Journal articles: 'Luciferin-luciferase bioluminescence'

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Simonyan, Hayk, Chansol Hurr, and Colin N. Young. "A synthetic luciferin improves in vivo bioluminescence imaging of gene expression in cardiovascular brain regions." Physiological Genomics 48, no. 10 (2016): 762–70. http://dx.doi.org/10.1152/physiolgenomics.00055.2016.

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Bioluminescence imaging is an effective tool for in vivo investigation of molecular processes. We have demonstrated the applicability of bioluminescence imaging to spatiotemporally monitor gene expression in cardioregulatory brain nuclei during the development of cardiovascular disease, via incorporation of firefly luciferase into living animals, combined with exogenous d-luciferin substrate administration. Nevertheless, d-luciferin uptake into the brain tissue is low, which decreases the sensitivity of bioluminescence detection, particularly when considering small changes in gene expression in tiny central areas. Here, we tested the hypothesis that a synthetic luciferin, cyclic alkylaminoluciferin (CycLuc1), would be superior to d-luciferin for in vivo bioluminescence imaging in cardiovascular brain regions. Male C57B1/6 mice underwent targeted delivery of an adenovirus encoding the luciferase gene downstream of the CMV promoter to the subfornical organ (SFO) or paraventricular nucleus of hypothalamus (PVN), two crucial cardioregulatory neural regions. While bioluminescent signals could be obtained following d-luciferin injection (150 mg/kg), CycLuc1 administration resulted in a three- to fourfold greater bioluminescent emission from the SFO and PVN, at 10- to 20-fold lower substrate concentrations (7.5–15 mg/kg). This CycLuc1-mediated enhancement in bioluminescent emission was evident early following substrate administration (i.e., 6–10 min) and persisted for up to 1 h. When the exposure time was reduced from 60 s to 1,500 ms, minimal signal in the PVN was detectable with d-luciferin, whereas bioluminescent images could be reliably captured with CycLuc1. These findings demonstrate that bioluminescent imaging with the synthetic luciferin CycLuc1 provides an improved physiological genomics tool to investigate molecular events in discrete cardioregulatory brain nuclei.

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Kotlobay, Alexey A., Maxim A. Dubinnyi, Konstantin V. Purtov, et al. "Bioluminescence chemistry of fireworm Odontosyllis." Proceedings of the National Academy of Sciences 116, no. 38 (2019): 18911–16. http://dx.doi.org/10.1073/pnas.1902095116.

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Marine polychaetes Odontosyllis undecimdonta, commonly known as fireworms, emit bright blue-green bioluminescence. Until the recent identification of the Odontosyllis luciferase enzyme, little progress had been made toward characterizing the key components of this bioluminescence system. Here we present the biomolecular mechanisms of enzymatic (leading to light emission) and nonenzymatic (dark) oxidation pathways of newly described O. undecimdonta luciferin. Spectral studies, including 1D and 2D NMR spectroscopy, mass spectrometry, and X-ray diffraction, of isolated substances allowed us to characterize the luciferin as an unusual tricyclic sulfur-containing heterocycle. Odontosyllis luciferin does not share structural similarity with any other known luciferins. The structures of the Odontosyllis bioluminescent system’s low molecular weight components have enabled us to propose chemical transformation pathways for the enzymatic and nonspecific oxidation of luciferin.

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Saito-Moriya, Ryohei, Jun Nakayama, Genta Kamiya, et al. "How to Select Firefly Luciferin Analogues for In Vivo Imaging." International Journal of Molecular Sciences 22, no. 4 (2021): 1848. http://dx.doi.org/10.3390/ijms22041848.

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Bioluminescence reactions are widely applied in optical in vivo imaging in the life science and medical fields. Such reactions produce light upon the oxidation of a luciferin (substrate) catalyzed by a luciferase (enzyme), and this bioluminescence enables the quantification of tumor cells and gene expression in animal models. Many researchers have developed single-color or multicolor bioluminescence systems based on artificial luciferin analogues and/or luciferase mutants, for application in vivo bioluminescence imaging (BLI). In the current review, we focus on the characteristics of firefly BLI technology and discuss the development of luciferin analogues for high-resolution in vivo BLI. In addition, we discuss the novel luciferin analogues TokeOni and seMpai, which show potential as high-sensitivity in vivo BLI reagents.

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Yull, Fiona E., Wei Han, E. Duco Jansen, et al. "Bioluminescent Detection of Endotoxin Effects on HIV-1 LTR-driven Transcription in Vivo." Journal of Histochemistry & Cytochemistry 51, no. 6 (2003): 741–49. http://dx.doi.org/10.1177/002215540305100605.

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We investigated the effects of Gram-negative bacterial lipopolysaccharide (LPS) on luciferase expression in transgenic reporter mice in which luciferase expression is driven by the nuclear factor κB (NF-κB)-dependent portion of the human immunodeficiency virus-1 (HIV-1) long terminal repeat (HIV-1 LTR). Using these mice, we dissected the sources of luciferase activity at the organ level by (a) assessing luciferase activity in organ homogenates, (b) bioluminescence imaging in vivo, and (c) bioluminescence imaging of individual organs ex vivo. Luciferin dosage was a critical determinant of the magnitude of photon emission from these reporter mice. Photon emission increased at doses from 0.5–6 mg of luciferin given by intraperitoneal (IP) injection. The differential between basal and LPS-induced bioluminescence was maximal at 3–6 mg of luciferin. Luciferase expression was highly inducible in lungs, liver, spleen, and kidneys after a single IP injection of LPS, as assessed by luciferase activity measurements in organ homogenates. Luciferase activity was also induced in the forebrain by treatment with IP LPS. In contrast, aerosolized LPS produced a response localized to the lungs as assessed by both bioluminescence and ex vivo luciferase assay measurements. These studies demonstrate the utility of luciferase reporter mice for determining organ-specific gene expression in response to local and systemic stimuli.

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Desjardins, Michel, and David Morse. "The polypeptide components of scintillons, the bioluminescence organelles of the dinoflagellate Gonyaulax polyedra." Biochemistry and Cell Biology 71, no. 3-4 (1993): 176–82. http://dx.doi.org/10.1139/o93-028.

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Scintillons, the bioluminescence organelles of Gonyaulax polyedra, were purified by isopycnic density gradient centrifugation until only low levels of contaminating chloroplasts and mitochondria were detected by fluorescence and electron microscopy. Purified scintillons catalyzed the luminescent reaction with kinetics identical to those observed during the bioluminescence flash in vivo. Polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate indicated that the organelles appeared to contain only two proteins. These proteins were identified as luciferase (135 kilodaltons) and luciferin-binding protein (75 kilodaltons) based on their size and their known functions in the bioluminescence reaction in vitro. The staining of luciferin-binding protein by Coomassie blue was 2.4 ± 0.3 (n = 19) times greater than the luciferase, suggesting that there are four binding protein monomers for every luciferase monomer. A model is proposed for the close packing of the two proteins inside the scintillons.Key words: luciferase, luciferin-binding protein, density gradient centrifugation, dinoflagellate.

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Jones, Krysten A., William B. Porterfield, Colin M. Rathbun, David C. McCutcheon, Miranda A. Paley, and Jennifer A. Prescher. "Orthogonal Luciferase–Luciferin Pairs for Bioluminescence Imaging." Journal of the American Chemical Society 139, no. 6 (2017): 2351–58. http://dx.doi.org/10.1021/jacs.6b11737.

