- A simple bioluminescent method for measuring d-amino acid oxidase activity
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d-Amino acid oxidase (DAO) plays important roles in regulating d-amino acid neurotransmitters and was recently identified as a key enzyme integral to hydrogen sulfide production from d-Cys. We report here the development of a simple biocompatible, bioluminescent method for measuring DAO activity based on the highly selective condensation of d-Cys with 6-hydroxy-2-cyanobenzothiazole (CBT-OH) to form d-luciferin. This journal is
- Bailey, T. Spencer,Donor, Micah T.,Naughton, Sean P.,Pluth, Michael D.
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- A bioluminescent probe for longitudinal monitoring of mitochondrial membrane potential
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Mitochondrial membrane potential (ΔΨm) is a universal selective indicator of mitochondrial function and is known to play a central role in many human pathologies, such as diabetes mellitus, cancer and Alzheimer’s and Parkinson’s diseases. Here, we report the design, synthesis and several applications of mitochondria-activatable luciferin (MAL), a bioluminescent probe sensitive to ΔΨm, and partially to plasma membrane potential (ΔΨp), for non-invasive, longitudinal monitoring of ΔΨm in vitro and in vivo. We applied this new technology to evaluate the aging-related change of ΔΨm in mice and showed that nicotinamide riboside (NR) reverts aging-related mitochondrial depolarization, revealing another important aspect of the mechanism of action of this potent biomolecule. In addition, we demonstrated application of the MAL probe for studies of brown adipose tissue (BAT) activation and non-invasive in vivo assessment of ΔΨm in animal cancer models, opening exciting opportunities for understanding the underlying mechanisms and for discovery of effective treatments for many human pathologies. [Figure not available: see fulltext.]
- Bazhin, Arkadiy A.,Sinisi, Riccardo,De Marchi, Umberto,Hermant, Aurélie,Sambiagio, Nicolas,Maric, Tamara,Budin, Ghyslain,Goun, Elena A.
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- A Convenient Synthetic Method of 2-Cyano-6-methoxybenzothiazole, - A Key Intermediate for the Synthesis of Firefly Luciferin
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The title compound was prepared in one step from commercially available 2-amino-6-methoxybenzothiazole by using the Sandmeyer cyanation reaction.The result enabled us to synthesize firefly luciferin effectively from this amino compound through three steps in 36percent overall yield.
- Toya, Yoshiaki,Takagi, Masaharu,Nakata, Hisao,Suzuki, Nobutaka,Isobe, Minoru,Goto, Toshio
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- Highly Sensitive Bioluminescent Probe for Thiol Detection in Living Cells
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The sensitive detection of thiols including glutathione and cysteine is desirable owing to their roles as indispensable biomolecules in maintaining intracellular biological redox homeostasis. Herein, we report the design and synthesis of SEluc-1 (sulfinate ester luciferin), a chemoselective probe exhibiting a ratiometric and turn-on response towards thiols selectively in fluorescence and bioluminescence, respectively. The probe, which was designed based on the “caged” luciferin strategy, displays excellent selectivity, high signal/noise ratio (>240 in the case of bioluminescence), and a biologically relevant limit of detection (LOD, 80 nm for cysteine), which are all desirable traits for a sensitive bioluminescent sensor. SEluc-1 was further applied to fluorescence imaging of thiol activity in living human cervical cancer HeLa cell cultures, and was successfully able to detect fluctuations in thiol concentrations induced by oxidative stress in a bioluminescent assay utilizing African green monkey fibroblast COS-7 cells and human breast adenocarcinoma MCF-7 cells.
- Hemmi, Mayu,Ikeda, Yuma,Shindo, Yutaka,Nakajima, Takahiro,Nishiyama, Shigeru,Oka, Kotaro,Sato, Moritoshi,Hiruta, Yuki,Citterio, Daniel,Suzuki, Koji
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- Specific Imaging of Tyrosinase in Vivo with 3-Hydroxybenzyl Caged D -Luciferins
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Tyrosinase (TYR), a key enzyme in biosynthesis of melanin, usually functions as a biomarker of severe skin diseases such as vitiligo and melanoma cancer. Accurate detection of TYR activity in vivo is urgent but still challenging. Inspired by the advantages of bioluminescence in vivo strategy in imaging and the specific hydroxylation of 3-hydroxybenzyloxy group by TYR, a bioluminogenic probe, TYR-LH2, was designed and synthesized through caging D-luciferin with 3-hydroxybenzyl. The probe exhibits high selectivity and sensitivity toward TYR with a detection limit of 0.11 U/mL in a small detection volume of 100 μL. Bioluminescence imaging results show that TYR-LH2 is fully competent for monitoring the dynamic changes of TYR in living cells and model animals and possesses the capability of discriminating melanocytes from other cell lines, thus offering a promising approach for investigation and diagnosis of melanoma cancer and other TYR-related diseases in vivo.
