Imaging of Enzyme Activity by Electron Paramagnetic Resonance: Concept and Experiment Using a Paramagnetic Substrate of Alkaline Phosphatase

Sanzhaeva, Urikhan; Xu, Xuan; Guggilapu, PriyaankaDevi; Tseytlin, Mark; Khramtsov, Valery V.; Driesschaert, Benoît

doi:10.1002/ange.201806851

Cited by 2 publications

(1 citation statement)

References 27 publications

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“…Of course, substrates for use with other approaches are available too. These include bioluminescence, [32] electrochemistry, [33] UV‐Vis spectroscopy, [34] Raman spectroscopy and surface‐enhanced (resonance) Raman spectroscopy (SERS and SERRS), [35] chemical exchange saturation transfer magnetic resonance imaging (CEST MRI), [36] electron paramagnetic resonance (EPR) spectroscopy, [37] and 19 F nuclear magnetic resonance (NMR) spectroscopy [38] …”

Section: Detection Methodsmentioning

confidence: 99%

Methods to Characterise Enzyme Kinetics with Biological and Medicinal Substrates: The Case of Alkaline Phosphatase

Harroun

Vallée‐Bélisle

2023

Chemistry Methods

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Alkaline phosphatase (AP) enzymes are of broad interest in fields ranging from biochemistry and medicine to biotechnology and nanotechnology. Characterising the catalytic activity of AP is typically realised by either employing non‐natural signal‐generating substrates that are detectable by absorbance and fluorescence spectroscopy or by quantifying the release of inorganic phosphate by the classic malachite green assay. The latter method is often required for studying “spectroscopically silent” biomolecular substrates, but it does not enable continuous monitoring of kinetics in real‐time. In recent years, newer techniques for studying AP function have been developed to circumvent this limitation, including fluorescent and colourimetric substrate‐specific assays based on supramolecular chemistry, organic probes and nanomaterials, as well as other assays based on isothermal titration calorimetry, direct detection with infrared spectroscopy and mass spectrometry, and monitoring conformational change by fluorescent nanoantennas. Here, we review these strategies and comment on their strengths and weaknesses in the context of AP.

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Section: Detection Methodsmentioning

confidence: 99%

Methods to Characterise Enzyme Kinetics with Biological and Medicinal Substrates: The Case of Alkaline Phosphatase

Harroun

Vallée‐Bélisle

2023

Chemistry Methods

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Highly Sensitive “Off/On” EPR Probes to Monitor Enzymatic Activity

Elkhanoufi

Stefanìa

Alberti

et al. 2022

Chemistry A European J

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The assessment of unregulated level of enzyme activity is a crucial parameter for early diagnoses in a wide range of pathologies. In this study, we propose the use of electron paramagnetic resonance (EPR) as an easy method to probe carboxylesterase (CE) enzymatic activity in vitro. For this application, were synthesized two amphiphilic, nitroxide containing esters, namely Tempo‐C12 (T‐C12) and Tempo‐2‐C12 (T‐2‐C12). They exhibit low solubility in water and form stable micelles in which the radicals are EPR almost silent, but the hydrolysis of the ester bond yields narrows and intense EPR signals. The intensity of the EPR signals is proportional to the enzymatic activity. CEs1, CEs2 and esterase from porcine liver (PLE) were investigated. The obtained results show that T‐C12 and T‐2‐C12‐containing systems display a much higher selectivity toward the CEs2, with a Limit of Detection of the same order of those ones obtained with optical methods.

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Imaging of Enzyme Activity by Electron Paramagnetic Resonance: Concept and Experiment Using a Paramagnetic Substrate of Alkaline Phosphatase

Cited by 2 publications

References 27 publications

Methods to Characterise Enzyme Kinetics with Biological and Medicinal Substrates: The Case of Alkaline Phosphatase

Methods to Characterise Enzyme Kinetics with Biological and Medicinal Substrates: The Case of Alkaline Phosphatase

Highly Sensitive “Off/On” EPR Probes to Monitor Enzymatic Activity

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