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Viviani, Vadim R., Vanessa R. Bevilaqua, Daniel R. de Souza, Gabriel F. Pelentir, Michio Kakiuchi, and Takashi Hirano. "A Very Bright Far-Red Bioluminescence Emitting Combination Based on Engineered Railroad Worm Luciferase and 6′-Amino-Analogs for Bioimaging Purposes." International Journal of Molecular Sciences 22, no. 1 (2020): 303. http://dx.doi.org/10.3390/ijms22010303.

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Beetle luciferases produce bioluminescence (BL) colors ranging from green to red, having been extensively used for many bioanalytical purposes, including bioimaging of pathogen infections and metastasis proliferation in living animal models and cell culture. For bioimaging purposes in mammalian tissues, red bioluminescence is preferred, due to the lower self-absorption of light at longer wavelengths by hemoglobin, myoglobin and melanin. Red bioluminescence is naturally produced only by Phrixothrix hirtus railroad worm luciferase (PxRE), and by some engineered beetle luciferases. However, Far-Red (FR) and Near-Infrared (NIR) bioluminescence is best suited for bioimaging in mammalian tissues due to its higher penetrability. Although some FR and NIR emitting luciferin analogs have been already developed, they usually emit much lower bioluminescence activity when compared to the original luciferin-luciferases. Using site-directed mutagenesis of PxRE luciferase in combination with 6′-modified amino-luciferin analogs, we finally selected novel FR combinations displaying BL ranging from 636–655 nm. Among them, the combination of PxRE-R215K mutant with 6′-(1-pyrrolidinyl)luciferin proved to be the best combination, displaying the highest BL activity with a catalytic efficiency ~2.5 times higher than the combination with native firefly luciferin, producing the second most FR-shifted bioluminescence (650 nm), being several orders of magnitude brighter than commercial AkaLumine with firefly luciferase. Such combination also showed higher thermostability, slower BL decay time and better penetrability across bacterial cell membranes, resulting in ~3 times higher in vivo BL activity in bacterial cells than with firefly luciferin. Overall, this is the brightest FR emitting combination ever reported, and is very promising for bioimaging purposes in mammalian tissues.

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Endo, Mizuki, and Takeaki Ozawa. "Advanced Bioluminescence System for In Vivo Imaging with Brighter and Red-Shifted Light Emission." International Journal of Molecular Sciences 21, no. 18 (2020): 6538. http://dx.doi.org/10.3390/ijms21186538.

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In vivo bioluminescence imaging (BLI), which is based on luminescence emitted by the luciferase–luciferin reaction, has enabled continuous monitoring of various biochemical processes in living animals. Bright luminescence with a high signal-to-background ratio, ideally red or near-infrared light as the emission maximum, is necessary for in vivo animal experiments. Various attempts have been undertaken to achieve this goal, including genetic engineering of luciferase, chemical modulation of luciferin, and utilization of bioluminescence resonance energy transfer (BRET). In this review, we overview a recent advance in the development of a bioluminescence system for in vivo BLI. We also specifically examine the improvement in bioluminescence intensity by mutagenic or chemical modulation on several beetle and marine luciferase bioluminescence systems. We further describe that intramolecular BRET enhances luminescence emission, with recent attempts for the development of red-shifted bioluminescence system, showing great potency in in vivo BLI. Perspectives for future improvement of bioluminescence systems are discussed.

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Osipova, Z. M., A. S. Shcheglov, and I. V. Yampolsky. "A bioluminescent system of fungi: prospects for application in medical research." Alternatives to antibiotics, no. (1)2018 (March 4, 2018): 74–77. http://dx.doi.org/10.24075/brsmu.2018.004.

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Bioluminescence is chemical oxidation of a small luciferin molecule by air catalyzed by luciferase and accompanied by the emission of photons in the visible spectrum. This reaction is used in bioluminescent bioimaging, the method for the visualization of organism’s interior. Bioimaging is a popular tool used in medical research. However, it has an unfortunate drawback: it requires introduction of external luciferin to the system before every experiment. In this work we discuss a possibility of developing an autonomous luminescent system in eukaryotes based on the bioluminescent system of higher fungi.

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Shi, Ce, Michael P. Killoran, Mary P. Hall, et al. "5,5-Dialkylluciferins are thermal stable substrates for bioluminescence-based detection systems." PLOS ONE 15, no. 12 (2020): e0243747. http://dx.doi.org/10.1371/journal.pone.0243747.

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Firefly luciferase-based ATP detection assays are frequently used as a sensitive, cost-efficient method for monitoring hygiene in many industrial settings. Solutions of detection reagent, containing a mixture of a substrate and luciferase enzyme that produces photons in the presence of ATP, are relatively unstable and maintain only a limited shelf life even under refrigerated conditions. It is therefore common for the individual performing a hygiene test to manually prepare fresh reagent at the time of monitoring. To simplify sample processing, a liquid detection reagent with improved thermal stability is needed. The engineered firefly luciferase, Ultra-Glo™, fulfills one aspect of this need and has been valuable for hygiene monitoring because of its high resistance to chemical and thermal inactivation. However, solutions containing both Ultra-Glo™ luciferase and its substrate luciferin gradually lose the ability to effectively detect ATP over time. We demonstrate here that dehydroluciferin, a prevalent oxidative breakdown product of luciferin, is a potent inhibitor of Ultra-Glo™ luciferase and that its formation in the detection reagent is responsible for the decreased ability to detect ATP. We subsequently found that dialkylation at the 5-position of luciferin (e.g., 5,5-dimethylluciferin) prevents degradation to dehydroluciferin and improves substrate thermostability in solution. However, since 5,5-dialkylluciferins are poorly utilized by Ultra-Glo™ luciferase as substrates, we used structural optimization of the luciferin dialkyl modification and protein engineering of Ultra-Glo™ to develop a luciferase/luciferin pair that shows improved total reagent stability in solution at ambient temperature. The results of our studies outline a novel luciferase/luciferin system that could serve as foundations for the next generation of bioluminescence ATP detection assays with desirable reagent stability.

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Waldenmaier, Hans E., Anderson G. Oliveira, and Cassius V. Stevani. "Thoughts on the diversity of convergent evolution of bioluminescence on earth." International Journal of Astrobiology 11, no. 4 (2012): 335–43. http://dx.doi.org/10.1017/s1473550412000146.

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AbstractThe widespread independent evolution of analogous bioluminescent systems is one of the most impressive and diverse examples of convergent evolution on earth. There are roughly 30 extant bioluminescent systems that have evolved independently on Earth, with each system likely having unique enzymes responsible for catalysing the bioluminescent reaction. Bioluminescence is a chemical reaction involving a luciferin molecule and a luciferase or photoprotein that results in the emission of light. Some independent systems utilize the same luciferin, such as the use of tetrapyrrolic compounds by krill and dinoflagellates, and the wide use of coelenterazine by marine organisms, while the enzymes involved are unique. One common thread among all the different bioluminescent systems is the requirement of molecular oxygen. Bioluminescence is found in most forms of life, especially marine organisms.Bioluminescence in known to benefit the organism by: attraction, repulsion, communication, camouflage, and illumination. The marine ecosystem is significantly affected by bioluminescence, the only light found in the pelagic zone and below is from bioluminescent organisms.Transgenic bioluminescent organisms have revolutionized molecular research, medicine and the biotechnology industry. The use of bioluminescence in studying molecular pathways and disease allows for non-invasive and real-time analysis. Bioluminescence-based assays have been developed for several analytes by coupling luminescence to many enzyme-catalysed reactions.

