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Analytical and Bioanalytical Chemistry

, Volume 410, Issue 26, pp 6837–6844 | Cite as

Protein-based fluorescent bioassay for low-dose gamma radiation exposures

  • Alena S. Petrova
  • Anna A. Lukonina
  • Dmitry V. Dementyev
  • Alexander Ya. Bolsunovsky
  • Anatoliy V. Popov
  • Nadezhda S. Kudryasheva
Research Paper

Abstract

The study suggests an application of a coelenteramide-containing fluorescent protein (CLM-CFP) as a simplest bioassay for gamma radiation exposures. “Discharged obelin,” a product of the bioluminescence reaction of the marine coelenterate Obelia longissima, was used as a representative of the CLM-CFP group. The bioassay is based on a simple enzymatic reaction—photochemical proton transfer in the coelenteramide-apoprotein complex. Components of this reaction differ in fluorescence color, providing, by this, an evaluation of the proton transfer efficiency in the photochemical process. This efficiency depends on the microenvironment of the coelenteramide within the protein complex, and, hence, can evaluate a destructive ability of gamma radiation. The CLM-CFP samples were exposed to gamma radiation (137Cs, 2 mGy/h) for 7 and 16 days at 20 °C and 5 °C, respectively. As a result, two fluorescence characteristics (overall fluorescence intensity and contributions of color components to the fluorescence spectra) were identified as bioassay parameters. Both parameters demonstrated high sensitivity of the CLM-CFP-based bioassay to the low-dose gamma radiation exposure (up to 100 mGy). Higher temperature (20 °C) enhanced the response of CLM-CFP to gamma radiation. This new bioassay can provide fluorescent multicolor assessment of protein destruction in cells and physiological liquids under exposure to low doses of gamma radiation.

Graphical abstract

Keywords

Bioassay Enzymes Fluorescence/luminescence Fluorescent protein Radiotoxicity Gamma radiation 

Notes

Acknowledgements

The authors would like to thank Alejandro D. Arroyo, University of Pennsylvania, for critical review of the manuscript.

Funding information

This work was supported by the state budget allocated to the fundamental research at the Russian Academy of Sciences, project 01201351504; the Russian Foundation for Basic Research, Grant No. 16-34-00695; and the Krasnoyarsk Regional Fund of Science and Technology Support. AVP’s research reported in this publication was supported by the National Center for Advancing Translational Sciences of the National Institutes of Health under award number UL1TR000003.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Alena S. Petrova
    • 1
  • Anna A. Lukonina
    • 2
  • Dmitry V. Dementyev
    • 3
  • Alexander Ya. Bolsunovsky
    • 3
  • Anatoliy V. Popov
    • 4
  • Nadezhda S. Kudryasheva
    • 2
    • 3
  1. 1.Krasnoyarsk State Agrarian UniversityKrasnoyarskRussia
  2. 2.Siberian Federal UniversityKrasnoyarskRussia
  3. 3.Institute of Biophysics SB RAS, FRC KSC SB RASKrasnoyarskRussia
  4. 4.Department of RadiologyUniversity of PennsylvaniaPhiladelphiaUSA

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