- Li, Shuang,Hu, Rui,Wang, Shuangqing,Guo, Xudong,Zeng, Yi,Li, Yi,Yang, Guoqiang
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- Real-Time Bioluminescence Imaging of Nitroreductase in Mouse Model
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Nitroreductase (NTR) is an endogenous reductase overexpressed in hypoxic tumors; however, its precise detection in living cells and animals remains a considerable challenge. Herein, we developed three reaction-based probes and a related bioluminescence assay for the real-time NTR detection. The high sensitivity and selectivity of probe 3, combined with its remarkable potential of bioluminescence imaging, affords a valuable approach for in vivo imaging of NTR in a tumor model mouse.
- Feng, Ping,Zhang, Huateng,Deng, Quankun,Liu, Wei,Yang, Linghui,Li, Guobo,Chen, Guo,Du, Lupei,Ke, Bowen,Li, Minyong
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- A bioluminescent probe for imaging endogenous hydrogen polysulfides in live cells and a murine model of bacterial infection
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In this work, we report the first bioluminescent probe BP-PS for detecting H2Sn with high specificity and sensitivity. Owing to the bioluminescence imaging without requiring an excitation light source, tissue autofluorescence is eliminated and BP-PS shows a high signal-to-noise ratio. Moreover, BP-PS was successfully utilized to visualize endogenous H2Sn in live cells and a murine model of bacterial infection.
- Li, Jun-Bin,Wang, Qianqian,Liu, Hong-Wen,Yuan, Lin,Zhang, Xiao-Bing
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- Real-Time Imaging and Quantification of Peptide Uptake in Vitro and in Vivo
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Peptides constitute an important class of drugs for the treatment of multiple metabolic, oncological, and neurodegenerative diseases, and several hundred novel therapeutic peptides are currently in the preclinical and clinical stages of development. However, many leads fail to advance clinically because of poor cellular membrane and tissue permeability. Therefore, assessment of the ability of a peptide to cross cellular membranes is critical when developing novel peptide-based therapeutics. Current methods to assess peptide cellular permeability are limited by multiple factors, such as the need to introduce rather large modifications (e.g., fluorescent dyes) that require complex chemical reactions as well as an inability to provide kinetic information on the internalization of a compound or distinguish between internalized and membrane-bound compounds. In addition, many of these methods are based on end point assays and require multiple sample manipulation steps. Herein, we report a novel Split Luciferin Peptide (SLP) assay that enables the real-time noninvasive imaging and quantification of peptide uptake both in vitro and in vivo using a very sensitive bioluminescence readout. This method is based on a straightforward, stable chemical modification of the peptide of interest with a d-cysteine tag that preserves the overall peptidic character of the original molecule. This method can be easily adapted for screening peptide libraries and can thus become an important tool for preclinical peptide drug development.
- Karatas, Hacer,Maric, Tamara,D'Alessandro, Pier Luca,Yevtodiyenko, Aleksey,Vorherr, Thomas,Hollingworth, Gregory J.,Goun, Elena A.
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- Highly sensitive and selective bioluminescence based ozone probes and their applications to detect ambient ozone
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Highly selective and sensitive bioluminescence based probes, which respond to ozone but not to other ROS, have been developed. These probes were used to determine ozone concentrations in environmental samples.
- Nam, Younseok,Kim, Beom Seok,Shin, Injae
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- Bioluminescent Turn-On Probe for Sensing Hypochlorite in Vitro and in Tumors
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Hypochlorite (ClO-) is one of the most important reactive oxygen species but using a BL probe for its selective detection (or imaging) still remains challenging. Herein, we report a latent BL probe benzoylhydrazine luciferin (1) for highly selective detection of ClO- in vitro and imaging ClO- in living cells and tumors. In vitro tests indicated that 1 could be applied for highly selective detection of ClO- within the range of 0-62.5 μM with a limit of detection of 0.705 μM. Using these unique features of 1, we successfully applied it to image ClO- in living cells and tumors. We envision that probe 1 might be applied to elucidate the biological roles of ClO- in wider physiological and pathological processes in the near future.
