Chromophore of an Enhanced Green Fluorescent Protein Can Play a Photoprotective Role Due to Photobleaching
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 discu...
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MDPI AG
2021-08-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/22/16/8565 |
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| author | Joanna Krasowska Katarzyna Pierzchała Agnieszka Bzowska László Forró Andrzej Sienkiewicz Beata Wielgus-Kutrowska |
| author_facet | Joanna Krasowska Katarzyna Pierzchała Agnieszka Bzowska László Forró Andrzej Sienkiewicz Beata Wielgus-Kutrowska |
| author_sort | Joanna Krasowska |
| collection | DOAJ |
| description | 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-TiO<sub>2</sub>, 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. |
| first_indexed | 2024-03-10T08:45:09Z |
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| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T08:45:09Z |
| publishDate | 2021-08-01 |
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| series | International Journal of Molecular Sciences |
| spelling | doaj.art-ca066cf86f12492cae2a0f14878d44e12023-11-22T07:57:02ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672021-08-012216856510.3390/ijms22168565Chromophore of an Enhanced Green Fluorescent Protein Can Play a Photoprotective Role Due to PhotobleachingJoanna Krasowska0Katarzyna Pierzchała1Agnieszka Bzowska2László Forró3Andrzej Sienkiewicz4Beata Wielgus-Kutrowska5Division of Biophysics, Institute of Experimental Physics, Faculty of Physics, University of Warsaw, Pasteura 5, 02-093 Warsaw, PolandLaboratory for Functional and Metabolic Imaging (LIFMET), Institute of Physics (IPHYS), School of Basic Sciences (SB), Ecole Polytechnique Fédérale de Lausanne (EPFL), CH-1015 Lausanne, SwitzerlandDivision of Biophysics, Institute of Experimental Physics, Faculty of Physics, University of Warsaw, Pasteura 5, 02-093 Warsaw, PolandLaboratory of Physics of Complex Matter (LPMC), Institute of Physics (IPHYS), School of Basic Sciences (SB), Ecole Polytechnique Fédérale de Lausanne (EPFL), CH-1015 Lausanne, SwitzerlandLaboratory of Physics of Complex Matter (LPMC), Institute of Physics (IPHYS), School of Basic Sciences (SB), Ecole Polytechnique Fédérale de Lausanne (EPFL), CH-1015 Lausanne, SwitzerlandDivision of Biophysics, Institute of Experimental Physics, Faculty of Physics, University of Warsaw, Pasteura 5, 02-093 Warsaw, PolandUnder 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-TiO<sub>2</sub>, 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.https://www.mdpi.com/1422-0067/22/16/8565EGFPphotoprotectionsuperoxide radicalssinglet oxygenscavengerelectron spin resonance |
| spellingShingle | Joanna Krasowska Katarzyna Pierzchała Agnieszka Bzowska László Forró Andrzej Sienkiewicz Beata Wielgus-Kutrowska Chromophore of an Enhanced Green Fluorescent Protein Can Play a Photoprotective Role Due to Photobleaching International Journal of Molecular Sciences EGFP photoprotection superoxide radicals singlet oxygen scavenger electron spin resonance |
| title | Chromophore of an Enhanced Green Fluorescent Protein Can Play a Photoprotective Role Due to Photobleaching |
| title_full | Chromophore of an Enhanced Green Fluorescent Protein Can Play a Photoprotective Role Due to Photobleaching |
| title_fullStr | Chromophore of an Enhanced Green Fluorescent Protein Can Play a Photoprotective Role Due to Photobleaching |
| title_full_unstemmed | Chromophore of an Enhanced Green Fluorescent Protein Can Play a Photoprotective Role Due to Photobleaching |
| title_short | Chromophore of an Enhanced Green Fluorescent Protein Can Play a Photoprotective Role Due to Photobleaching |
| title_sort | chromophore of an enhanced green fluorescent protein can play a photoprotective role due to photobleaching |
| topic | EGFP photoprotection superoxide radicals singlet oxygen scavenger electron spin resonance |
| url | https://www.mdpi.com/1422-0067/22/16/8565 |
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