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Bioluminescent test systems based on firefly luciferase for studying stress effects on living cells

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Abstract

The bioluminescent luciferin-luciferase reaction is based on the oxidation of D-luciferin by oxygen in the presence of ATP and magnesium ions, catalyzed by firefly luciferase. The possibilities of using this reaction to study the influence of external effectors of a physical and chemical nature (temperature exposure, additions of drugs, membrane-active compounds, etc.) on living cells (prokaryotes and eukaryotes) are considered. Examples of the use of test systems based on living cells producing thermostable firefly luciferase for monitoring cellular homeostasis are given. The study of the kinetics of changes in the concentration of ATP and luciferase inside and outside cells made it possible to determine in dynamics the metabolic activity, cytotoxicity, and survival of cells under conditions of cellular stress, to study the processes of ATP synthesis/hydrolysis, and to evaluate the effectiveness of lytic agents in changing the permeability of the cell membrane.

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References

  • Bajerski F, Stock J, Hanf B, Darienko T, Heine-Dobbernack E, Lorenz M, Naujox L, Keller ERJ, Schumacher HM, Friedl T, Eberth S, Mock H-P, Kniemeyer O, Overmann J (2018) ATP content and cell viability as indicators for cryostress across the diversity of life. Front Physiol 9:921. https://doi.org/10.3389/fphys.2018.00921

  • Braissant O, Astasov-Frauenhoffer M, Waltimo T, Bonkat G (2020) A review of methods to determine viability, vitality, and metabolic rates in microbiology. Front Microbiol 11:547458. https://doi.org/10.3389/fmicb.2020.547458

  • Fernández-Piñas F, Rodea-Palomares I, Leganés F, González-Pleiter M, Muñoz-Martín MA (2014) Evaluation of the ecotoxicity of pollutants with bioluminescent microorganisms. In: Thouand G, Marks R (eds) Bioluminescence: fundamentals and applications in biotechnology, vol 2. Springer, Berlin, Heidelberg, pp 65–135. https://doi.org/10.1007/978-3-662-43619-6_3

  • Ihssen J, Jovanovic N, Sirec T, Spitz U (2021) Real-time monitoring of extracellular ATP in bacterial cultures using thermostable luciferase. PLoS One 16:e0244200. https://doi.org/10.1371/journal.pone.0244200

  • Koksharov MI, Ugarova NN (2011) Thermostabilization of firefly luciferase by in vivo directed evolution. Prot Engin Des Selec 24:835–844. https://doi.org/10.1093/protein/gzr044

  • Kudryasheva NS (2006) Bioluminescence and exogenous compounds: physico-chemical basis for bioluminescent assay. J Photochem Photobiol B Biol 83:77–86. https://doi.org/10.1016/j.jphotobiol.2005.10.003

  • Kudryasheva NS, Kovel ES (2019) Monitoring of low-intensity exposures via luminescent bioassays of different complexity: cells, enzyme reactions, and fluorescent proteins. Int J Mol Sci 20:4451. https://doi.org/10.3390/ijms20184451

  • Kudryasheva NS, Rozhko TV (2015) Effect of low-dose ionizing radiation on luminous marine bacteria: radiation hormesis and toxicity. J Environ Radioact 142:68e77. https://doi.org/10.1016/j.jenvrad.2015.01.012

  • Leuenberger P, Ganscha S, Kahraman A, Cappelletti V, Boersema PJ, Mering C, Claassen M, Picotti P (2017) Cell-wide analysis of protein thermal unfolding reveals determinants of thermostability. Science 355:eaai7825. https://doi.org/10.1126/science.aai7825

  • Lomakina GY, Ugarova NN (2022a) Kinetics of the interaction of colistin with live Escherichia coli cells by the bioluminescence method. Moscow Univer Chem Bull 77(1):42–47. https://doi.org/10.3103/S0027131422010059

    Article  Google Scholar 

  • Lomakina GY, Ugarova NN (2022b) Application of bioluminescent methods to study the effect of the membrane-active antibiotic colistin on bacterial cells. Photochem Photobiol. https://doi.org/10.1111/php.13606