- Chen, Peiyao,Zheng, Zhen,Zhu, Yunxia,Dong, Yu,Wang, Fuqiang,Liang, Gaolin
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- Computation-Guided Rational Design of a Peptide Motif That Reacts with Cyanobenzothiazoles via Internal Cysteine-Lysine Relay
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Site-selective protein modification based on covalent reactions of peptide tags and small molecules is a key capability for basic research as well as for the development of new therapeutic bioconjugates. Here, we describe the computation-guided rational design of a cysteine- and lysine-containing 11-residue peptide sequence that reacts with 2-cyanobenzothiazole (CBT) derivatives. Our data show that the cysteine residue reversibly reacts with the nitrile group on the CBT moiety to form an intermediate thioimidate, which undergoes irreversible SN transfer to the lysine residue, yielding an amidine-linked product. The concepts outlined herein lay a foundation for future development of peptide tags in the context of site-selective modification of lysine residues within engineered microenvironments.
- Keyser, Samantha G. L.,Utz, Ashley,Bertozzi, Carolyn R.
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- A dual-color far-red to near-infrared firefly luciferin analogue designed for multiparametric bioluminescence imaging
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Red-shifted bioluminescent emitters allow improved in vivo tissue penetration and signal quantification, and have led to the development of beetle luciferin analogues that elicit red-shifted bioluminescence with firefly luciferase (Fluc). However, unlike natural luciferin, none have been shown to emit different colors with different luciferases. We have synthesized and tested the first dual-color, far-red to nearinfrared (nIR) emitting analogue of beetle luciferin, which, akin to natural luciferin, exhibits pH dependent fluorescence spectra and emits bioluminescence of different colors with different engineered Fluc enzymes. Our analogue produces different far-red to nIR emission maxima up to λmax=706 nm with different Fluc mutants. This emission is the most redshifted bioluminescence reported without using a resonance energy transfer acceptor. This improvement should allow tissues to be more effectively probed using multiparametric deep-tissue bioluminescence imaging.
- Jathoul, Amit P.,Grounds, Helen,Anderson, James C.,Pule, Martin A.
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- Using Bioluminescence Turn-On to Detect Cysteine in Vitro and in Vivo
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Cysteine (Cys) is an essential amino acid and plays important roles in many biological processes. Bioluminescence (BL) is advantageous in sensitivity but BL probes that were intentionally developed for the selective detection of Cys were rarely reported. Herein, employing a fast conjugate addition between Cys and acrylic ester, we synthesized a caged BL probe acrylic ester luciferin (1) and used it to selectively detect Cys in vitro and image Cys in living cells and in tumor sites. We envision that, in the future, probe 1 might be used for evaluating the Cys roles in more biological processes.
- Zhang, Miaomiao,Wang, Lin,Zhao, Yangyang,Wang, Fuqiang,Wu, Jindao,Liang, Gaolin
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- In Vivo Bioluminescence Imaging of Cobalt Accumulation in a Mouse Model
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As a trace element nutrient, cobalt is critical for both prokaryotes and eukaryotes. In the current study, a turn-on Cobalt Bioluminescent Probe 1 (CBP-1) for the detection of cobalt has been successfully developed based on oxidative C-O bond cleavage. This probe exhibited high selectivity and sensitivity toward cobalt over other analytes. By using CBP-1, the successful in vivo imaging of cobalt accumulation was carried out in a mouse model. Such an ability to determine cobalt in living animals provides a powerful technology for studying the system distribution, toxic potency, and biological effect of Co2+.
- Ke, Bowen,Ma, Lin,Kang, Ting,He, Wei,Gou, Xueyan,Gong, Deying,Du, Lupei,Li, Minyong
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- Synthesis and evaluation of D-thioluciferin, a bioluminescent 6'-thio analog of Dluciferin
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All known light-emitting firefly-bioluminescent luciferin analogs are either derived from the 6'-hydroxy- and/or 6'-aminoluciferin. We report the synthesis of D-thioluciferin, a 6'-thio analog or isostere of D-luciferin, starting from p-aminothiophenol, using a unique thioacrylate-S-protecting-group strategy. Upon treatment of Dthioluciferin with purified Photinus pyralis (Ppy) luciferase (Luc), a bioluminescence emission with a red-shift λmax relative to D-luciferin was observed. It was also shown that disulphide and sulphide analogs of Dthioluciferin did not produce similar bioluminescences relative to D-thioluciferin when treated with Ppy Luc under standard conditions, thus, providing a foundation for the development of D-thioluciferin based probes based on disulphide reduction and S-dealkylation.
