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Article

Chromophore of an Enhanced Green Fluorescent Protein Can Play a Photoprotective Role Due to Photobleaching

1
Division of Biophysics, Institute of Experimental Physics, Faculty of Physics, University of Warsaw, Pasteura 5, 02-093 Warsaw, Poland
2
Laboratory for Functional and Metabolic Imaging (LIFMET), Institute of Physics (IPHYS), School of Basic Sciences (SB), Ecole Polytechnique Fédérale de Lausanne (EPFL), CH-1015 Lausanne, Switzerland
3
Laboratory of Physics of Complex Matter (LPMC), Institute of Physics (IPHYS), School of Basic Sciences (SB), Ecole Polytechnique Fédérale de Lausanne (EPFL), CH-1015 Lausanne, Switzerland
4
Laboratory for Quantum Magnetism (LQM), Institute of Physics (IPHYS), School of Basic Sciences (SB), École Polytechnique Fédérale de Lausanne (EPFL), Station 3, CH-1015 Lausanne, Switzerland
5
ADSresonances, Route de Genève 60B, CH-1028 Préverenges, Switzerland
*
Authors to whom correspondence should be addressed.
Academic Editor: Eugene S. Vysotski
Int. J. Mol. Sci. 2021, 22(16), 8565; https://doi.org/10.3390/ijms22168565
Received: 18 June 2021 / Revised: 3 August 2021 / Accepted: 4 August 2021 / Published: 9 August 2021
Under stress conditions, elevated levels of cellular reactive oxygen species (ROS) may impair crucial cellular structures. To counteract the resulting oxidative damage, living cells are equipped with several defense mechanisms, including photoprotective functions of specific proteins. Here, we discuss the plausible ROS scavenging mechanisms by the enhanced green fluorescent protein, EGFP. To check if this protein could fulfill a photoprotective function, we employed electron spin resonance (ESR) in combination with spin-trapping. Two organic photosensitizers, rose bengal and methylene blue, as well as an inorganic photocatalyst, nano-TiO2, were used to photogenerate ROS. Spin-traps, TMP-OH and DMPO, and a nitroxide radical, TEMPOL, served as molecular targets for ROS. Our results show that EGFP quenches various forms of ROS, including superoxide radicals and singlet oxygen. Compared to the three proteins PNP, papain, and BSA, EGFP revealed high ROS quenching ability, which suggests its photoprotective role in living systems. Damage to the EGFP chromophore was also observed under strong photo-oxidative conditions. This study contributes to the discussion on the protective function of fluorescent proteins homologous to the green fluorescent protein (GFP). It also draws attention to the possible interactions of GFP-like proteins with ROS in systems where such proteins are used as biological markers. View Full-Text
Keywords: EGFP; photoprotection; superoxide radicals; singlet oxygen; scavenger; electron spin resonance; spin trapping; reactive oxygen species EGFP; photoprotection; superoxide radicals; singlet oxygen; scavenger; electron spin resonance; spin trapping; reactive oxygen species
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MDPI and ACS Style

Krasowska, J.; Pierzchała, K.; Bzowska, A.; Forró, L.; Sienkiewicz, A.; Wielgus-Kutrowska, B. Chromophore of an Enhanced Green Fluorescent Protein Can Play a Photoprotective Role Due to Photobleaching. Int. J. Mol. Sci. 2021, 22, 8565. https://doi.org/10.3390/ijms22168565

AMA Style

Krasowska J, Pierzchała K, Bzowska A, Forró L, Sienkiewicz A, Wielgus-Kutrowska B. Chromophore of an Enhanced Green Fluorescent Protein Can Play a Photoprotective Role Due to Photobleaching. International Journal of Molecular Sciences. 2021; 22(16):8565. https://doi.org/10.3390/ijms22168565

Chicago/Turabian Style

Krasowska, Joanna, Katarzyna Pierzchała, Agnieszka Bzowska, László Forró, Andrzej Sienkiewicz, and Beata Wielgus-Kutrowska. 2021. "Chromophore of an Enhanced Green Fluorescent Protein Can Play a Photoprotective Role Due to Photobleaching" International Journal of Molecular Sciences 22, no. 16: 8565. https://doi.org/10.3390/ijms22168565

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