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Tessler, Michael, Jean P. Gaffney, Jason M. Crawford, et al. "Luciferin production and luciferase transcription in the bioluminescent copepod Metridia lucens." PeerJ 6 (September 14, 2018): e5506. http://dx.doi.org/10.7717/peerj.5506.

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Bioluminescent copepods are often the most abundant marine zooplankton and play critical roles in oceanic food webs. Metridia copepods exhibit particularly bright bioluminescence, and the molecular basis of their light production has just recently begun to be explored. Here we add to this body of work by transcriptomically profiling Metridia lucens, a common species found in temperate, northern, and southern latitudes. In this previously molecularly-uncharacterized species, we find the typical luciferase paralog gene set found in Metridia. More surprisingly, we recover noteworthy putative luciferase sequences that had not been described from Metridia species, indicating that bioluminescence produced by these copepods may be more complex than previously known. This includes another copepod luciferase, as well as one from a shrimp. Furthermore, feeding experiments using mass spectrometry and 13C labelled L-tyrosine and L-phenylalanine firmly establish that M. lucens produces its own coelenterazine luciferin rather than acquiring it through diet. This coelenterazine synthesis has only been directly confirmed in one other copepod species.

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KrishnaMurthy, N. V., T. Sudhaharan, and A. Ram Reddy. "Dye induced quenching of firefly luciferase–luciferin bioluminescence." Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 68, no. 3 (2007): 851–59. http://dx.doi.org/10.1016/j.saa.2007.01.003.

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Feeney, Kevin A., Marrit Putker, Marco Brancaccio, and John S. O’Neill. "In-depth Characterization of Firefly Luciferase as a Reporter of Circadian Gene Expression in Mammalian Cells." Journal of Biological Rhythms 31, no. 6 (2016): 540–50. http://dx.doi.org/10.1177/0748730416668898.

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Firefly luciferase (Fluc) is frequently used to report circadian gene expression rhythms in mammalian cells and tissues. During longitudinal assays it is generally assumed that enzymatic substrates are in saturating excess, such that total bioluminescence is directly proportional to Fluc protein level. To test this assumption, we compared the enzyme kinetics of purified luciferase with its activity in mammalian cells. We found that Fluc activity in solution has a lower Michaelis constant (Km) for luciferin, lower temperature dependence, and lower catalytic half-life than Fluc in cells. In consequence, extracellular luciferin concentration significantly affects the apparent circadian amplitude and phase of the widely used PER2::LUC reporter in cultured fibroblasts, but not in SCN, and we suggest that this arises from differences in plasma membrane luciferin transporter activity. We found that at very high concentrations (>1 mM), luciferin lengthens circadian period, in both fibroblasts and organotypic SCN slices. We conclude that the amplitude and phase of circadian gene expression inferred from bioluminescence recordings should be treated with some caution, and we suggest that optimal luciferin concentration should be determined empirically for each luciferase reporter and cell type.

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Gregor, Carola, Jasmin K. Pape, Klaus C. Gwosch, Tanja Gilat, Steffen J. Sahl, and Stefan W. Hell. "Autonomous bioluminescence imaging of single mammalian cells with the bacterial bioluminescence system." Proceedings of the National Academy of Sciences 116, no. 52 (2019): 26491–96. http://dx.doi.org/10.1073/pnas.1913616116.

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Bioluminescence-based imaging of living cells has become an important tool in biological and medical research. However, many bioluminescence imaging applications are limited by the requirement of an externally provided luciferin substrate and the low bioluminescence signal which restricts the sensitivity and spatiotemporal resolution. The bacterial bioluminescence system is fully genetically encodable and hence produces autonomous bioluminescence without an external luciferin, but its brightness in cell types other than bacteria has, so far, not been sufficient for imaging single cells. We coexpressed codon-optimized forms of the bacterialluxCDABEandfrpgenes from multiple plasmids in different mammalian cell lines. Our approach produces high luminescence levels that are comparable to firefly luciferase, thus enabling autonomous bioluminescence microscopy of mammalian cells.

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Yeh, Hsien-Wei, Omran Karmach, Ao Ji, David Carter, Manuela M. Martins-Green, and Hui-wang Ai. "Red-shifted luciferase–luciferin pairs for enhanced bioluminescence imaging." Nature Methods 14, no. 10 (2017): 971–74. http://dx.doi.org/10.1038/nmeth.4400.

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Fajardo, Carlos, Marcos De Donato, Hectorina Rodulfo, et al. "New Perspectives Related to the Bioluminescent System in Dinoflagellates: Pyrocystis lunula, a Case Study." International Journal of Molecular Sciences 21, no. 5 (2020): 1784. http://dx.doi.org/10.3390/ijms21051784.

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Pyrocystis lunula is considered a model organism due to its bioluminescence capacity linked to circadian rhythms. The mechanisms underlying the bioluminescent phenomenon have been well characterized in dinoflagellates; however, there are still some aspects that remain an enigma. Such is the case of the presence and diversity of the luciferin-binding protein (LBP), as well as the synthesis process of luciferin. Here we carry out a review of the literature in relation to the molecular players responsible for bioluminescence in dinoflagellates, with particular interest in P. lunula. We also carried out a phylogenetic analysis of the conservation of protein sequence, structure and evolutionary pattern of these key players. The basic structure of the luciferase (LCF) is quite conserved among the sequences reported to date for dinoflagellate species, but not in the case of the LBP, which has proven to be more variable in terms of sequence and structure. In the case of luciferin, its synthesis has been shown to be complex process with more than one metabolic pathway involved. The glutathione S-transferase (GST) and the P630 or blue compound, seem to be involved in this process. In the same way, various hypotheses regarding the role of bioluminescence in dinoflagellates are exposed.

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Miceli, Matteo, Silvana Casati, Pietro Allevi, et al. "A New Ultrasensitive Bioluminescence-Based Method for Assaying Monoacylglycerol Lipase." International Journal of Molecular Sciences 22, no. 11 (2021): 6148. http://dx.doi.org/10.3390/ijms22116148.

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A novel bioluminescent Monoacylglycerol lipase (MAGL) substrate 6-O-arachidonoylluciferin, a D-luciferin derivative, was synthesized, physico-chemically characterized, and used as highly sensitive substrate for MAGL in an assay developed for this purpose. We present here a new method based on the enzymatic cleavage of arachidonic acid with luciferin release using human Monoacylglycerol lipase (hMAGL) followed by its reaction with a chimeric luciferase, PLG2, to produce bioluminescence. Enzymatic cleavage of the new substrate by MAGL was demonstrated, and kinetic constants Km and Vmax were determined. 6-O-arachidonoylluciferin has proved to be a highly sensitive substrate for MAGL. The bioluminescence assay (LOD 90 pM, LOQ 300 pM) is much more sensitive and should suffer fewer biological interferences in cells lysate applications than typical fluorometric methods. The assay was validated for the identification and characterization of MAGL modulators using the well-known MAGL inhibitor JZL184. The use of PLG2 displaying distinct bioluminescence color and kinetics may offer a highly desirable opportunity to extend the range of applications to cell-based assays.

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Ikeda, Yuma, Tsuyoshi Saitoh, Kazuki Niwa, et al. "An allylated firefly luciferin analogue with luciferase specific response in living cells." Chemical Communications 54, no. 14 (2018): 1774–77. http://dx.doi.org/10.1039/c7cc09720d.

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Kotlobay, A. A., Z. M. Kaskova, and I. V. Yampolsky. "Palette of Luciferases: Natural Biotools for New Applications in Biomedicine." Acta Naturae 12, no. 2 (2020): 15–27. http://dx.doi.org/10.32607/actanaturae.10967.