- Rylands, Marwaan,Jardine, Anwar
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p. 176 - 189
(2021/03/17)
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- Characterization of the reactivity of luciferin boronate - A probe for inflammatory oxidants with improved stability
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Boronate derivatives of luciferin, containing oxidant-activated self-immolative moieties, recently have been developed for bioluminescent detection of hydrogen peroxide in animal models. Here, we report the synthesis and characterization of luciferin boronic acid pinacol ester (LBE) as a probe for detection of hydrogen peroxide, hypochlorous acid, and peroxynitrite, with improved stability and response time. HPLC analyses showed that LBE quickly hydrolyzes in phosphate buffer to luciferin boronic acid (LBA). Hydrogen peroxide oxidizes LBA slowly, with the formation of luciferase substrate, luciferin (Luc-OH), as the only product. Hypochlorite also oxidizes LBA to luciferin, but the subsequent reaction of Luc-OH with hypochlorite gives a chlorinated luciferin Luc–OH–Cl, which has a higher fluorescence quantum yield than luciferin at pH 7.4 and is also a substrate for luciferase (Takakura H, et. all. ChemBioChem 2012; 13:1424). Similar to other boronate probes, LBA is oxidized by peroxynitrite in two pathways. Luc-OH is the product of the major pathway, common for all the oxidants tested, whereas the non-fluorescent nitrated derivative, Luc-NO2, is formed in the minor pathway, specific for peroxynitrite. Formation of luciferin radical intermediate in the minor pathway has been confirmed by EPR spin trapping and mass spectrometric analyses of the spin adducts. We conclude that LBE shows potential as an improved probe for the detection of inflammatory oxidants in biological settings. Complementation of the bioluminescence measurements by HPLC or LC-MS-based identification of chlorinated and nitrated luciferin(s) will help identify the oxidants detected.
- ?wierczyńska, Ma?gorzata,Grzelakowska, Aleksandra,Modrzejewska, Julia,Podsiad?y, Rados?aw,S?owiński, Daniel,Siarkiewicz, Przemys?aw,Szala, Marcin,Zielonka, Jacek
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- DERIVATIVES OF LUCIFERIN AND METHODS FOR THEIR SYNTHESIS
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6-Thio derivatives of D-luciferin, also referred to as D-thioluciferins, having the general structure of Formula (I) are provided. Methods for synthesising D-luciferin, its derivatives, and their related 2-cyanobenzothiazole precursors are also provided. These compounds are commercially valuable due to their application in optical imaging, particularly in bioluminescence imaging.
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- Bioluminescent probe for detection of selenocysteine in organisms, preparation method and application thereof
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Belonging to the field of organic synthesis and detection, the invention relates to a bioluminescent probe for detection of selenocysteine in organisms, a preparation method and application thereof. The bioluminescent probe BF-1 for recognition of Sec provided by the invention has the advantages of good selectivity, high sensitivity, low detection limit (8nM), good biocompatibility, etc. In a Tris-HCl buffer solution, the bioluminescent intensity and selenocysteine concentration show a good linear relationship, thus indicating that the probe is suitable for quantitative detection of selenocysteine. The probe BF-1 also realizes bioluminescence imaging of selenocysteine in MCF-7-luc cells. The probe BF-1 also achieves sensitive detection of endogenous Sec level in vivo (FVB-Luc transgenicmice), also is quick in response, and the bioluminescence signal is stable. The probe BF-1 prepared according to the invention is an effective tool for visual and quantitative detection of the selenocysteine content in cells, living bodies and tumor tissues.
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- A Chemo-Enzymatic Cascade for the Smart Detection of Nitro- and Halogenated Phenols
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The flavin-dependent monooxygenase, HadA, catalyzes the dehalogenation and denitration of the toxicants, nitro- and halogenated phenols, to benzoquinone. The HadA reaction can be applied in one-pot reactions towards the de novo synthesis of d-luciferin by coupling with d-Cys condensation. d-luciferin, a valuable chemical widely used in biomedical applications, can be used as a substrate for the reaction of firefly luciferase to generate bioluminescence. As nitro- and halogenated phenols are key indicators of human overexposure to pesticides and pesticide contamination, the technology provides a sensitive and convenient tool for improved biomedical and environmental detection at ppb sensitivity in biological samples without the requirement for any pre-treatment. This dual-pronged method combines the advantages of waste biodetoxification to produce a valuable chemical as well as a smart detection tool for environmental and biomedical detection.
