Inhibiting Firefly Bioluminescence by Chalcones

Zhang, Huateng; Su, Jing; Lin, Yuxin; Bai, Haixiu; Liu, Jiaxiang; Chen, Hui; Du, Lüpei; Gu, Lichuan; Li, Minyong

doi:10.1021/acs.analchem.7b00813

Cited by 16 publications

(18 citation statements)

References 39 publications

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“…As shown in Figure 6A , in 50–200 μM range a complete inhibition on hSIRT1 by both resveratrol and quercetin was observed when the bioluminescent assay was applied. A complete inhibition was observed also by using the non-acetylated control substrate (data not shown), thus allowing to exclude a direct inhibition of resveratrol and quercetin on hSIRT1; conversely, the observed effect can depend on the capability of resveratrol and flavonoids to inhibit the firefly luciferase activity ( Bakhtiarova et al, 2006 ; Auld et al, 2008 ; Zhang et al, 2017 ). A similar behavior was observed also for WM-SRT ( Figure 6B ).…”

Section: Resultsmentioning

confidence: 86%

“…These compounds have been reported as potent hSIRT1 activators ( Howitz et al, 2003 ), but with respect to their activation mechanism inconsistent and conflicting findings have been reported ( Hubbard and Sinclair, 2014 ). The combinatory use of bioluminescence/HTRF ® sirtuin assays proved to be crucial in identifying the false inhibition effect of both tested compounds shown by SIRT-Glo TM assay, which actually may be attributed to the capability of resveratrol and flavonoids to inhibit the firefly luciferase activity ( Bakhtiarova et al, 2006 ; Auld et al, 2008 ; Zhang et al, 2017 ). Interestingly, by using HTRF ® assay no effect of resveratrol was observed on both WM-SRT and hSIRT1 activities, while only a slight increase, up to about 20%, of hSIRT1 activity by quercetin was observed, much lower than that reported in previous studies using Fluor de Lys assay ( Howitz et al, 2003 ; de Boer et al, 2006 ).…”

Section: Discussionmentioning

confidence: 99%

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Measuring Activity of Native Plant Sirtuins - The Wheat Mitochondrial Model

et al. 2018

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Sirtuins are NAD+-dependent deacetylase enzymes that have gained considerable interest in mammals for their recognized importance in gene silencing and expression and in cell metabolism. Conversely, knowledge about plant sirtuins remains limited, although a sirtuin-mediated regulation of mitochondrial energy metabolism has been recently reported in Arabidopsis. However, so far, no information is available about direct measurement of intracellular plant sirtuin activity, i.e., in cell extracts and/or subcellular organelles. In this study, a novel approach was proposed for reliable evaluation of native sirtuin activity in plant samples, based on (i) an adequate combinatory application of enzymatic assays very different for chemical basis and rationale and (ii) a comparative measurement of activity of a recombinant sirtuin isoform. In particular, two sirtuin assays were applied, based on bioluminescence emission and Homogeneous Time-Resolved Fluorescence (HTRF®) technology, and the human SIRT1 isoform (hSIRT1) was used for comparison. For the first time in plants, this new approach allowed measuring directly a high and nicotinamide-sensitive sirtuin activity in highly purified mitochondrial fraction obtained from durum wheat (WM). WM-sirtuin activity was 268 ± 10 mU⋅mg-1 protein, as measured by HTRF® assay, and 166 ± 12 ng hSIRT1 eq.⋅mg-1 protein, as evaluated by the bioluminescent assay and calculated on the basis of the hSIRT1 calibration curve. Moreover, effects of resveratrol and quercetin, reported as potent hSIRT1 activators, but whose activation mechanism is still debated, were also studied. No effect of resveratrol was found on both WM-sirtuin and hSIRT1 activities, while only a slight increase, up to about 20%, of hSIRT1 activity by quercetin was observed. In the whole, results of this study indicate that WM may represent a good system for studying native plant sirtuins. In fact, the high yield of purified WM and their high sirtuin activity, together with use of microplate readers, allow performing a large number of measurements from the same preparation, so qualifying the approach for application to large-scale high-throughput screening. Moreover, WM may also represent an excellent tool to investigate physiological role and modulation of plant sirtuins under experimental conditions more physiologically relevant with respect to recombinant purified enzymes.

