Toluidine Blue for the Determination of Binding of Anionic Polysaccharides to Lipid Raft Domains by Absorption Spectroscopy

The complexes of negatively charged polysaccharides with lipid vesicles have been shown to have applications in medicine, bioremediation, water purification, and construction of nano-biosensors. This article presents research on the formation of these complexes based on the interactions between thre...

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Main Authors: Sandra Gębczyńska, Julia Gdowska, Agata Mikos, Iga Gawrońska, Teresa Janas, Aleksander Czogalla, Tadeusz Janas
Format: Article
Language:English
Published: MDPI AG 2025-05-01
Series:Membranes
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Online Access:https://www.mdpi.com/2077-0375/15/5/139
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author Sandra Gębczyńska
Julia Gdowska
Agata Mikos
Iga Gawrońska
Teresa Janas
Aleksander Czogalla
Tadeusz Janas
author_facet Sandra Gębczyńska
Julia Gdowska
Agata Mikos
Iga Gawrońska
Teresa Janas
Aleksander Czogalla
Tadeusz Janas
author_sort Sandra Gębczyńska
collection DOAJ
description The complexes of negatively charged polysaccharides with lipid vesicles have been shown to have applications in medicine, bioremediation, water purification, and construction of nano-biosensors. This article presents research on the formation of these complexes based on the interactions between three types of liposomes, DOPC liposomes (which contain a lipid bilayer in the liquid-disordered (Ld) state), RAFT liposomes (which contain liquid-ordered (Lo) lipid raft domains surrounded by lipids in the Ld state) and SPH–CHL liposomes (which contain a lipid bilayer in the Lo state), and two selected anionic polysaccharides, polysialic acid (PSA) and polygalacturonic acid (PGA). The analysis was conducted using a toluidine blue (TB) probe and the absorption spectroscopy technique. In contrast to DOPC and SPH–CHL liposomes, binding of negatively charged PSA or PGA chains to RAFT liposomes induced a TB absorption maximum shift from 630 nm to 560 nm. The obtained results indicate that toluidine blue can be applied for monitoring the formation of these nano-complexes, and that the boundaries between Ld/Lo domains within membranes in RAFT liposomes can significantly enhance the binding affinity of negatively charged polysaccharides to the lipid bilayer surface. The observed metachromatic shift in TB absorption suggests that negatively charged PSA and PGA chains interact with the Ld/Lo boundaries within RAFT liposome membranes.
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spelling doaj-art-8fa24fbea0344ed8bebc33276d068aad2025-08-20T01:56:17ZengMDPI AGMembranes2077-03752025-05-0115513910.3390/membranes15050139Toluidine Blue for the Determination of Binding of Anionic Polysaccharides to Lipid Raft Domains by Absorption SpectroscopySandra Gębczyńska0Julia Gdowska1Agata Mikos2Iga Gawrońska3Teresa Janas4Aleksander Czogalla5Tadeusz Janas6Institute of Biology, University of Opole, Kominka 6, 45-032 Opole, PolandInstitute of Biology, University of Opole, Kominka 6, 45-032 Opole, PolandInstitute of Biology, University of Opole, Kominka 6, 45-032 Opole, PolandInstitute of Biology, University of Opole, Kominka 6, 45-032 Opole, PolandInstitute of Biology, University of Opole, Kominka 6, 45-032 Opole, PolandDepartment of Cytobiochemistry, Faculty of Biotechnology, University of Wroclaw, F. Joliot-Curie 14a, 50-383 Wrocław, PolandInstitute of Biology, University of Opole, Kominka 6, 45-032 Opole, PolandThe complexes of negatively charged polysaccharides with lipid vesicles have been shown to have applications in medicine, bioremediation, water purification, and construction of nano-biosensors. This article presents research on the formation of these complexes based on the interactions between three types of liposomes, DOPC liposomes (which contain a lipid bilayer in the liquid-disordered (Ld) state), RAFT liposomes (which contain liquid-ordered (Lo) lipid raft domains surrounded by lipids in the Ld state) and SPH–CHL liposomes (which contain a lipid bilayer in the Lo state), and two selected anionic polysaccharides, polysialic acid (PSA) and polygalacturonic acid (PGA). The analysis was conducted using a toluidine blue (TB) probe and the absorption spectroscopy technique. In contrast to DOPC and SPH–CHL liposomes, binding of negatively charged PSA or PGA chains to RAFT liposomes induced a TB absorption maximum shift from 630 nm to 560 nm. The obtained results indicate that toluidine blue can be applied for monitoring the formation of these nano-complexes, and that the boundaries between Ld/Lo domains within membranes in RAFT liposomes can significantly enhance the binding affinity of negatively charged polysaccharides to the lipid bilayer surface. The observed metachromatic shift in TB absorption suggests that negatively charged PSA and PGA chains interact with the Ld/Lo boundaries within RAFT liposome membranes.https://www.mdpi.com/2077-0375/15/5/139absorption spectroscopyliposomespolysialic acidpolygalacturonic acidlipid raftstoluidine blue
spellingShingle Sandra Gębczyńska
Julia Gdowska
Agata Mikos
Iga Gawrońska
Teresa Janas
Aleksander Czogalla
Tadeusz Janas
Toluidine Blue for the Determination of Binding of Anionic Polysaccharides to Lipid Raft Domains by Absorption Spectroscopy
Membranes
absorption spectroscopy
liposomes
polysialic acid
polygalacturonic acid
lipid rafts
toluidine blue
title Toluidine Blue for the Determination of Binding of Anionic Polysaccharides to Lipid Raft Domains by Absorption Spectroscopy
title_full Toluidine Blue for the Determination of Binding of Anionic Polysaccharides to Lipid Raft Domains by Absorption Spectroscopy
title_fullStr Toluidine Blue for the Determination of Binding of Anionic Polysaccharides to Lipid Raft Domains by Absorption Spectroscopy
title_full_unstemmed Toluidine Blue for the Determination of Binding of Anionic Polysaccharides to Lipid Raft Domains by Absorption Spectroscopy
title_short Toluidine Blue for the Determination of Binding of Anionic Polysaccharides to Lipid Raft Domains by Absorption Spectroscopy
title_sort toluidine blue for the determination of binding of anionic polysaccharides to lipid raft domains by absorption spectroscopy
topic absorption spectroscopy
liposomes
polysialic acid
polygalacturonic acid
lipid rafts
toluidine blue
url https://www.mdpi.com/2077-0375/15/5/139
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