zhang, Xiaobing; Hasserodt, Jens; Qu, Fengli; Tan, Weihong

doi:10.1002/anie.201705747

Cited by 181 publications

(127 citation statements)

References 43 publications

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“…Probe 87 was also used to visualise different ALP activity levels in Saos-2 and U-2OS osteosarcoma cells, thus enabling the role of ALP in the development of physiological and pathological processes to be determined. 217 Hu et al developed an AIE + ESIPT HPQ-based fluorescent probe 88 that was designed to allow for the enzymatic detection of b-galactosidase (b-Gal). This is an enzyme that catalyses the hydrolysis of oligosaccharides containing b-galactoside linkages.…”

Section: àmentioning

confidence: 99%

Excited-state intramolecular proton-transfer (ESIPT) based fluorescence sensors and imaging agents

et al. 2018

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Section: àmentioning

confidence: 99%

Excited-state intramolecular proton-transfer (ESIPT) based fluorescence sensors and imaging agents

et al. 2018

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“…Probe 36 was able to distinguish cells with different physiological profiles in heterogeneous tumor tissues. 81 In contrast to the sedimentary nature of probe 36 , near-infrared probe 37 was designed for dynamic imaging of ALP in live mice. 14 Probe 38 , which targets PTP, is a two-photon acyloxymethyl ketone probe conjugated to cell-penetrating peptides to facilitate organelle-specific detection at tissue depths of up to 100 μm.…”

Section: Hydrolases (Ec 3)mentioning

confidence: 99%

Enzyme-Activated Fluorogenic Probes for Live-Cell and in Vivo Imaging

2018

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Fluorogenic probes, small-molecule sensors that unmask brilliant fluorescence upon exposure to specific stimuli, are powerful tools for chemical biology. Those probes that respond to enzymatic catalysis illuminate the complex dynamics of biological processes at a level of spatiotemporal detail and sensitivity unmatched by other techniques. Here, we review recent advances in enzyme-activated fluorogenic probes for biological imaging. We organize our survey by enzyme classification, with emphasis on fluorophore masking strategies, modes of enzymatic activation, and the breadth of current and future applications. Key challenges such as probe selectivity and spectroscopic requirements are described alongside of therapeutic, diagnostic, and theranostic opportunities.

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“…Especially, the bright fluorescence in CH3 suggests a highly efficient reaction of probe with both H 2 S and phosphatase. To confirm the origin of the fluorescence signals, negative control experiments were performed by employing a phosphatase inhibitor (Na 3 VO 4 ) and an H 2 S‐generated enzyme inhibitor (aminooxyacetic acid, AOAA) . After the cells were sequentially treated with Na 3 VO 4 and the probe, the fluorescences in CH2 and CH3 from phosphatase obviously reduced, but the fluorescent signal in CH1 from H 2 S became slightly stronger due to the interruption of FRET when phosphatase activity had been inhibited greatly (middle column of Figure a).…”

Section: Figurementioning

confidence: 99%

Gasotransmitter Regulation of Phosphatase Activity in Live Cells Studied by Three‐Channel Imaging Correlation

Zhang

Liu

et al. 2019

Angewandte Chemie

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Enzyme activity in live cells is dynamically regulated by small‐molecule transmitters for maintaining normal physiological functions. A few probes have been devised to measure intracellular enzyme activities by fluorescent imaging, but the study of the regulation of enzyme activity via gasotransmitters in situ remains a long‐standing challenge. Herein, we report a three‐channel imaging correlation by a single dual‐reactive fluorescent probe to measure the dependence of phosphatase activity on the H2S level in cells. The two sites of the probe reactive to H2S and phosphatase individually produce blue and green fluorescent responses, respectively, and resonance energy transfer can be triggered by their coexistence. Fluorescent analysis based on the three‐channel imaging correlation shows that cells have an ideal level of H2S to promote phosphatase activity up to its maximum. Significantly, a slight deviation from this H2S level leads to a sharp decrease of phosphatase activity. The discovery further strengthens our understanding of the importance of H2S in cellular signaling and in various human diseases.

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In Situ Localization of Enzyme Activity in Live Cells by a Molecular Probe Releasing a Precipitating Fluorochrome

Abstract: Supportinginformation and the ORCID identification number(s) for the author(s) of this article can be found under: https://doi.

Cited by 181 publications

References 43 publications

Excited-state intramolecular proton-transfer (ESIPT) based fluorescence sensors and imaging agents

Excited-state intramolecular proton-transfer (ESIPT) based fluorescence sensors and imaging agents

Enzyme-Activated Fluorogenic Probes for Live-Cell and in Vivo Imaging

Gasotransmitter Regulation of Phosphatase Activity in Live Cells Studied by Three‐Channel Imaging Correlation

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