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Optoanalytical methods based on using genetically encoded bioluminescent enzymes,luciferases, allow one to obtain highly sensitive signals, are non-invasive, and require no external irradiation. Bioluminescence is based on the chemical reaction of oxidation of a low-molecular-weight substrate (luciferin) by atmospheric oxygen, which is catalyzed by an enzyme (luciferase). Relaxation of the luciferin oxidation product from its excited state is accompanied by a release of a quantum of light, which can be detected as an analytical signal.The ability to express luciferase genes in various heterological systems and high quantum yields of luminescence reactions have made these tools rather popular in biology and medicine. Amongseveral naturally available luciferases, a few have been found to be useful for practicalapplication. Luciferase size, the wavelength of its luminescence maximum, enzyme thermostability, optimal pH of the reaction, and the need for cofactors areparameters that may differ for luciferases from different groups of organisms, and this fact directly affects the choice of the application area for each enzyme. It is quite important to overview the whole range of currently available luciferases based ontheir biochemical properties before choosing one bioluminescent probe suitable for a specific application.

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Kotlobay, A. A., Z. M. Kaskova, and I. V. Yampolsky. "Palette of Luciferases: Natural Biotools for New Applications in Biomedicine." Acta Naturae 12, no. 2 (2020): 15–27. http://dx.doi.org/10.32607/actanaturae.11152.

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Optoanalytical methods based on using genetically encoded bioluminescent enzymes,luciferases, allow one to obtain highly sensitive signals, are non-invasive, and require no external irradiation. Bioluminescence is based on the chemical reaction of oxidation of a low-molecular-weight substrate (luciferin) by atmospheric oxygen, which is catalyzed by an enzyme (luciferase). Relaxation of the luciferin oxidation product from its excited state is accompanied by a release of a quantum of light, which can be detected as an analytical signal.The ability to express luciferase genes in various heterological systems and high quantum yields of luminescence reactions have made these tools rather popular in biology and medicine. Amongseveral naturally available luciferases, a few have been found to be useful for practicalapplication. Luciferase size, the wavelength of its luminescence maximum, enzyme thermostability, optimal pH of the reaction, and the need for cofactors areparameters that may differ for luciferases from different groups of organisms, and this fact directly affects the choice of the application area for each enzyme. It is quite important to overview the whole range of currently available luciferases based ontheir biochemical properties before choosing one bioluminescent probe suitable for a specific application.

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Shao, C. Y., C. J. Howe, A. J. R. Porter, and L. A. Glover. "Novel Cyanobacterial Biosensor for Detection of Herbicides." Applied and Environmental Microbiology 68, no. 10 (2002): 5026–33. http://dx.doi.org/10.1128/aem.68.10.5026-5033.2002.

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ABSTRACT The aim of this work was to generate a cyanobacterial biosensor that could be used to detect herbicides and other environmental pollutants. A representative freshwater cyanobacterium, Synechocystis sp. strain PCC6803, was chromosomally marked with the luciferase gene luc (from the firefly Photinus pyralis) to create a novel bioluminescent cyanobacterial strain. Successful expression of the luc gene during growth of Synechocystis sp. strain PCC6803 cultures was characterized by measuring optical density and bioluminescence. Bioluminescence was optimized with regard to uptake of the luciferase substrate, luciferin, and the physiology of the cyanobacterium. Bioassays demonstrated that a novel luminescent cyanobacterial biosensor has been developed which responded to a range of compounds including different herbicide types and other toxins. This biosensor is expected to provide new opportunities for the rapid screening of environmental samples or for the investigation of potential environmental damage.

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Minekawa, Takayuki, Hiroshi Ohkuma, Katsushi Abe, Hiroaki Maekawa, and Hidetoshi Arakawa. "Practical application of bioluminescence enzyme immunoassay using enhancer for firefly luciferin-luciferase bioluminescence." Luminescence 26, no. 3 (2010): 167–71. http://dx.doi.org/10.1002/bio.1200.

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Vongsangnak, Wanwipa, Pramote Chumnanpuen, and Ajaraporn Sriboonlert. "Transcriptome analysis reveals candidate genes involved in luciferin metabolism inLuciola aquatilis(Coleoptera: Lampyridae)." PeerJ 4 (October 4, 2016): e2534. http://dx.doi.org/10.7717/peerj.2534.

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Bioluminescence, which living organisms such as fireflies emit light, has been studied extensively for over half a century. This intriguing reaction, having its origins in nature where glowing insects can signal things such as attraction or defense, is now widely used in biotechnology with applications of bioluminescence and chemiluminescence. Luciferase, a key enzyme in this reaction, has been well characterized; however, the enzymes involved in the biosynthetic pathway of its substrate, luciferin, remains unsolved at present. To elucidate the luciferin metabolism, we performed ade novotranscriptome analysis using larvae of the firefly species,Luciola aquatilis. Here, a comparative analysis is performed with the model coleopteran insectTribolium casteneumto elucidate the metabolic pathways inL. aquatilis. Based on a template luciferin biosynthetic pathway, combined with a range of protein and pathway databases, and various prediction tools for functional annotation, the candidate genes, enzymes, and biochemical reactions involved in luciferin metabolism are proposed forL. aquatilis. The candidate gene expression is validated in the adultL. aquatilisusing reverse transcription PCR (RT-PCR). This study provides useful information on the bio-production of luciferin in the firefly and will benefit to future applications of the valuable firefly bioluminescence system.

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Jawhara, Samir, and Serge Mordon. "In Vivo Imaging of Bioluminescent Escherichia coli in a Cutaneous Wound Infection Model for Evaluation of an Antibiotic Therapy." Antimicrobial Agents and Chemotherapy 48, no. 9 (2004): 3436–41. http://dx.doi.org/10.1128/aac.48.9.3436-3441.2004.

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ABSTRACT A rapid, continuous method for noninvasively monitoring the effectiveness of several antibacterial agents in real time by using a model of wound infection was developed. This study was divided into three steps: (i) construction of a plasmid to transform Escherichia coli into a bioluminescent variant, (ii) study of the bioluminescent E. coli in vitro as a function of temperature and pH, and (iii) determination of the MIC and the minimal bactericidal concentration of sulfamethoxazole-trimethoprim (SMX-TMP). Finally, the efficacy of SMX-TMP was monitored in vivo in a cutaneous wound model (hairless rat) infected with this bioluminescent bacterium by using a bioluminescence imaging system. E. coli was transformed by electroporation with a shuttle vector (pRB474) containing the firefly (Photinus pyralis) luciferase gene, resulting in a bioluminescent phenotype. It was found that pH 5.0 was optimal for incorporation of the susbstrate d-luciferin for the luciferase reaction. In vitro, when the agar dilution method, standard turbidity assays, and the bioluminescence imaging system were used, E. coli(pRB474) proved to be susceptible to SMX-TMP. In vivo, at 4 h, SMX-TMP treatment was already efficient compared to no treatment (P = 0.034). At 48 h, no bioluminescence was detected in the wound, demonstrating the susceptibility of E. coli to SMX-TMP. In conclusion, this study points out the advantage of using bioluminescence imaging to evaluate the effects of antibiotics for the treatment of acute infections in vivo in a nondestructive and noninvasive manner.

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Kotlobay, Alexey A., Karen S. Sarkisyan, Yuliana A. Mokrushina, et al. "Genetically encodable bioluminescent system from fungi." Proceedings of the National Academy of Sciences 115, no. 50 (2018): 12728–32. http://dx.doi.org/10.1073/pnas.1803615115.

