Protein-based fluorescent bioassay for low-dose gamma radiation exposures
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.
KeywordsBioassay Enzymes Fluorescence/luminescence Fluorescent protein Radiotoxicity Gamma radiation
The authors would like to thank Alejandro D. Arroyo, University of Pennsylvania, for critical review of the manuscript.
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.
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Conflict of interest
The authors declare that they have no conflict of interest.
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