- Watthaisong, Pratchaya,Pongpamorn, Pornkanok,Pimviriyakul, Panu,Maenpuen, Somchart,Ohmiya, Yoshihiro,Chaiyen, Pimchai
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supporting information
p. 13254 - 13258
(2019/08/16)
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- Assessment of the inhibitory effects of pyrethroids against human carboxylesterases
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Pyrethroids are broad-spectrum insecticides that widely used in many countries, while humans may be exposed to these toxins by drinking or eating pesticide-contaminated foods. This study aimed to investigate the inhibitory effects of six commonly used pyrethroids against two major human carboxylesterases (CES) including CES1 and CES2. Three optical probe substrates for CES1 (DME, BMBT and DMCB) and a fluorescent probe substrate for CES2 (DDAB) were used to characterize the inhibitory effects of these pyrethroids. The results demonstrated that most of the tested pyrethroids showed moderate to weak inhibitory effects against both CES1 and CES2, but deltamethrin displayed strong inhibition towards CES1. The IC50 values of deltamethrin against CES1-mediated BMBT, DME, and DMCB hydrolysis were determined as 1.58?μM, 2.39?μM, and 3.3?μM, respectively. Moreover, deltamethrin was cell membrane permeable and capable of inhibition endogenous CES1 in living cells. Further investigation revealed that deltamethrin inhibited CES1-mediated BMBT hydrolysis via competitive manner but noncompetitively inhibited DME or DMCB hydrolysis. The inhibition behaviors of deltamethrin against CES1 were also studied by molecular docking simulation. The results demonstrated that CES1 had at least two different ligand-binding sites, one was the DME site and another was the BMBT site which was identical to the binding site of deltamethrin. In summary, deltamethrin was a strong reversible inhibitor against CES1 and it could tightly bind on CES1 at the same ligand-binding site as BMBT. These findings are helpful for the deep understanding of the interactions between xenobiotics and CES1.
- Lei, Wei,Wang, Dan-Dan,Dou, Tong-Yi,Hou, Jie,Feng, Liang,Yin, Heng,Luo, Qun,Sun, Jie,Ge, Guang-Bo,Yang, Ling
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- Demethylating technology of 2-cyan-6-methoxy-benzothiazole, and D-fluorescein preparation method thereof
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The invention belongs to the field of chemical synthesis, and discloses a demethylating technology of 2-cyan-6-methoxy-benzothiazole. The technology comprises the following steps: 1, sequentially adding 2-cyan-6-methoxy-benzothiazole, metal iodide, mercaptan and acetonitrile to a reaction bottle, and stirring added materials for dissolving; 2, slowly adding an acetonitrile solution of trimethylchlorosilane to the above obtained system in a dropwise manner; 3, carrying out a heat insulation reaction after dropwise addition; and 4, adding an aqueous solution of an alkali in a dropwise manner after the reaction ends, adjusting the pH value to 7-12, adding an organic solvent, stirring the obtained solution, layering the stirred solution, backwashing the above obtained organic phase with water, drying the organic phase, and desolventizing the dried organic phase to obtain 2-cyan-6-hydroxy-benzothiazole. A reaction substrate and the above product can be well dissolved in the solvent selected in the invention, and the technology has the advantages of thorough reaction, high catalysis efficiency, reaction time shortening, mild reaction conditions, no high pressure, high temperature or other strict reaction conditions, high yield, and no generation of severely toxic substances. The final product is prepared from an intermediate through a one-kettle process without purification.
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Paragraph 0046; 0048
(2016/10/07)
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- Luciferin and derivatives as a DYRK selective scaffold for the design of protein kinase inhibitors
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D-Luciferin is widely used as a substrate in luciferase catalysed bioluminescence assays for in vitro studies. However, little is known about cross reactivity and potential interference of D-luciferin with other enzymes. We serendipitously found that firefly luciferin inhibited the CDK2/Cyclin A protein kinase. Inhibition profiling of D-luciferin over a 103-protein kinase panel showed significant inhibition of a small set of protein kinases, in particular the DYRK-family, but also other members of the CMGC-group, including ERK8 and CK2. Inhibition profiling on a 16-member focused library derived from D-luciferin confirms that D-luciferin represents a DYRK-selective chemotype of fragment-like molecular weight. Thus, observation of its inhibitory activity and the initial SAR information reported here promise to be useful for further design of protein kinase inhibitors with related scaffolds.
