Design and Biological Evaluation of hBest1-Containing Bilayer Nanostructures

Bestrophinopathies are a group of inherited retinal diseases caused by mutations in the <i>BEST1</i> gene. The protein encoded by this gene, bestorphin-1 (hBest1), is a calcium-dependent transmembrane channel localized on the basolateral membrane of retinal pigment epithelial (RPE) cells...

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Main Authors: Pavel Bakardzhiev, Teodora Koleva, Kirilka Mladenova, Pavel Videv, Veselina Moskova-Doumanova, Aleksander Forys, Sławomira Pusz, Tonya Andreeva, Svetla Petrova, Stanislav Rangelov, Jordan Doumanov
Format: Article
Language:English
Published: MDPI AG 2025-07-01
Series:Molecules
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Online Access:https://www.mdpi.com/1420-3049/30/14/2948
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author Pavel Bakardzhiev
Teodora Koleva
Kirilka Mladenova
Pavel Videv
Veselina Moskova-Doumanova
Aleksander Forys
Sławomira Pusz
Tonya Andreeva
Svetla Petrova
Stanislav Rangelov
Jordan Doumanov
author_facet Pavel Bakardzhiev
Teodora Koleva
Kirilka Mladenova
Pavel Videv
Veselina Moskova-Doumanova
Aleksander Forys
Sławomira Pusz
Tonya Andreeva
Svetla Petrova
Stanislav Rangelov
Jordan Doumanov
author_sort Pavel Bakardzhiev
collection DOAJ
description Bestrophinopathies are a group of inherited retinal diseases caused by mutations in the <i>BEST1</i> gene. The protein encoded by this gene, bestorphin-1 (hBest1), is a calcium-dependent transmembrane channel localized on the basolateral membrane of retinal pigment epithelial (RPE) cells. We have already demonstrated the surface behavior and organization of recombinant hBest1 and its interactions with membrane lipids such as 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), sphingomyelin (SM) and cholesterol (Chol) in models of biological membranes, which affect the hBest1 structure–function relationship. The main aim of our current investigation is to integrate pure hBest1 protein into lipid bilayer nanostructures. We synthesized and characterized various hBest1-containing nanostructures based on 1,2-Dipalmitoylphosphatidylcholine (DPPC), SM, glycerol monooleate (GMO) and Chol in different ratios and determined their cytotoxicity and incorporation into cell membranes and/or cells by immunofluorescence staining. Our results show that these newly designed nanoparticles are not cytotoxic and that their incorporation into MDCK II cell membranes (used as a model system) may provide a mechanism that could be applied to RPE cells expressing mutated hBest1 in order to restore their ion transport functions, affected by mutated and malfunctioning hBest1 molecules.
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publishDate 2025-07-01
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spelling doaj-art-15c7ae8f4bc54f26976de414f3a3958a2025-08-20T03:56:49ZengMDPI AGMolecules1420-30492025-07-013014294810.3390/molecules30142948Design and Biological Evaluation of hBest1-Containing Bilayer NanostructuresPavel Bakardzhiev0Teodora Koleva1Kirilka Mladenova2Pavel Videv3Veselina Moskova-Doumanova4Aleksander Forys5Sławomira Pusz6Tonya Andreeva7Svetla Petrova8Stanislav Rangelov9Jordan Doumanov10Institute of Polymers, Bulgarian Academy of Sciences, Akad. G. Bonchev Str. 103A, 1113 Sofia, BulgariaFaculty of Biology, Sofia University “St. Kliment Ohridski”, Dragan Tsankov Blvd., 1164 Sofia, BulgariaFaculty of Biology, Sofia University “St. Kliment Ohridski”, Dragan Tsankov Blvd., 1164 Sofia, BulgariaFaculty of Biology, Sofia University “St. Kliment Ohridski”, Dragan Tsankov Blvd., 1164 Sofia, BulgariaFaculty of Biology, Sofia University “St. Kliment Ohridski”, Dragan Tsankov Blvd., 1164 Sofia, BulgariaCentre of Polymer and Carbon Materials, Polish Academy of Sciences, ul. M. Curie-Skłodowskiej 34, 41-819 Zabrze, PolandCentre of Polymer and Carbon Materials, Polish Academy of Sciences, ul. M. Curie-Skłodowskiej 34, 41-819 Zabrze, PolandFaculty Life Sciences, Reutlingen University, Alteburgstraße 150, 72762 Reutlingen, GermanyFaculty of Biology, Sofia University “St. Kliment Ohridski”, Dragan Tsankov Blvd., 1164 Sofia, BulgariaInstitute of Polymers, Bulgarian Academy of Sciences, Akad. G. Bonchev Str. 103A, 1113 Sofia, BulgariaFaculty of Biology, Sofia University “St. Kliment Ohridski”, Dragan Tsankov Blvd., 1164 Sofia, BulgariaBestrophinopathies are a group of inherited retinal diseases caused by mutations in the <i>BEST1</i> gene. The protein encoded by this gene, bestorphin-1 (hBest1), is a calcium-dependent transmembrane channel localized on the basolateral membrane of retinal pigment epithelial (RPE) cells. We have already demonstrated the surface behavior and organization of recombinant hBest1 and its interactions with membrane lipids such as 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), sphingomyelin (SM) and cholesterol (Chol) in models of biological membranes, which affect the hBest1 structure–function relationship. The main aim of our current investigation is to integrate pure hBest1 protein into lipid bilayer nanostructures. We synthesized and characterized various hBest1-containing nanostructures based on 1,2-Dipalmitoylphosphatidylcholine (DPPC), SM, glycerol monooleate (GMO) and Chol in different ratios and determined their cytotoxicity and incorporation into cell membranes and/or cells by immunofluorescence staining. Our results show that these newly designed nanoparticles are not cytotoxic and that their incorporation into MDCK II cell membranes (used as a model system) may provide a mechanism that could be applied to RPE cells expressing mutated hBest1 in order to restore their ion transport functions, affected by mutated and malfunctioning hBest1 molecules.https://www.mdpi.com/1420-3049/30/14/2948hBest1bilayer nanostructuresMDCK II cells
spellingShingle Pavel Bakardzhiev
Teodora Koleva
Kirilka Mladenova
Pavel Videv
Veselina Moskova-Doumanova
Aleksander Forys
Sławomira Pusz
Tonya Andreeva
Svetla Petrova
Stanislav Rangelov
Jordan Doumanov
Design and Biological Evaluation of hBest1-Containing Bilayer Nanostructures
Molecules
hBest1
bilayer nanostructures
MDCK II cells
title Design and Biological Evaluation of hBest1-Containing Bilayer Nanostructures
title_full Design and Biological Evaluation of hBest1-Containing Bilayer Nanostructures
title_fullStr Design and Biological Evaluation of hBest1-Containing Bilayer Nanostructures
title_full_unstemmed Design and Biological Evaluation of hBest1-Containing Bilayer Nanostructures
title_short Design and Biological Evaluation of hBest1-Containing Bilayer Nanostructures
title_sort design and biological evaluation of hbest1 containing bilayer nanostructures
topic hBest1
bilayer nanostructures
MDCK II cells
url https://www.mdpi.com/1420-3049/30/14/2948
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