Self‐Immolative Bioluminogenic Quinone Luciferins for NAD(P)H Assays and Reducing Capacity‐Based Cell Viability Assays

Zhou, Wenhui; Leippe, Donna; Duellman, Sarah; Sobol, Mary; Vidugirienė, Jolanta; O’Brien, Martha A.; Shultz, John; Kimball, Joshua J.; DiBernardo, Céline; Moothart, Leonard; Bernad, Laurent; Cali, James J.; Klaubert, Dieter H.; Meisenheimer, Poncho

doi:10.1002/cbic.201300744

Cited by 27 publications

(27 citation statements)

References 56 publications

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“…This wasi na ccordance with the previous studies, indicative of the rapid reduction of the quinone in the presence of NQO1 enzyme;t he peak at 8.3 min corresponded to the formationo f lactone (Figure 2b). [15] The rate of decomposition of the compound was calculated to be 0.063 min À1 which correlatedw ell with the rate of formation of lactone that is, 0.066 min À1 (See SI, Figure S5 Having established the in vitro release of H 2 Sf rom the donor motifs, we next investigatedt he cellular aspects of controlled generation of this reactive gaseous species. Protein persulfidation is ap rotective post-translational modification associated with H 2 Sw herein, thiol of ac ysteine (Cys-SH) reacts with H 2 Si nt he presence of oxidants to form more reactive persulfide (Cys-SSH).…”

Section: Resultsmentioning

confidence: 97%

Carbonyl Sulfide (COS) Donor Induced Protein Persulfidation Protects against Oxidative Stress

Chauhan

Gupta

Ravikumar

et al. 2019

Chemistry An Asian Journal

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The emergence of hydrogen sulfide (H 2 S) as an important signallingm olecule in redox biology with therapeutic potentialh as triggered interesti ng enerating this molecule within cells. One strategy that has been proposed is to use carbonyl sulfide (COS) as as urrogate for hydrogen sulfide. Small molecules that generate COS have been shown to produce hydrogen sulfide in the presenceo fc arbonic anhydrase, aw idely prevalent enzyme. However,o ther studies have indicated that COS may have biological effects which are distinct from H 2 S. Thus, it would be useful to develop tools to compare (and contrast) effects of COS and H 2 S. Here we report enzyme-activated COS donors that are capable of inducing protein persulfidation, which is symptomatic of generation of hydrogen sulfide. The COS donors are also capable of mitigating stress induced by elevated reactive oxygen species. To gether,o ur data suggests that the effects of COS parallel that of hydrogen sulfide,l aying the foundation for further development of these donors as possible therapeutic agents.

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Section: Resultsmentioning

confidence: 97%

Carbonyl Sulfide (COS) Donor Induced Protein Persulfidation Protects against Oxidative Stress

Chauhan

Gupta

Ravikumar

et al. 2019

Chemistry An Asian Journal

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“…[161] Pagel and co-workers [162] have recently developed new NQO1-responsive CEST-MRI agent 124 ( Figure 3) based on quinone-type TML moietiesf or the imaging of enzymatic activity in tissue. [161] Pagel and co-workers [162] have recently developed new NQO1-responsive CEST-MRI agent 124 ( Figure 3) based on quinone-type TML moietiesf or the imaging of enzymatic activity in tissue.…”

Section: Quinone-oxidoreductase-sensitive Tml Systemsmentioning

confidence: 99%

“…Additionally,i n2 014, Zhou and co-workers reported the synthesis ande valuationo fN QO1-sensitive quinone-TML-based pro-biolumigenic luciferin derivatives 119-123 (Scheme 26); these derivatives have successfully been applied for assays detecting NADP(H) or the reducingc apacity of viable cells with very high sensitivity,f ast response, and low background noise. [161] Pagel and co-workers [162] have recently developed new NQO1-responsive CEST-MRI agent 124 (Figure 3) based on quinone-type TML moietiesf or the imaging of enzymatic activity in tissue. In magnetic resonance imaging (MRI), the resonance excitation of protons in tissue is used to generate pictures of the tissue with high resolution.…”

Section: Quinone-oxidoreductase-sensitive Tml Systemsmentioning

confidence: 99%

Trimethyl Lock: A Multifunctional Molecular Tool for Drug Delivery, Cellular Imaging, and Stimuli‐Responsive Materials