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Bioluminescence is found across the entire tree of life, conferring a spectacular set of visually oriented functions from attracting mates to scaring off predators. Half a dozen different luciferins, molecules that emit light when enzymatically oxidized, are known. However, just one biochemical pathway for luciferin biosynthesis has been described in full, which is found only in bacteria. Here, we report identification of the fungal luciferase and three other key enzymes that together form the biosynthetic cycle of the fungal luciferin from caffeic acid, a simple and widespread metabolite. Introduction of the identified genes into the genome of the yeastPichia pastorisalong with caffeic acid biosynthesis genes resulted in a strain that is autoluminescent in standard media. We analyzed evolution of the enzymes of the luciferin biosynthesis cycle and found that fungal bioluminescence emerged through a series of events that included two independent gene duplications. The retention of the duplicated enzymes of the luciferin pathway in nonluminescent fungi shows that the gene duplication was followed by functional sequence divergence of enzymes of at least one gene in the biosynthetic pathway and suggests that the evolution of fungal bioluminescence proceeded through several closely related stepping stone nonluminescent biochemical reactions with adaptive roles. The availability of a complete eukaryotic luciferin biosynthesis pathway provides several applications in biomedicine and bioengineering.

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Oba, Y., K. Konishi, D. Yano, H. Shibata, D. Kato, and T. Shirai. "Resurrecting the ancient glow of the fireflies." Science Advances 6, no. 49 (2020): eabc5705. http://dx.doi.org/10.1126/sciadv.abc5705.

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The color of firefly bioluminescence is determined by the structure of luciferase. Firefly luciferase genes have been isolated from more than 30 species, producing light ranging in color from green to orange-yellow. Here, we reconstructed seven ancestral firefly luciferase genes, characterized the enzymatic properties of the recombinant proteins, and determined the crystal structures of the gene from ancestral Lampyridae. Results showed that the synthetic luciferase for the last common firefly ancestor exhibited green light caused by a spatial constraint on the luciferin molecule in enzyme, while fatty acyl-CoA synthetic activity, an original function of firefly luciferase, was diminished in exchange. All known firefly species are bioluminescent in the larvae, with a common ancestor arising approximately 100 million years ago. Combined, our findings propose that, within the mid-Cretaceous forest, the common ancestor of fireflies evolved green light luciferase via trade-off of the original function, which was likely aposematic warning display against nocturnal predation.

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Li, Shufeng, Zhiyang Ruan, Hang Zhang, and Haiwei Xu. "Recent achievements of bioluminescence imaging based on firefly luciferin-luciferase system." European Journal of Medicinal Chemistry 211 (February 2021): 113111. http://dx.doi.org/10.1016/j.ejmech.2020.113111.

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Niwa, Kazuki, Yoshihiro Nakajima, and Yoshihiro Ohmiya. "Applications of luciferin biosynthesis: Bioluminescence assays for l-cysteine and luciferase." Analytical Biochemistry 396, no. 2 (2010): 316–18. http://dx.doi.org/10.1016/j.ab.2009.09.014.

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Wang, Anni, Xuewei Li, Yong Ju, Dongying Chen, and Jianzhong Lu. "Bioluminescence imaging of carbon monoxide in living cells based on a selective deiodination reaction." Analyst 145, no. 2 (2020): 550–56. http://dx.doi.org/10.1039/c9an02107h.

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Modification of a heavy iodine atom for d-Luciferin was explored as a “turn-on” transduction scheme for CO detection. This new probe could image exogenous and endogenous CO in the luciferase-transfected cancer cells.

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Delroisse, Jérôme, Esther Ullrich-Lüter, Stefanie Blaue, et al. "A puzzling homology: a brittle star using a putative cnidarian-type luciferase for bioluminescence." Open Biology 7, no. 4 (2017): 160300. http://dx.doi.org/10.1098/rsob.160300.

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Bioluminescence relies on the oxidation of a luciferin substrate catalysed by a luciferase enzyme. Luciferins and luciferases are generic terms used to describe a large variety of substrates and enzymes. Whereas luciferins can be shared by phylogenetically distant organisms which feed on organisms producing them, luciferases have been thought to be lineage-specific enzymes. Numerous light emission systems would then have co-emerged independently along the tree of life resulting in a plethora of non-homologous luciferases. Here, we identify for the first time a candidate luciferase of a luminous echinoderm, the ophiuroid Amphiura filiformis . Phylogenomic analyses identified the brittle star predicted luciferase as homologous to the luciferase of the sea pansy Renilla (Cnidaria), contradicting with the traditional viewpoint according to which luciferases would generally be of convergent origins. The similarity between the Renilla and Amphiura luciferases allowed us to detect the latter using anti- Renilla luciferase antibodies. Luciferase expression was specifically localized in the spines which were demonstrated to be the bioluminescent organs in vivo . However, enzymes homologous to the Renilla luciferase but unable to trigger light emission were also identified in non-luminous echinoderms and metazoans. Our findings strongly indicate that those enzymes, belonging to the haloalkane dehalogenase family, might then have been convergently co-opted into luciferases in cnidarians and echinoderms. In these two benthic suspension-feeding species, similar ecological pressures would constitute strong selective forces for the functional shift of these enzymes and the emergence of bioluminescence.

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Zambito, Giorgia, Natasa Gaspar, Yanto Ridwan, et al. "Evaluating Brightness and Spectral Properties of Click Beetle and Firefly Luciferases Using Luciferin Analogues: Identification of Preferred Pairings of Luciferase and Substrate for In Vivo Bioluminescence Imaging." Molecular Imaging and Biology 22, no. 6 (2020): 1523–31. http://dx.doi.org/10.1007/s11307-020-01523-7.

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Abstract Purpose Currently, a variety of red and green beetle luciferase variants are available for bioluminescence imaging (BLI). In addition, new luciferin analogues providing longer wavelength luminescence have been developed that show promise for improved deep tissue imaging. However, a detailed assessment of these analogues (e.g., Akalumine-HCl, CycLuc1, and amino naphthyl luciferin (NH2-NpLH2)) combined with state of the art luciferases has not been performed. The aim of this study was to evaluate for the first time the in vivo brightness and spectral characteristics of firefly (Luc2), click beetle green (CBG99), click beetle red 2 (CBR2), and Akaluc luciferases when paired with different d-luciferin (d-LH2) analogues in vivo. Procedures Transduced human embryonic kidney (HEK 293T) cells expressing individual luciferases were analyzed both in vitro and in mice (via subcutaneous injection). Following introduction of the luciferins to cells or animals, the resulting bioluminescence signal and photon emission spectrum were acquired using a sensitive charge-coupled device (CCD) camera equipped with a series of band pass filters and spectral unmixing software. Results Our in vivo analysis resulted in four primary findings: (1) the best substrate for Luc2, CBG99, and CBR2 in terms of signal strength was d-luciferin; (2) the spectra for Luc2 and CBR2 were shifted to a longer wavelength when Akalumine-HCl was the substrate; (3) CBR2 gave the brightest signal with the near-infrared substrate, NH2-NpLH2; and (4) Akaluc was brighter when paired with either CycLuc1 or Akalumine-HCl when paired with d-LH2. Conclusion We believe that the experimental results described here should provide valuable guidance to end users for choosing the correct luciferin/luciferase pairs for a variety of BLI applications.

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Fang, Zhongjian, Houchao Xu, Xiangjun Ji, et al. "Study of the Dynamic Uptake of Free Drug and Nanostructures for Drug Delivery Based on Bioluminescence Measurements." Journal of Nanomaterials 2017 (2017): 1–12. http://dx.doi.org/10.1155/2017/8542806.

