Protein sensing in living cells by molecular rotor-based fluorescence-switchable chemical probes

Yu, Wan-Ting; Wu, Ting-Wei; Huang, Chi-Ling; Chen, I-Chia; Tan, Kui‐Thong

doi:10.1039/c5sc02808f

Cited by 83 publications

(54 citation statements)

References 43 publications

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“…During the completion of this manuscript, Yu et al . [ 38 ] reported BG modified with a fluorescent molecular rotor that increased in fluorescence upon labeling MGMT. However, fluorophore-labeled BG (or SNAP-tag probes) are poor substrates for wild-type MGMT [ 19 , 39 ] and as a consequence the fluorescence response with enzyme was slow, requiring high concentrations of probe (5 μM) to produce signals on a timescale of hours.…”

Section: Discussionmentioning

confidence: 99%

Fluorogenic Real-Time Reporters of DNA Repair by MGMT, a Clinical Predictor of Antitumor Drug Response

et al. 2016

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Common alkylating antitumor drugs, such as temozolomide, trigger their cytotoxicity by methylating the O6-position of guanosine in DNA. However, the therapeutic effect of these drugs is dampened by elevated levels of the DNA repair enzyme, O6-methylguanine DNA methyltransferase (MGMT), which directly reverses this alkylation. As a result, assessing MGMT levels in patient samples provides an important predictor of therapeutic response; however, current methods available to measure this protein are indirect, complex and slow. Here we describe the design and synthesis of fluorescent chemosensors that report directly on MGMT activity in a single step within minutes. The chemosensors incorporate a fluorophore and quencher pair, which become separated by the MGMT dealkylation reaction, yielding light-up responses of up to 55-fold, directly reflecting repair activity. Experiments show that the best-performing probe retains near-native activity at mid-nanomolar concentrations. A nuclease-protected probe, NR-1, was prepared and tested in tumor cell lysates, demonstrating an ability to evaluate relative levels of MGMT repair activity in twenty minutes. In addition, a probe was employed to evaluate inhibitors of MGMT, suggesting utility for discovering new inhibitors in a high-throughput manner. Probe designs such as that of NR-1 may prove valuable to clinicians in selection of patients for alkylating drug therapies and in assessing resistance that arises during treatment.

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Section: Discussionmentioning

confidence: 99%

Fluorogenic Real-Time Reporters of DNA Repair by MGMT, a Clinical Predictor of Antitumor Drug Response

et al. 2016

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“…The development of novel organic fluorophores is of great interest in the areas of biological labels and probes, 1 chemosensors, 2 medical diagnostics 3 and photoactive materials. 4 Indeed, the rational design of organic fluorophores has provided a straightforward method for generating various fluorescent functional materials.…”

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confidence: 99%

Rational design of substituted maleimide dyes with tunable fluorescence and solvafluorochromism

et al. 2018

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“…The quantum yield of CQ in PBS buffer (10 mM, pH = 7.4) was 0.08, which increased to 0.36 upon binding to Aβ42 fibrillar aggregates. The observed fluorescence enhancement of CQ can be attributed to the restricted torsional motion of the probe upon binding to Aβ fibrillar aggregates that restricts molecular relaxation, enhances molecular planarity and decreases non-radiative decay rate, resulting in CQ fluorescence enhancement (Yu et al, 2016). To validate the role of restricted molecular motion on the fluorescence enhancement of probe CQ , we recorded the fluorescence spectra of CQ (5 µM, 10 mM PBS) as a function of increased glycerol percentage in the sample solution (Fig.…”

Section: Resultsmentioning

confidence: 99%

Aβ plaque-selective NIR fluorescence probe to differentiate Alzheimer's disease from tauopathies

Rajasekhar

Narayanaswamy

Murugan

et al. 2017

Biosensors and Bioelectronics

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Selective detection and staining of toxic amyloid plaques, a potential biomarker present in the Alzheimer's disease (AD) brain is crucial for both clinical diagnosis and monitoring AD disease progression. Herein, we report a coumarin-quinoline (CQ) conjugate-based turn-on near-infrared (NIR) fluorescence probe for specific detection of β-amyloid (Aβ) aggregates. CQ probe is highly sensitive and exhibits ~100-fold fluorescence enhancement in vitro upon binding Aβ aggregates with enhanced quantum yield. Furthermore, the probe has ~10-fold higher binding affinity towards Aβ aggregates (86nM) compared to commonly used Thioflavin T. Most importantly, CQ probe displays unambiguous selectivity towards Aβ aggregates compared to other toxic protein aggregates such as tau, α-synuclein (α-Syn) and islet amyloid polypeptide (IAPP). In addition, CQ is nontoxic to neuronal cells and shows significant blood brain barrier permeability. Remarkably, CQ stains Aβ plaques in human brain tissue over co-existing tau aggregates and neurofibrillary tangles (NFTs), which are associated in AD and tauopathies. This is a highly desirable attribute to distinguish AD from tau pathology and mixed dementia.

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Protein sensing in living cells by molecular rotor-based fluorescence-switchable chemical probes

Cited by 83 publications

References 43 publications

Fluorogenic Real-Time Reporters of DNA Repair by MGMT, a Clinical Predictor of Antitumor Drug Response

Fluorogenic Real-Time Reporters of DNA Repair by MGMT, a Clinical Predictor of Antitumor Drug Response

Rational design of substituted maleimide dyes with tunable fluorescence and solvafluorochromism

Aβ plaque-selective NIR fluorescence probe to differentiate Alzheimer's disease from tauopathies

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