Okoh

Klahn

2018

ChemBioChem

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Trimethyl lock (TML) systems are based on ortho-hydroxydihydrocinnamic acid derivatives displaying increased lactonization reactivity owing to unfavorable steric interactions of three pendant methyl groups, and this leads to the formation of hydrocoumarins. Protection of the phenolic hydroxy function or masking of the reactivity as benzoquinone derivatives prevents lactonization and provides a trigger for controlled release of molecules attached to the carboxylic acid function through amides, esters, or thioesters. Their easy synthesis and possible chemical adaption to several different triggers make TML a highly versatile module for the development of drug-delivery systems, prodrug approaches, cell-imaging tools, molecular tools for supramolecular chemistry, as well as smart stimuliresponsive materials.

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“…32 In the presence of NADH or NADPH, the proluciferin is converted to luciferin by a reductase enzyme and the luciferin is used by luciferase to produce light in proportion to the amount of NADH or NADPH. To measure an oxidized dinucleotide, or total dinucleotide in a mixed pool of reduced and oxidized forms, the oxidized form is first reduced by a dehydrogenase enzyme/substrate pair with selectivity for either NAD or NADP (Fig.…”

Section: Assay Principle and Formatsmentioning

confidence: 99%

Bioluminescent Cell-Based NAD(P)/NAD(P)H Assays for Rapid Dinucleotide Measurement and Inhibitor Screening

Vidugirienė

Leippe

Sobol

et al. 2014

ASSAY and Drug Development Technologies

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The central role of nicotinamide adenine dinucleotides in cellular energy metabolism and signaling makes them important nodes that link the metabolic state of cells with energy homeostasis and gene regulation. In this study, we describe the implementation of cellbased bioluminescence assays for rapid and sensitive measurement of those important redox cofactors. We show that the sensitivity of the assays (limit of detection *0.5 nM) enables the selective detection of total amounts of nonphosphorylated or phosphorylated dinucleotides directly in cell lysates. The total amount of NAD+NADH or NADP+NADPH levels can be detected in as low as 300 or 600 cells/ well, respectively. The signal remains linear up to 5,000 cells/well with the maximum signal-to-background ratios ranging from 100 to 200 for NAD+NADH and from 50 to 100 for NADP+NADPH detection. The assays are robust (Z 0 value > 0.7) and the inhibitor response curves generated using a known NAD biosynthetic pathway inhibitor FK866 correlate well with the reported data. More importantly, by multiplexing the dinucleotide detection assays with a fluorescent nonmetabolic cell viability assay, we show that dinucleotide levels can be decreased dramatically (>80%) by FK866 treatment before changes in cell viability are detected. The utility of the assays to identify modulators of intracellular nicotinamide adenine dinucleotide levels was further confirmed using an oncology active compound library, where novel dinucleotide regulating compounds were identified. For example, the histone deacetylase inhibitor entinostat was a potent inhibitor of cellular nicotinamide adenine dinucleotides, whereas the selective estrogen receptor modulator raloxifene unexpectedly caused a twofold increase in cellular nicotinamide adenine dinucleotide levels.

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Self‐Immolative Bioluminogenic Quinone Luciferins for NAD(P)H Assays and Reducing Capacity‐Based Cell Viability Assays

Abstract: Highly sensitive self-cleavable trimethyl lock quinone-luciferin substrates for diaphorase were designed and synthesized to measure NAD(P)H in biological samples and monitor viable cells via NAD(P)H-dependent cellular oxidoreductase enzymes and their NAD(P)H cofactors.

Cited by 27 publications

References 56 publications

Carbonyl Sulfide (COS) Donor Induced Protein Persulfidation Protects against Oxidative Stress

Carbonyl Sulfide (COS) Donor Induced Protein Persulfidation Protects against Oxidative Stress

Trimethyl Lock: A Multifunctional Molecular Tool for Drug Delivery, Cellular Imaging, and Stimuli‐Responsive Materials

Bioluminescent Cell-Based NAD(P)/NAD(P)H Assays for Rapid Dinucleotide Measurement and Inhibitor Screening

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