The Impact of an Anchoring Layer on the Formation of Tethered Bilayer Lipid Membranes on Silver Substrates

Tethered bilayer lipid membranes (tBLMs) have been known as stable and versatile experimental platforms for protein–membrane interaction studies. In this work, the assembly of functional tBLMs on silver substrates and the effect of the molecular chain-length of backfiller molecules on their properti...

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Autores principales:	Indrė Aleknavičienė, Martynas Talaikis, Rima Budvytyte, Gintaras Valincius
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	tethered bilayer lipid membranes self-assembled monolayers silver films molecular anchors electrochemical impedance spectroscopy surface-enhanced Raman scattering Organic chemistry QD241-441
Acceso en línea:	https://doaj.org/article/63398dd4fb5f47e6af457cafc2892426
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spelling	oai:doaj.org-article:63398dd4fb5f47e6af457cafc28924262021-11-25T18:28:02ZThe Impact of an Anchoring Layer on the Formation of Tethered Bilayer Lipid Membranes on Silver Substrates10.3390/molecules262268781420-3049https://doaj.org/article/63398dd4fb5f47e6af457cafc28924262021-11-01T00:00:00Zhttps://www.mdpi.com/1420-3049/26/22/6878https://doaj.org/toc/1420-3049Tethered bilayer lipid membranes (tBLMs) have been known as stable and versatile experimental platforms for protein–membrane interaction studies. In this work, the assembly of functional tBLMs on silver substrates and the effect of the molecular chain-length of backfiller molecules on their properties were investigated. The following backfillers 3-mercapto-1-propanol (3M1P), 4-mercapto-1-butanol (4M1B), 6-mercapto-1-hexanol (6M1H), and 9-mercapto-1-nonanol (9M1N) mixed with the molecular anchor WC14 (20-tetradecyloxy-3,6,9,12,15,18,22 heptaoxahexatricontane-1-thiol) were used to form self-assembled monolayers (SAMs) on silver, which influenced a fusion of multilamellar vesicles and the formation of tBLMs. Spectroscopic analysis by SERS and RAIRS has shown that by using different-length backfiller molecules, it is possible to control WC14 anchor molecules orientation on the surface. An introduction of increasingly longer surface backfillers in the mixed SAM may be related to the increasing SAMs molecular order and more vertical orientation of WC14 at both the hydrophilic ethylenoxide segment and the hydrophobic lipid bilayer anchoring alkane chains. Since no clustering of WC14 alkane chains, which is deleterious for tBLM integrity, was observed on dry samples, the suitability of mixed-component SAMs for subsequent tBLM formation was further interrogated by electrochemical impedance spectroscopy (EIS). EIS showed the arrangement of well-insulating tBLMs if 3M1P was used as a backfiller. An increase in the length of the backfiller led to increased defectiveness of tBLMs. Despite variable defectiveness, all tBLMs responded to the pore-forming cholesterol-dependent cytolysin, vaginolysin in a manner consistent with the functional reconstitution of the toxin into phospholipid bilayer. This experiment demonstrates the biological relevance of tBLMs assembled on silver surfaces and indicates their utility as biosensing elements for the detection of pore-forming toxins in liquid samples.Indrė AleknavičienėMartynas TalaikisRima BudvytyteGintaras ValinciusMDPI AGarticletethered bilayer lipid membranesself-assembled monolayerssilver filmsmolecular anchorselectrochemical impedance spectroscopysurface-enhanced Raman scatteringOrganic chemistryQD241-441ENMolecules, Vol 26, Iss 6878, p 6878 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	tethered bilayer lipid membranes self-assembled monolayers silver films molecular anchors electrochemical impedance spectroscopy surface-enhanced Raman scattering Organic chemistry QD241-441
spellingShingle	tethered bilayer lipid membranes self-assembled monolayers silver films molecular anchors electrochemical impedance spectroscopy surface-enhanced Raman scattering Organic chemistry QD241-441 Indrė Aleknavičienė Martynas Talaikis Rima Budvytyte Gintaras Valincius The Impact of an Anchoring Layer on the Formation of Tethered Bilayer Lipid Membranes on Silver Substrates
description	Tethered bilayer lipid membranes (tBLMs) have been known as stable and versatile experimental platforms for protein–membrane interaction studies. In this work, the assembly of functional tBLMs on silver substrates and the effect of the molecular chain-length of backfiller molecules on their properties were investigated. The following backfillers 3-mercapto-1-propanol (3M1P), 4-mercapto-1-butanol (4M1B), 6-mercapto-1-hexanol (6M1H), and 9-mercapto-1-nonanol (9M1N) mixed with the molecular anchor WC14 (20-tetradecyloxy-3,6,9,12,15,18,22 heptaoxahexatricontane-1-thiol) were used to form self-assembled monolayers (SAMs) on silver, which influenced a fusion of multilamellar vesicles and the formation of tBLMs. Spectroscopic analysis by SERS and RAIRS has shown that by using different-length backfiller molecules, it is possible to control WC14 anchor molecules orientation on the surface. An introduction of increasingly longer surface backfillers in the mixed SAM may be related to the increasing SAMs molecular order and more vertical orientation of WC14 at both the hydrophilic ethylenoxide segment and the hydrophobic lipid bilayer anchoring alkane chains. Since no clustering of WC14 alkane chains, which is deleterious for tBLM integrity, was observed on dry samples, the suitability of mixed-component SAMs for subsequent tBLM formation was further interrogated by electrochemical impedance spectroscopy (EIS). EIS showed the arrangement of well-insulating tBLMs if 3M1P was used as a backfiller. An increase in the length of the backfiller led to increased defectiveness of tBLMs. Despite variable defectiveness, all tBLMs responded to the pore-forming cholesterol-dependent cytolysin, vaginolysin in a manner consistent with the functional reconstitution of the toxin into phospholipid bilayer. This experiment demonstrates the biological relevance of tBLMs assembled on silver surfaces and indicates their utility as biosensing elements for the detection of pore-forming toxins in liquid samples.
format	article
author	Indrė Aleknavičienė Martynas Talaikis Rima Budvytyte Gintaras Valincius
author_facet	Indrė Aleknavičienė Martynas Talaikis Rima Budvytyte Gintaras Valincius
author_sort	Indrė Aleknavičienė
title	The Impact of an Anchoring Layer on the Formation of Tethered Bilayer Lipid Membranes on Silver Substrates
title_short	The Impact of an Anchoring Layer on the Formation of Tethered Bilayer Lipid Membranes on Silver Substrates
title_full	The Impact of an Anchoring Layer on the Formation of Tethered Bilayer Lipid Membranes on Silver Substrates
title_fullStr	The Impact of an Anchoring Layer on the Formation of Tethered Bilayer Lipid Membranes on Silver Substrates
title_full_unstemmed	The Impact of an Anchoring Layer on the Formation of Tethered Bilayer Lipid Membranes on Silver Substrates
title_sort	impact of an anchoring layer on the formation of tethered bilayer lipid membranes on silver substrates
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/63398dd4fb5f47e6af457cafc2892426
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The Impact of an Anchoring Layer on the Formation of Tethered Bilayer Lipid Membranes on Silver Substrates

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