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The past two decades have witnessed the great growth of the development of novel drug carriers. However, the releasing dynamics of drug from drug carriers in vivo and the interactions between cells and drug carriers remain unclear. In this paper, liposomes were prepared to encapsulate D-luciferin, which was the substrate of luciferase and served as a model drug. Based on the theoretical calculation of active loading, methods of preparation for liposomes were optimized. Only when D-luciferin was released from liposomes or taken in by the cells could bioluminescence be produced under the catalysis of luciferase. Models of multicellular tumor spheroid (MCTS) were built with 4T1-luc cells that expressed luciferase stably. The kinetic processes of uptake and distribution of free drugs and liposomal drugs were determined with models of cell suspension, monolayer cells, MCTS, and tumor-bearing nude mice. The technology platform has been demonstrated to be effective for the study of the distribution and kinetic profiles of various liposomes as drug delivery systems.

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Hara, Kiyotaka Y., and Hideo Mori. "An Efficient Method for Quantitative Determination of Cellular ATP Synthetic Activity." Journal of Biomolecular Screening 11, no. 3 (2006): 310–17. http://dx.doi.org/10.1177/1087057105285112.

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The authors have developed an efficient method to measure cellular activity of ATP synthesis. Although ATP is a major energy source of biological reactions, it has been difficult tomeasure cellular ATP synthetic activity quantitatively. In this report, bioluminescence from the luciferin-luciferase reactionwas used for the quantitativemeasurement. Under the used condition, bioluminescence from standard ATP solution showed no attenuation within several minutes, and the intensity corresponded proportionally to ATP concentrations of the standards. To measure dynamic cellular ATP synthetic activity, combination of osmotic shock and detergent treatmentwas used tomake Escherichia coli cells permeable. ATPwas discharged from permeable cells and reacted with externally added luciferase. Because permeable cells used glucose to synthesize and accumulate ATP without further growth, intensity of bioluminescence was increasing during the cellular consumption of glucose. Cellular ATP biosynthetic activitywas calculated formthe slope of linearly increasing bioluminescence. This permeable cell assay could be applied to high-throughput measuring for dynamic cellular activity of glycolytic ATP synthesis.

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Yoshida, Masa-aki, Junichi Imoto, Yuri Kawai, et al. "Genomic and Transcriptomic Analyses of Bioluminescence Genes in the Enope Squid Watasenia scintillans." Marine Biotechnology 22, no. 6 (2020): 760–71. http://dx.doi.org/10.1007/s10126-020-10001-8.

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AbstractWatasenia scintillans, a sparkling enope squid, has bioluminescence organs to illuminate its body with its own luciferase activity. To clarify the molecular mechanism underlying its scintillation, we analysed high-throughput sequencing data acquired previously and obtained draft genome sequences accomplished with comparative genomic data among the cephalopods. The genome mapped by transcriptome data showed that (1) RNA editing contributed to transcriptome variation of lineage specific genes, such as W. scintillans luciferase, and (2) two types of luciferase enzymes were characterized with reasonable 3D models docked to a luciferin molecule. We report two different types of luciferase in one organism and possibly related to variety of colour types in the W. scintillans fluorescent organs.

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Bessho-Uehara, Manabu, Naoyuki Yamamoto, Shuji Shigenobu, Hitoshi Mori, Keiko Kuwata, and Yuichi Oba. "Kleptoprotein bioluminescence: Parapriacanthus fish obtain luciferase from ostracod prey." Science Advances 6, no. 2 (2020): eaax4942. http://dx.doi.org/10.1126/sciadv.aax4942.

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Through their diet, animals can obtain substances essential for imparting special characteristics, such as toxins in monarch butterflies and luminescent substances in jellyfishes. These substances are typically small molecules because they are less likely to be digested and may be hard for the consumer to biosynthesize. Here, we report that Parapriacanthus ransonneti, a bioluminescent fish, obtains not only its luciferin but also its luciferase enzyme from bioluminescent ostracod prey. The enzyme purified from the fish’s light organs was identical to the luciferase of Cypridina noctiluca, a bioluminescent ostracod that they feed upon. Experiments where fish were fed with a related ostracod, Vargula hilgendorfii, demonstrated the specific uptake of the luciferase to the fish’s light organs. This “kleptoprotein” system allows an organism to use novel functional proteins that are not encoded in its genome and provides an evolutionary alternative to DNA-based molecular evolution.

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Pajor, Katarzyna, Daniel Sypniewski, and Ilona Bednarek. "Bioluminescence as a tool in molecular biology." Postępy Higieny i Medycyny Doświadczalnej 71 (December 12, 2017): 0. http://dx.doi.org/10.5604/01.3001.0010.7011.

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Bioluminescence has been studied for many years by scientists. There are numerous mechanisms of that phenomenon; among them bacterial bioluminescence is the most frequently found in nature. This type of bioluminescence is determined by the appearance of lux operon, which encodes all elements necessary to produce light emission and it does not require any additional substrates supply. Another commonly found example of bioluminescence mechanism is performed by Photinus pyralis. Luciferase of P. pyralis named FLuc requires D-luciferin as a substrate. Bioluminescence is also characteristic for many deep-sea organisms. Most of them are based on oxidation reaction of coelenterazine to coelenteramide mediated by RLuc or GLuc luciferases. Due to the variety of bioluminescence mechanisms in nature, it has become possible to apply them in many sensitive methods that can be used in molecular biology and medicine. The most significant application of bioluminescence is BLI (bioluminescence imaging). This method is cheap and nontoxic which allows both in vitro and in vivo imaging. BLI applications include, e.g. protein-protein interactions, stem cells labeling, tracking of viral, bacterial, fungal and parasitical infections, and carcinogenesis analyses. Bioluminescence has also been used in the creation of modified cell systems capable of light emission in response to certain analytes and thus very sensitive biosensors have been generated. Other important areas of bioluminescence application are immunoassays, ATP assays, and BART analysis (bioluminescent assay in Real-Time) – a very sensitive technique which allows scientists to estimate nucleic acids amplification.

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Nakayama, Jun, Ryohei Saito, Yusuke Hayashi, et al. "High Sensitivity In Vivo Imaging of Cancer Metastasis Using a Near-Infrared Luciferin Analogue seMpai." International Journal of Molecular Sciences 21, no. 21 (2020): 7896. http://dx.doi.org/10.3390/ijms21217896.

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Bioluminescence imaging (BLI) is useful to monitor cell movement and gene expression in live animals. However, D-luciferin has a short wavelength (560 nm) which is absorbed by tissues and the use of near-infrared (NIR) luciferin analogues enable high sensitivity in vivo BLI. The AkaLumine-AkaLuc BLI system (Aka-BLI) can detect resolution at the single-cell level; however, it has a clear hepatic background signal. Here, to enable the highly sensitive detection of bioluminescence from the surrounding liver tissues, we focused on seMpai (C15H16N3O2S) which has been synthesized as a luciferin analogue and has high luminescent abilities as same as AkaLumine. We demonstrated that seMpai BLI could detect micro-signals near the liver without any background signal. The solution of seMpai was neutral; therefore, seMpai imaging did not cause any adverse effect in mice. seMpai enabled a highly sensitive in vivo BLI as compared to previous techniques. Our findings suggest that the development of a novel mutated luciferase against seMpai may enable a highly sensitive BLI at the single-cell level without any background signal. Novel seMpai BLI system can be used for in vivo imaging in the fields of life sciences and medicine.

