Modulation of Membrane Microviscosity by Protein-Mediated Carotenoid Delivery as Revealed by Time-Resolved Fluorescence Anisotropy
Carotenoids are potent antioxidants with a wide range of biomedical applications. However, their delivery into human cells is challenging and relatively inefficient. While the use of natural water-soluble carotenoproteins capable to reversibly bind carotenoids and transfer them into membranes is pro...
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| Format: | Article |
| Language: | English |
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MDPI AG
2022-09-01
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| Series: | Membranes |
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| Online Access: | https://www.mdpi.com/2077-0375/12/10/905 |
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| author | Alexey N. Semenov Danil A. Gvozdev Dmitry V. Zlenko Elena A. Protasova Anastasia R. Khashimova Evgenia Yu. Parshina Adil A. Baizhumanov Natalia Yu. Lotosh Eleonora E. Kim Yuriy N. Kononevich Alexey A. Pakhomov Alla A. Selishcheva Nikolai N. Sluchanko Evgeny A. Shirshin Eugene G. Maksimov |
| author_facet | Alexey N. Semenov Danil A. Gvozdev Dmitry V. Zlenko Elena A. Protasova Anastasia R. Khashimova Evgenia Yu. Parshina Adil A. Baizhumanov Natalia Yu. Lotosh Eleonora E. Kim Yuriy N. Kononevich Alexey A. Pakhomov Alla A. Selishcheva Nikolai N. Sluchanko Evgeny A. Shirshin Eugene G. Maksimov |
| author_sort | Alexey N. Semenov |
| collection | DOAJ |
| description | Carotenoids are potent antioxidants with a wide range of biomedical applications. However, their delivery into human cells is challenging and relatively inefficient. While the use of natural water-soluble carotenoproteins capable to reversibly bind carotenoids and transfer them into membranes is promising, the quantitative estimation of the delivery remains unclear. In the present work, we studied echinenone (ECN) delivery by cyanobacterial carotenoprotein AnaCTDH (C-terminal domain homolog of the Orange Carotenoid Protein from <i>Anabaena</i>), into liposome membranes labelled with BODIPY fluorescent probe. We observed that addition of AnaCTDH-ECN to liposomes led to the significant changes in the fast-kinetic component of the fluorescence decay curve, pointing on the dipole-dipole interactions between the probe and ECN within the membrane. It may serve as an indirect evidence of ECN delivery into membrane. To study the delivery in detail, we carried out molecular dynamics modeling of the localization of ECN within the lipid bilayer and calculate its orientation factor. Next, we exploited FRET to assess concentration of ECN delivered by AnaCTDH. Finally, we used time-resolved fluorescence anisotropy to assess changes in microviscosity of liposomal membranes. Incorporation of liposomes with β-carotene increased membrane microviscosity while the effect of astaxanthin and its mono- and diester forms was less pronounced. At temperatures below 30 °C addition of AnaCTDH-ECN increased membrane microviscosity in a concentration-dependent manner, supporting the protein-mediated carotenoid delivery mechanism. Combining all data, we propose FRET-based analysis and assessment of membrane microviscosity as potent approaches to characterize the efficiency of carotenoids delivery into membranes. |
| first_indexed | 2024-03-09T19:50:18Z |
| format | Article |
| id | doaj.art-93579752460a4033bda2f89f178b9264 |
| institution | Directory Open Access Journal |
| issn | 2077-0375 |
| language | English |
| last_indexed | 2024-03-09T19:50:18Z |
| publishDate | 2022-09-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Membranes |
| spelling | doaj.art-93579752460a4033bda2f89f178b92642023-11-24T01:12:27ZengMDPI AGMembranes2077-03752022-09-01121090510.3390/membranes12100905Modulation of Membrane Microviscosity by Protein-Mediated Carotenoid Delivery as Revealed by Time-Resolved Fluorescence AnisotropyAlexey N. Semenov0Danil A. Gvozdev1Dmitry V. Zlenko2Elena A. Protasova3Anastasia R. Khashimova4Evgenia Yu. Parshina5Adil A. Baizhumanov6Natalia Yu. Lotosh7Eleonora E. Kim8Yuriy N. Kononevich9Alexey A. Pakhomov10Alla A. Selishcheva11Nikolai N. Sluchanko12Evgeny A. Shirshin13Eugene G. Maksimov14Faculty of Biology, M.V. Lomonosov Moscow State University, 1-12 Leninskie Gory St., Moscow 119991, RussiaFaculty of Biology, M.V. Lomonosov Moscow State University, 1-12 Leninskie Gory St., Moscow 119991, RussiaFaculty of Biology, M.V. Lomonosov Moscow State University, 1-12 Leninskie Gory St., Moscow 119991, RussiaFaculty of Biology, M.V. Lomonosov Moscow State University, 1-12 Leninskie Gory St., Moscow 119991, RussiaFaculty of Biology, M.V. Lomonosov Moscow State University, 1-12 Leninskie Gory