- Rothweiler, Ulli,Eriksson, Jonas,Stensen, Wenche,Leeson, Frederick,Engh, Richard A.,Svendsen, John S.
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p. 140 - 148
(2015/03/18)
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- Rapid and scalable assembly of firefly luciferase substrates
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Bioluminescence imaging with luciferase-luciferin pairs is a popular method for visualizing biological processes in vivo. Unfortunately, most luciferins are difficult to access and remain prohibitively expensive for some imaging applications. Here we report cost-effective and efficient syntheses of d-luciferin and 6′-aminoluciferin, two widely used bioluminescent substrates. Our approach employs inexpensive anilines and Appel's salt to generate the luciferin cores in a single pot. Additionally, the syntheses are scalable and can provide multi-gram quantities of both substrates. The streamlined production and improved accessibility of luciferin reagents will bolster in vivo imaging efforts. This journal is
- McCutcheon, David C.,Porterfield, William B.,Prescher, Jennifer A.
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p. 2117 - 2121
(2015/03/18)
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- PCL COMPOUNDS, COMPOSITIONS, AND METHODS OF USE THEREOF
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Disclosed herein are compounds useful for detecting oxidants in a living cell, in a multicellular organism, or in a cell-free sample. In particular, disclosed herein are bioluminescent reporter compounds, and more particularly, fluorinated peroxy-caged- luciferin (PCL) compounds, compositions comprising such compounds, methods of using such compounds and compositions, and processes for preparing such compounds. Also disclosed herein are kits and methods for detecting and measuring peroxynitrite, and optionally, additional oxidants in a living cell, in a multicellular organism, or in a cell-free sample.
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- Strategy for dual-analyte luciferin imaging: In vivo bioluminescence detection of hydrogen peroxide and caspase activity in a murine model of acute inflammation
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In vivo molecular imaging holds promise for understanding the underlying mechanisms of health, injury, aging, and disease, as it can detect distinct biochemical processes such as enzymatic activity, reactive small-molecule fluxes, or post-translational modifications. Current imaging techniques often detect only a single biochemical process, but, within whole organisms, multiple types of biochemical events contribute to physiological and pathological phenotypes. In this report, we present a general strategy for dual-analyte detection in living animals that employs in situ formation of firefly luciferin from two complementary caged precursors that can be unmasked by different biochemical processes. To establish this approach, we have developed Peroxy Caged Luciferin-2 (PCL-2), a H2O2-responsive boronic acid probe that releases 6-hydroxy-2-cyanobenzothiazole (HCBT) upon reacting with this reactive oxygen species, as well as a peptide-based probe, z-Ile-Glu-ThrAsp-d-Cys (IETDC), which releases d-cysteine in the presence of active caspase 8. Once released, HCBT and d-cysteine form firefly luciferin in situ, giving rise to a bioluminescent signal if and only if both chemical triggers proceed. This system thus constitutes an AND-type molecular logic gate that reports on the simultaneous presence of H2O2 and caspase 8 activity. Using these probes, chemoselective imaging of either H 2O2 or caspase 8 activity was performed in vitro and in vivo. Moreover, concomitant use of PCL-2 and IETDC in vivo establishes a concurrent increase in both H2O2 and caspase 8 activity during acute inflammation in living mice. Taken together, this method offers a potentially powerful new chemical tool for studying simultaneous oxidative stress and inflammation processes in living animals during injury, aging, and disease, as well as a versatile approach for concurrent monitoring of multiple analytes using luciferin-based bioluminescence imaging technologies.
- Van De Bittner, Genevieve C.,Bertozzi, Carolyn R.,Chang, Christopher J.