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Loimaranta, Vuokko, Jorma Tenovuo, Leeni Koivisto, and Matti Karp. "Generation of Bioluminescent Streptococcus mutans and Its Usage in Rapid Analysis of the Efficacy of Antimicrobial Compounds." Antimicrobial Agents and Chemotherapy 42, no. 8 (1998): 1906–10. http://dx.doi.org/10.1128/aac.42.8.1906.

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ABSTRACT The oral bacterium Streptococcus mutans was transformed by electroporation with a shuttle vector (pCSS945) containing insect luciferase gene from a click beetle (Pyrophorus plagiophthalamus) resulting in a bioluminescent phenotype. ThisS. mutans strain was used in experiments in which light emission was used as a rapid and, compared to conventional CFU counting, more convenient means of estimating the effects of various antimicrobial treatments. The basic parameters affecting in vivo light production by the strain were studied. It was found that pH 6.0 was optimal for incorporation of the substrate d-luciferin for the luciferase reaction. The optimum concentration ofd-luciferin was approximately 150 μM at room temperature. Under optimum conditions the light emission in vivo increased rapidly to a constant level and thereafter had a decay of 0.6%/min when logarithmic-growth-phase cells were used. The light emission closely paralleled the numbers of CFU, giving a detectable signal from 30,000 cells and having a dynamic measurement range over 4 log CFU/relative light unit. The cells were treated with various antimicrobial agents, and the emitted bioluminescence was measured. With the bioluminescent measurements, the results were obtained within hours compared to the days required for agar plates, and also, the kinetics of the antibacterial actions could be followed. Thus, the light emission was found to be a reliable, sensitive, and real-time indicator of the bacteriostatic actions of the antimicrobial agents tested.

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ZHENG, YING, QIAOYA LIN, HONGLIN JIN, JUAN CHEN, and ZHIHONG ZHANG. "VISUALIZATION OF HEAD AND NECK CANCER MODELS WITH A TRIPLE FUSION REPORTER GENE." Journal of Innovative Optical Health Sciences 05, no. 04 (2012): 1250028. http://dx.doi.org/10.1142/s1793545812500289.

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The development of experimental animal models for head and neck tumors generally rely on the bioluminescence imaging to achieve the dynamic monitoring of the tumor growth and metastasis due to the complicated anatomical structures. Since the bioluminescence imaging is largely affected by the intracellular luciferase expression level and external D-luciferin concentrations, its imaging accuracy requires further confirmation. Here, a new triple fusion reporter gene, which consists of a herpes simplex virus type 1 thymidine kinase (TK) gene for radioactive imaging, a far-red fluorescent protein (mLumin) gene for fluorescent imaging, and a firefly luciferase gene for bioluminescence imaging, was introduced for in vivo observation of the head and neck tumors through multi-modality imaging. Results show that fluorescence and bioluminescence signals from mLumin and luciferase, respectively, were clearly observed in tumor cells, and TK could activate suicide pathway of the cells in the presence of nucleotide analog-ganciclovir (GCV), demonstrating the effectiveness of individual functions of each gene. Moreover, subcutaneous and metastasis animal models for head and neck tumors using the fusion reporter gene-expressing cell lines were established, allowing multi-modality imaging in vivo. Together, the established tumor models of head and neck cancer based on the newly developed triple fusion reporter gene are ideal for monitoring tumor growth, assessing the drug therapeutic efficacy and verifying the effectiveness of new treatments.

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Lapointe, Mathieu, and David Morse. "Reassessing the role of a 3′-UTR-binding translational inhibitor in regulation of circadian bioluminescence rhythm in the dinoflagellate Gonyaulax." Biological Chemistry 389, no. 1 (2008): 13–19. http://dx.doi.org/10.1515/bc.2008.003.

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Abstract The nightly bioluminescence of the dinoflagellate Gonyaulax is a circadian rhythm caused by the presence in cells of specialized bioluminescent organelles, termed scintillons, containing the reaction catalyst luciferase, the substrate luciferin and a luciferin-binding protein (LBP). LBP levels increase at the start of the night phase because of increased protein synthesis rates in vivo, and this regulation has been ascribed to circadian binding of an inhibitory protein factor binding to the 3′ untranslated region (UTR) of lbp mRNA at times when LBP is not normally synthesized. To purify and characterize the binding factor, the electrophoretic mobility shift assays and UV crosslinking experiments used to first characterize the factor were repeated. However, neither these protocols nor binding to biotinylated RNA probes confirmed the presence of a specific circadian RNA-binding protein. Furthermore, neither RNA probe screening of a cDNA library expressed in bacteria nor three-hybrid assays in yeast were successful in isolating a cDNA encoding a protein able to bind specifically to the lbp 3′UTR. Taken together, these results suggest that alternative mechanisms for regulating lbp translation should now be examined.

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Heffern, Marie C., Hyo Min Park, Ho Yu Au-Yeung, et al. "In vivo bioluminescence imaging reveals copper deficiency in a murine model of nonalcoholic fatty liver disease." Proceedings of the National Academy of Sciences 113, no. 50 (2016): 14219–24. http://dx.doi.org/10.1073/pnas.1613628113.

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Copper is a required metal nutrient for life, but global or local alterations in its homeostasis are linked to diseases spanning genetic and metabolic disorders to cancer and neurodegeneration. Technologies that enable longitudinal in vivo monitoring of dynamic copper pools can help meet the need to study the complex interplay between copper status, health, and disease in the same living organism over time. Here, we present the synthesis, characterization, and in vivo imaging applications of Copper-Caged Luciferin-1 (CCL-1), a bioluminescent reporter for tissue-specific copper visualization in living animals. CCL-1 uses a selective copper(I)-dependent oxidative cleavage reaction to released-luciferin for subsequent bioluminescent reaction with firefly luciferase. The probe can detect physiological changes in labile Cu+levels in live cells and mice under situations of copper deficiency or overload. Application of CCL-1 to mice with liver-specific luciferase expression in a diet-induced model of nonalcoholic fatty liver disease reveals onset of hepatic copper deficiency and altered expression levels of central copper trafficking proteins that accompany symptoms of glucose intolerance and weight gain. The data connect copper dysregulation to metabolic liver disease and provide a starting point for expanding the toolbox of reactivity-based chemical reporters for cell- and tissue-specific in vivo imaging.

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Gandelman, O. A., L. Yu Brovko, N. N. Ugarova, A. Yu Chikishev, and A. P. Shkurimov. "Oxyluciferin fluorescence is a model of native bioluminescence in the firefly luciferin—luciferase system." Journal of Photochemistry and Photobiology B: Biology 19, no. 3 (1993): 187–91. http://dx.doi.org/10.1016/1011-1344(93)87083-y.

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Mizuno, Gaku, Daichi Yano, José Paitio, Hiromitsu Endo, and Yuichi Oba. "Etmopterus lantern sharks use coelenterazine as the substrate for their luciferin-luciferase bioluminescence system." Biochemical and Biophysical Research Communications 577 (November 2021): 139–45. http://dx.doi.org/10.1016/j.bbrc.2021.09.007.

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Khalil, Ashraf A., Mark J. Jameson, William C. Broaddus, et al. "Subcutaneous Administration of D-Luciferin is an Effective Alternative to Intraperitoneal Injection in Bioluminescence Imaging of Xenograft Tumors in Nude Mice." ISRN Molecular Imaging 2013 (December 18, 2013): 1–7. http://dx.doi.org/10.1155/2013/689279.