St., Moscow 119991, RussiaFaculty of Biology, M.V. Lomonosov Moscow State University, 1-12 Leninskie Gory St., Moscow 119991, RussiaFaculty of Biology, M.V. Lomonosov Moscow State University, 1-12 Leninskie Gory St., Moscow 119991, RussiaNational Research Center “Kurchatov Institute”, 1 Acad. Kurchatov Sq., Moscow 123182, RussiaA.N. Nesmeyanov Institute of Organoelement Compounds, Russian Academy of Sciences, Moscow 119991, RussiaA.N. Nesmeyanov Institute of Organoelement Compounds, Russian Academy of Sciences, Moscow 119991, RussiaA.N. Nesmeyanov Institute of Organoelement Compounds, Russian Academy of Sciences, Moscow 119991, RussiaFaculty of Biology, M.V. Lomonosov Moscow State University, 1-12 Leninskie Gory St., Moscow 119991, RussiaFederal Research Center of Biotechnology, Russian Academy of Sciences, 33 Leninsky Prospect, Moscow 119071, RussiaFaculty of Physics, M.V. Lomonosov Moscow State University, 1-2 Leninskie Gory St., Moscow 119991, RussiaFaculty of Biology, M.V. Lomonosov Moscow State University, 1-12 Leninskie Gory St., Moscow 119991, RussiaCarotenoids are potent antioxidants with a wide range of biomedical applications. However, their delivery into human cells is challenging and relatively inefficient. While the use of natural water-soluble carotenoproteins capable to reversibly bind carotenoids and transfer them into membranes is promising, the quantitative estimation of the delivery remains unclear. In the present work, we studied echinenone (ECN) delivery by cyanobacterial carotenoprotein AnaCTDH (C-terminal domain homolog of the Orange Carotenoid Protein from <i>Anabaena</i>), into liposome membranes labelled with BODIPY fluorescent probe. We observed that addition of AnaCTDH-ECN to liposomes led to the significant changes in the fast-kinetic component of the fluorescence decay curve, pointing on the dipole-dipole interactions between the probe and ECN within the membrane. It may serve as an indirect evidence of ECN delivery into membrane. To study the delivery in detail, we carried out molecular dynamics modeling of the localization of ECN within the lipid bilayer and calculate its orientation factor. Next, we exploited FRET to assess concentration of ECN delivered by AnaCTDH. Finally, we used time-resolved fluorescence anisotropy to assess changes in microviscosity of liposomal membranes. Incorporation of liposomes with β-carotene increased membrane microviscosity while the effect of astaxanthin and its mono- and diester forms was less pronounced. At temperatures below 30 °C addition of AnaCTDH-ECN increased membrane microviscosity in a concentration-dependent manner, supporting the protein-mediated carotenoid delivery mechanism. Combining all data, we propose FRET-based analysis and assessment of membrane microviscosity as potent approaches to characterize the efficiency of carotenoids delivery into membranes.https://www.mdpi.com/2077-0375/12/10/905carotenoproteinscarotenoid deliverymembrane microviscositytime-resolved fluorescence anisotropyFRETmolecular dynamics |
| spellingShingle | Alexey N. Semenov Danil A. Gvozdev Dmitry V. Zlenko Elena A. Protasova Anastasia R. Khashimova Evgenia Yu. Parshina Adil A. Baizhumanov Natalia Yu. Lotosh Eleonora E. Kim Yuriy N. Kononevich Alexey A. Pakhomov Alla A. Selishcheva Nikolai N. Sluchanko Evgeny A. Shirshin Eugene G. Maksimov Modulation of Membrane Microviscosity by Protein-Mediated Carotenoid Delivery as Revealed by Time-Resolved Fluorescence Anisotropy Membranes carotenoproteins carotenoid delivery membrane microviscosity time-resolved fluorescence anisotropy FRET molecular dynamics |
| title | Modulation of Membrane Microviscosity by Protein-Mediated Carotenoid Delivery as Revealed by Time-Resolved Fluorescence Anisotropy |
| title_full | Modulation of Membrane Microviscosity by Protein-Mediated Carotenoid Delivery as Revealed by Time-Resolved Fluorescence Anisotropy |
| title_fullStr | Modulation of Membrane Microviscosity by Protein-Mediated Carotenoid Delivery as Revealed by Time-Resolved Fluorescence Anisotropy |
| title_full_unstemmed | Modulation of Membrane Microviscosity by Protein-Mediated Carotenoid Delivery as Revealed by Time-Resolved Fluorescence Anisotropy |
| title_short | Modulation of Membrane Microviscosity by Protein-Mediated Carotenoid Delivery as Revealed by Time-Resolved Fluorescence Anisotropy |
| title_sort | modulation of membrane microviscosity by protein mediated carotenoid delivery as revealed by time resolved fluorescence anisotropy |
| topic | carotenoproteins carotenoid delivery membrane microviscosity time-resolved fluorescence anisotropy FRET molecular dynamics |
| url | https://www.mdpi.com/2077-0375/12/10/905 |
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