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p. 1783 - 1795
(2013/04/10)
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- A new synthesis of dehydroluciferin [2-(6′-hydroxy-2′- benzothiazolyl)-thiazole-4-carboxylic acid] from 1,4-benzoquinone
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A new synthesis of dehydroluciferin [2-(6′-hydroxy-2′- benzothiazolyl)-thiazole-4-carboxylic acid], the oxidative product of luciferin, has been realized starting from 1,4-benzoquinone. Reaction of this compound with l-cysteine ethyl ester, followed by an oxidation-cyclization step afforded 2-carbethoxy-6-hydroxybenzothiazole that was in situ hydrolyzed and decarboxylated to 6-hydroxybenzothiazole. The tert-butyl(dimethyl)silyl ether of this key intermediate was subjected to α-lithiation followed by formylation with DMF, and the resulting aldehyde condensed with l-cysteine ethyl ester. Dehydrogenation of the intermediate thiazolidine followed by deprotection afforded dehydroluciferin in 35% overall yield from 1,4-benzoquinone (69% from 6-hydroxybenzothiazole).
- Ciuffreda, Pierangela,Casati, Silvana,Meroni, Giuseppe,Santaniello, Enzo
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p. 5893 - 5897
(2013/07/27)
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- Expedient synthesis of electronically modified luciferins for bioluminescence imaging
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Bioluminescence imaging with luciferase enzymes requires access to light-emitting, small-molecule luciferins. Here, we describe a rapid method to synthesize d-luciferin, the substrate for firefly luciferase (Fluc), along with a novel set of electronically modified analogues. Our procedure utilizes a relatively rare, but synthetically useful dithiazolium reagent to generate heteroaromatic scaffolds in a divergent fashion. Two of the luciferin analogues produced with this approach emit light with Fluc in vitro and in live cells. Collectively, our work increases the number of substrates that can be used for bioluminescence imaging and provides a general strategy for synthesizing new collections of luciferins.
- McCutcheon, David C.,Paley, Miranda A.,Steinhardt, Rachel C.,Prescher, Jennifer A.
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p. 7604 - 7607
(2012/07/02)
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- Photoactivable bioluminescent probes for imaging luciferase activity
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A set of stable and efficient photoactivable bioluminescent probes for imaging luciferase activity has been developed, which displayed robust bioluminescent signals upon brief UV illumination in buffer, cells and living animals.
- Shao, Qing,Jiang, Tingting,Ren, Gang,Cheng, Zhen,Xing, Bengang
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supporting information; experimental part
p. 4028 - 4030
(2009/12/09)
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- A new synthesis of 2-cyano-6-hydroxybenzothiazole, the key intermediate of d-luciferin, starting from 1,4-benzoquinone
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2-Cyano-6-hydroxybenzothiazole is the key intermediate for the synthesis of d-luciferin, the natural substrate of firefly luciferases. A new synthesis of 2-cyano-6-hydroxybenzothiazole has been realized starting from the reaction of 1,4-benzoquinone with l-cysteine ethyl ester, followed by oxidation-cyclization of the intermediate ethyl (R)-2-amino-3-(2,5-dihydroxyphenylsulfanyl)propan-oate hydrochloride to 2-carbethoxy-6-hydroxybenzothiazole. A suitable protection of this intermediate and a conversion to the corresponding nitrile gave, after deprotection, 2-cyano-6-hydroxybenzothiazole (32% yield from 1,4-benzoquinone). This nitrile reacts with d-cysteine to afford d-luciferin at room temperature in nearly quantitative yield (90-95%).
- Meroni, Giuseppe,Ciana, Paolo,Maggi, Adriana,Santaniello, Enzo
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body text
p. 2682 - 2684
(2010/01/16)
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- A bioluminogenic substrate for in vivo imaging of β-lactamase activity
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Reporting live: A caged β-lactam-D-luciferin (Bluco) conjugate can image β-lactamase activity in vivo through a two-step enzymatic process. This novel bioluminogenic probe should facilitate the detection of β-lactamase and widen its applications as a reporter. (Chemical Equation Presented).
- Yao, Hequan,So, Min-Kyung,Rao, Jianghong
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p. 7031 - 7034
(2008/09/18)
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- Electrophilic aromatic substituted luciferins as bioluminescent probes for glutathione S-transferase assays
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New highly sensitive latent bioluminescent luciferin substrates were designed and synthesized for monitoring mammalian glutathione S-transferase (GST) and Schistosoma japonicum enzyme activities. The Royal Society of Chemistry 2006.
- Zhou, Wenhui,Shultz, John W.,Murphy, Nancy,Hawkins, Erika M.,Bernad, Laurent,Good, Troy,Moothart, Leonard,Frackman, Susan,Klaubert, Dieter H.,Bulleit, Robert F.,Wood, Keith V.
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p. 4620 - 4622
(2007/10/03)
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