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Section: Resultsmentioning

confidence: 86%

Section: Discussionmentioning

confidence: 99%

Measuring Activity of Native Plant Sirtuins - The Wheat Mitochondrial Model

et al. 2018

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“…In 2017, Zhang et al found that a series of chalcone compounds were identified as Fluc inhibitors with favorable potency both in vitro and in vivo. 11 Resveratrol, which shows therapeutic potential in patients with cancer, diabetes, and Alzheimer's disease, has been identified as a potent inhibitor of FLuc (K i = 2 μM). 12 Resveratrol's activity reported by most studies is basically at the level of micromolar, but many of these studies utilized the FLuc assay.…”

Section: ■ Introductionmentioning

confidence: 99%

“…For example, chalcone refers to an aromatic ketone and an enone which has exhibited a broad range of therapeutic activity, such as anticancer, antiparasitic, anti-HIV, and so on. In 2017, Zhang et al found that a series of chalcone compounds were identified as Fluc inhibitors with favorable potency both in vitro and in vivo . Resveratrol, which shows therapeutic potential in patients with cancer, diabetes, and Alzheimer’s disease, has been identified as a potent inhibitor of FLuc ( K i = 2 μM) .…”

Section: Introductionmentioning

confidence: 99%

Structural Analysis and Identification of False Positive Hits in Luciferase-Based Assays

Yang

Dong

Yang

et al. 2020

J. Chem. Inf. Model.

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Luciferase-based bioluminescence detection techniques are highly favored in high-throughput screening (HTS), in which the firefly luciferase (FLuc) is the most commonly used variant. However, FLuc inhibitors can interfere with the activity of luciferase, which may result in false positive signals in HTS assays. In order to reduce the unnecessary cost of time and money, an in silico prediction model for FLuc inhibitors is highly desirable. In this study, we built an extensive data set consisting of 20 888 FLuc inhibitors and 198 608 noninhibitors, and then developed a group of classification models based on the combination of three machine learning (ML) algorithms and four types of molecular representations. The best prediction model based on XGBoost and ECFP4 and MOE2d descriptors yielded a balanced accuracy (BA) of 0.878 and an area under the receiver operating characteristic curve (AUC) value of 0.958 for the validation set, and a BA of 0.886 and an AUC of 0.947 for the test set. Three external validation sets, including set 1 (3231 FLuc inhibitors and 69 783 noninhibitors), set 2 (695 FLuc inhibitors and 75 913 noninhibitors), and set 3 (1138 FLuc inhibitors and 8155 noninhibitors), were used to verify the predictive ability of our models. The BA values for the three external validation sets given by the best model are 0.864, 0.845, and 0.791, respectively. In addition, the important features or structural fragments related to FLuc inhibitors were recognized by the Shapley additive explanations (SHAP) method along with their influences on predictions, which may provide valuable clues to detecting undesirable luciferase inhibitors. Based on the important and explanatory features, 16 rules were proposed for detecting FLuc inhibitors, which can achieve a correction rate of 70% for FLuc inhibitors. Furthermore, a comparison with existing prediction rules and models for FLuc inhibitors used in virtual screening verified the high reliability of the models and rules proposed in this study. We also used the model to screen three curated chemical databases, and almost 10% of the molecules in the evaluated databases were predicted as inhibitors, highlighting the potential risk of false positives in luciferase-based assays. Finally, a public web server called ChemFLuc was developed (), and it offers a free available service to predict potential FLuc inhibitors.

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“…The naturally available chalcones play a beneficial role in anti‐cancer activity, bioluminescence and active plant resistance of biological applications. However, the synthetic chalcone monomers have attractive features in host–guest chemistry and optical applications.…”

Section: Introductionmentioning

confidence: 99%

Acetophenone and thiophene side‐arm polyaryleneethynylene conjugated polymers for enrichment of electronic applications

Kumar

Chandramohan²,

Priya

et al. 2020

Polymer International

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In the present work, a new type of semiconducting acetophenone and thiophene side‐arm based polyaryleneethynylene (PAE) conjugated polymers (CPs) were synthesized using the Heck–Sonogashira cross‐coupling reaction. The new type of monomers and APAEC8, APAEC12, APAEF8, TPAEC8, TPAEC12, and TPAEF8 (PAE CPs) were synthesized and characterized using Fourier transform IR, NMR and high resolution mass spectrometry studies. The average molecular weight of the CPs was determined using gel permeation chromatography. The data obtained from optical properties in the UV–visible and photoluminescence spectral studies were used to calculate the values of the Stokes shift. Among the six CPs studied, TPAEC8 was found to possess a lower optical band gap (Eg = 2.112 eV) than those of the other CPs. The data obtained from different studies, it was found that TPAEC8 exhibits better properties than the other CPs and can be used in the field of organic based photonic, electronic and other sensor applications. © 2020 Society of Chemical Industry

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Inhibiting Firefly Bioluminescence by Chalcones

Cited by 16 publications

References 39 publications

Measuring Activity of Native Plant Sirtuins - The Wheat Mitochondrial Model

Measuring Activity of Native Plant Sirtuins - The Wheat Mitochondrial Model

Structural Analysis and Identification of False Positive Hits in Luciferase-Based Assays

Acetophenone and thiophene side‐arm polyaryleneethynylene conjugated polymers for enrichment of electronic applications

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