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Currently, intraperitoneal (IP) injection of D-luciferin is the preferred method of providing substrate for bioluminescence imaging (BLI); however it has a failure rate of 3–10% due to accidental intestinal injection. The present study evaluates the quality of BLI after subcutaneous (SC) injection of D-luciferin and demonstrates the effectiveness of SC injection in anatomically disparate tumor models. Mice bearing luciferase-expressing tumors underwent BLI after SC or IP injection of D-luciferin. The average time to maximal luminescence was 6 min (range 5–9 min) after SC injection and 8 min (range 5–8 min) after IP injection. Within 7 minutes of injection, SC and IP routes yielded similar luminescence in subcutaneous, intracranial, tongue, and lung xenograft tumor models. In a model of combined subcutaneous and intracranial xenografts, SC injection resulted in proportional luminescence at all sites, confirming that preferential delivery of substrate does not occur. While tumors were occasionally not visualized with IP injection, all tumors were visualized reliably with SC injection. Thus, SC injection of D-luciferin is a convenient and effective alternative to IP injection for BLI in nude mice. It may be a preferable approach, particularly for tumors with weaker signals and/or when greater precision is required.

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SALA-NEWBY, Graciela B., Catherine M. THOMSON, and Anthony K. CAMPBELL. "Sequence and biochemical similarities between the luciferases of the glow-worm Lampyris noctiluca and the firefly Photinus pyralis." Biochemical Journal 313, no. 3 (1996): 761–67. http://dx.doi.org/10.1042/bj3130761.

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A full-length clone encoding Lampyris noctiluca (British glow-worm) luciferase was isolated from a complementary DNA (cDNA) expression library constructed with mRNA extracted from light organs. The luciferase was a 547-residue protein, as deduced from the nucleotide sequence. The protein was closely related to those of other lampyrid beetles, the similarity to Photinus pyralis luciferase being 84% and to Luciola 67%. In contrast, Lampyris luciferase had less sequence similarity to the luciferases of the click beetle Pyrophorus, at 48%. Engineering Lampyris luciferase in vitro showed that the C-terminal peptide containing 12 amino acids in Photinus and 9 amino acids in Lampyris was essential for bioluminescence. The pH optimum and the Km values for ATP and luciferin were similar for both Photinus and Lampyris luciferases, although the light emitted by the latter shifted towards the blue and was less stable at 37 °C. It was concluded that the molecular and biochemical properties were not sufficient to explain the glowing or flashing of the two beetles Lampyris and Photinus.

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Barbaro, A., and S. Rożek. "Studies on luciferin-luciferase ATP assay in plants (etiolated wheat germs, and bean leaves)." Acta Societatis Botanicorum Poloniae 44, no. 3 (2015): 377–92. http://dx.doi.org/10.5586/asbp.1975.034.

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For ATP determination by the method of bioluminescence apparatus of home production was adapted from the equipment available in any isotope laboratory. The measurement error did not exceed 1.5 per cent. Methodical experiments concerned the choice of the extraction, fixation and storage methods of plant material for determination at the given moment of the amount of ATP in the tissues, unchanged by the analytical procedure. The highest ATP amounts were recovered by extraction with perchloric acid at high (25%) concentrations of the tissue in the homogenate. The best way of fixation of the material for later analyses was found to be freezing of ready extracts. Lyophilization and freezing of the plant material caused a several-fold decrease of the ATP level in the tissues. These results suggest the necessity of working in conditions of low temperature during the entire analytical procedure and strict observation of time limitation.

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Kieβling, M., K. Winsel, and B. Freytag. "Bioluminescence Reactions of Nucleotides Part 2: Investigations of AMP/ATP Solutions in the Luciferin/Luciferase System." Isotopenpraxis Isotopes in Environmental and Health Studies 27, no. 1 (1991): 42–44. http://dx.doi.org/10.1080/10256019108622462.

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Fukuba, Tatsuhiro, Takuroh Noguchi, Kei Okamura, and Teruo Fujii. "Adenosine Triphosphate Measurement in Deep Sea Using a Microfluidic Device." Micromachines 9, no. 8 (2018): 370. http://dx.doi.org/10.3390/mi9080370.

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Total ATP (adenosine triphosphate) concentration is a useful biochemical parameter for detecting microbial biomass or biogeochemical activity anomalies in the natural environment. In this study, we describe the development and evaluation of a new version of in situ ATP analyzer improved for the continuous and quantitative determination of ATP in submarine environments. We integrated a transparent microfluidic device containing a microchannel for cell lysis and a channel for the bioluminescence L–L (luciferin–luciferase) assay with a miniature pumping unit and a photometry module for the measurement of the bioluminescence intensity. A heater and a temperature sensor were also included in the system to maintain an optimal temperature for the L–L reaction. In this study, the analyzer was evaluated in deep sea environments, reaching a depth of 200 m using a remotely operated underwater vehicle. We show that the ATP analyzer successfully operated in the deep-sea environment and accurately quantified total ATP within the concentration lower than 5 × 10−11 M.

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Daydé, David, Florence Dommange, Stéphanie Lerondel, et al. "Quantitative Bioluminescent Method Appropriate for Bimodality Analysis To Improve and Follow a Syngenic Murine Model of B-Cell Lymphoma Expressing Human CD20." Blood 110, no. 11 (2007): 1400. http://dx.doi.org/10.1182/blood.v110.11.1400.1400.

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Abstract The anti-CD20 monoclonal antibody rituximab (C2B8) has shown promising results in the clinical treatment of patients with B-cell Non-Hodgkin’s Lymphoma (B-NHL). However, its therapeutic effect is variable whereas a better knowledge of factors affecting rituximab response could lead to improve its efficacy. It has been suggested that tumour burden could influence rituximab exposure and response in human. The lack of method to assess precisely tumour burden has however prevent to confirm such influence. Study of factors affecting antibody exposure requires bimodality analysis allowing to precise factors related to antibody from those related to tumour. We purposed to develop a quantitative method of tumour burden using bioluminescence imaging (BLI) and simultaneously scintigraphic study of monoclonal antibody biodistribution and exposure (indium-111 labelled ibritumomab, 2B8). Firstly, we developed a murine model of T-lymphoma cells transduced with human CD20 cDNA (EL4-hCD20) and transfected with luciferase plasmid (EL4-hCD20-luc). C57Bl6J mice and C57Bl6J TyrC2-J mice (albino phenotype) were injected with 8.10^3 EL4-hCD20-luc lymphoma cells by i.v. route. All mice died within 30 days with liver, spleen, bone marrow and lymph nodes involvement confirmed by immunohistochemistry and PCR analysis. In order to evaluate tumour growth and dissemination, potassium luciferin (2 or 4 mg) were injected bi-weekly intra peritoneally and tumour burden was analysed using Hamamatsu® CCD imaging system. Background noises and cosmic rays, which disturb the signal of bioluminescence, were eliminated from merging image before bioluminescent activity analysis. The sensitivity of analysis was below than 10^2 El4-hCD20-luc cells and there was proportionality between bioluminescent activity and cells number from 10^3 to 10^9 cells. The use of different binning allowed the follow-up of tumour burden evolution from 9 days after inoculation until the death of mice. We validated the used of black phenotype C57Bl6J compared to C57Bl6J TyrC2-J phenotype and demonstrated that the amount of luciferin could influence tumour growth. Regions of interest were delimited by segmentation method and we developed a reliable quantification method of bioluminescence activity taking into account absorption coefficients of the cells light emitted according to their localization. Such model and quantification method makes possible the follow-up of tumour growth and monoclonal antibody efficacy and exposure studied by bimodality analysis. D.D. is supported by a grant from Région Centre.

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Journal articles on the topic 'Luciferin-luciferase bioluminescence'

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