    Article  Google Scholar 

  • Lomakina GY, Modestova Y, Ugarova NN (2015) Bioluminescence assay for cell viability. Biochemistry (Moscow) 80(6):701–713. https://doi.org/10.1134/s0006297915060061

  • Lomakina GY, Koryagina VA, Ugarova NN (2018) Firefly luciferase as an intracellular marker of bacterial temperature stress (in russ). Russ J BiolPhys Chem 3(2):441–446

    Google Scholar 

  • Lomakina GY, Konik PA, Ugarova NN (2020a) The kinetics of hydrolysis of ATP by apyrase A from solanum tuberosum. Moscow Univer Chem Bull 75(6):1–8. https://doi.org/10.3103/S0027131420060139

  • Lomakina GY, Fomina AD, Ugarova NN (2020b) Kinetics of interaction of digitonin and its analogues with HEK293 cells studied by the bioluminescence method. Moscow Univer Chem Bull 75(3):186–194. https://doi.org/10.3103/S0027131420030086

  • Mempin R, Tran H, Chen C, Gong H, Ho KK, Lu S (2013) Release of extracellular ATP by bacteria during growth. BMC Microbiol 13:301. https://doi.org/10.1186/1471-2180-13-301

  • Moubareck CA (2020) Polymyxins and bacterial membranes: a review of antibacterial activity and mechanisms of resistance. Membranes 10:181. https://doi.org/10.3390/membranes10080181

  • Sabnis A, Hagart KL, Klöckner A, Becce M, Evans LE, Furniss RCD, Mavridou DA, Murphy R, Stevens MM, Davies JC, Larrouy-Maumus GJ, Clarke TB, Edwards AM (2021) Colistin kills bacteria by targeting lipopolysaccharide in the cytoplasmic membrane. Elife 10:e65836. https://doi.org/10.7554/eLife.65836

  • Shan Y, Gandt AB, Row SE, Deisinger JP, Conlon BP, Lewis K (2017) ATP-dependent persister formation in Escherichia coli. Mbio 8:e02267–e02316. https://doi.org/10.1128/mbio.02267-16

  • Smelt JPPM, Brul S (2014) Thermal inactivation of microorganisms. Crit Rev Food Sci Nutr 54:1371–1385. https://doi.org/10.1080/10408398.2011.637645

  • Syed AJ, Anderson JC (2021) Applications of bioluminescence in biotechnology and beyond. Chem Soc Rev 50:5668–5705. https://doi.org/10.1039/D0CS01492C

  • Ugarova NN, Lomakina GY (2017) Bioluminescence methods for the rapid assay of the specific activity of lyophilized BCG vaccine. Adv Tech Biol Med 5:243

    Article  Google Scholar 

  • Ugarova NN, Koksharov MI, Lomakina GY (2009) Reagent for determination of adenosine-5`-triphosphate. RF Patent 2420594

  • Ugarova NN, Lomakina GY, Modestova Y, Chernikov SV, Vinokurova NV, Otrashevskaya EV, Gorbachev VY (2016) A simplified ATP method for the rapid control of cell viability in a freeze-dried BCG vaccine. J Microbiol Meth 130:48–53. https://doi.org/10.1016/j.mimet.2016.08.027

  • Ugarova NN, Lomakina GY, Perevyshina TA, Otrashevskaya EV, Chernikov SV (2019) Controlling BCG vaccine’s cell viability in the process of its production by an bioluminescent ATP assay. Moscow Univer Chem Bull 74(4):191–197. https://doi.org/10.3103/S0027131419040084

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Funding

This work was performed as part of the state registration topic of Lomonosov Moscow State University, no. 121041500039–8

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Correspondence to Galina Yu. Lomakina.

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Lomakina, G.Y., Ugarova, N.N. Bioluminescent test systems based on firefly luciferase for studying stress effects on living cells. Biophys Rev 14, 887–892 (2022). https://doi.org/10.1007/s12551-022-00978-y

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