Unprecedented Sensitivity in a Probe for Monitoring Cathepsin B: Chemiluminescence Microscopy Cell‐Imaging of a Natively Expressed Enzyme

Roth-Konforti, Michal E.; Bauer, Christoph; Shabat, Doron

doi:10.1002/anie.201709347

Cited by 145 publications

(113 citation statements)

References 36 publications

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“…In aqueous media, these next‐generation phenoxy‐dioxetane chemiluminescent luminophores exhibit up to 3000‐fold greater light emission than the original Schaap's dioxetanes . This discovery has already been exploited in a number of applications that require rapid and sensitive detection of various enzymes and analytes . In addition, the intense light emitted from the probe can be used to effectively image transfected and native enzymatic activity in live cells .…”

Section: Introductionmentioning

confidence: 99%

Chemiluminescent Carbapenem‐Based Molecular Probe for Detection of Carbapenemase Activity in Live Bacteria

Das

Ihssen²,

Wick³

et al. 2020

Chemistry A European J

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Carbapenemase‐producing organisms (CPOs) pose a severe threat to antibacterial treatment due to the acquisition of antibiotic resistance. This resistance can be largely attributed to the antibiotic‐hydrolyzing enzymes that the bacteria produce. Current carbapenem “wonder drugs”, such as doripenem, ertapenem, meropenem, imipenem, and so on, are resistant to regular β‐lactamases, but susceptible to carbapenemases. Even worse, extended exposure of bacteria to these drugs accelerates the spread of resistance genes. In order to preserve the clinical efficacy of antibacterial treatment, carbapenem drugs should be carefully regulated and deployed only in cases of a CPO infection. Early diagnosis is therefore of paramount importance. Herein, we report the design, synthesis, and activity of the first carbapenemase‐sensitive chemiluminescent probe, CPCL, which may be used to monitor CPO activity. The design of our probe enables enzymatic cleavage of the carbapenem core, which is followed by a facile 1,8‐elimination process and the emission of green light through rapid chemical excitation. We have demonstrated the ability of the probe to detect a number of clinically relevant carbapenemases and the successful identification of CPO present in bacterial cultures, such as those used for clinical diagnosis. We believe that our use of “turn‐on” chemiluminescence activation will find significant application in future diagnostic assays and improve antibacterial treatment.

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Section: Introductionmentioning

confidence: 99%

Chemiluminescent Carbapenem‐Based Molecular Probe for Detection of Carbapenemase Activity in Live Bacteria

Das

Ihssen²,

Wick³

et al. 2020

Chemistry A European J

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“…Presumptive colonies have to be confirmed after the initial isolation. [25][26][27][28][29][30][31][32][33][34] Ar emarkable enhancement of light emission was obtained by simply improving the emissive nature of the excited species,f ormed during the chemiexcitation of Schaapsdioxetanes. Positive results are available after 96-144 h(4-6 days), depending on the growth of the bacteria.…”

mentioning

confidence: 99%

Ultrasensitive Detection of Salmonella and Listeria monocytogenes by Small‐Molecule Chemiluminescence Probes

et al. 2019

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Detection of Salmonella and L. monocytogenes in food samples by current diagnostic methods requires relatively long time to results (2-6 days). Furthermore,t he ability to perform environmental monitoring at the factory site for these pathogens is limited due to the need for laboratory facilities. Herein, we report new chemiluminescence probes for the ultrasensitive direct detection of viable pathogenic bacteria. The probes are composed of ab right phenoxy-dioxetane luminophore masked by triggering group,whichisactivated by aspecific bacterial enzyme,and could detect their corresponding bacteria with an LOD value of about 600-fold lower than that of fluorescent probes.M oreover,w ew ere able to detect am inimum of 10 Salmonella cells within 6hincubation. The assayallows for bacterial enrichment and detection in one test tube without further sample preparation. We anticipate that this design strategy will be used to prepare analogous chemiluminescence probes for other enzymes relevant to specific bacteria detection and point-of-care diagnostics.

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“…For the design of protein‐based Zn 2+ sensors, the CFP donor and YFP acceptor were linked by a domain that selectively recognized and bound Zn 2+ ions, leading to changes of both conformation and FRET efficiency (Figure c) . In addition, a dioxetane‐based chemiluminescence probe was also designed for the detection of cathepsin B enzyme; a lysosomal cysteine protease overexpressed in specific types of cancers (Figure d) ,…”

Section: Fluorogenic Bifunctional Linkersmentioning

confidence: 99%

Emerging Applications of Fluorogenic and Non‐fluorogenic Bifunctional Linkers

Liu

2018

Chemistry A European J

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Homo- and hetero-bifunctional linkers play vital roles in constructing a variety of functional systems, ranging from protein bioconjugates with drugs and functional agents, to surface modification of nanoparticles and living cells, and to the cyclization/dimerization of synthetic polymers and biomolecules. Conventional approaches for assaying conjugation extents typically rely on ex situ techniques, such as mass spectrometry, gel electrophoresis, and size-exclusion chromatography. If the conjugation process involving bifunctional linkers was rendered fluorogenic, then in situ monitoring, quantification, and optical tracking/visualization of relevant processes would be achieved. In this review, conventional non-fluorogenic linkers are first discussed. Then the focus is on the evolution and emerging applications of fluorogenic bifunctional linkers, which are categorized into hetero-bifunctional single-caging fluorogenic linkers, homo-bifunctional double-caging fluorogenic linkers, and hetero-bifunctional double-caging fluorogenic linkers. In addition, stimuli-cleavable bifunctional linkers designed for both conjugation and subsequent site-specific triggered release are also summarized.

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Unprecedented Sensitivity in a Probe for Monitoring Cathepsin B: Chemiluminescence Microscopy Cell‐Imaging of a Natively Expressed Enzyme

Cited by 145 publications

References 36 publications

Chemiluminescent Carbapenem‐Based Molecular Probe for Detection of Carbapenemase Activity in Live Bacteria

Chemiluminescent Carbapenem‐Based Molecular Probe for Detection of Carbapenemase Activity in Live Bacteria

Ultrasensitive Detection of Salmonella and Listeria monocytogenes by Small‐Molecule Chemiluminescence Probes

Emerging Applications of Fluorogenic and Non‐fluorogenic Bifunctional Linkers

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