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Fluorinated synthetic anion carriers: experimental and computational insights into transmembrane chloride transport

Fluorinated synthetic anion carriers: experimental and computational insights into transmembrane chloride transport
Fluorinated synthetic anion carriers: experimental and computational insights into transmembrane chloride transport
A series of fluorinated tripodal tris-thioureas function as highly active anion transporters across lipid bilayers and cell membranes. Here, we investigate their mechanism of action using anion transport assays in cells and synthetic vesicles and molecular modelling of transporter–lipid interactions. When compared with non-fluorinated analogues, fluorinated compounds demonstrate a different mechanism of membrane transport because the free transporter cannot effectively diffuse through the membrane. As a result, in H+/Cl− cotransport assays, fluorinated transporters require the presence of oleic acid to form anionic oleate complexes for recycling of the transporter, whereas non-fluorinated analogues readily diffuse through the membrane as free transporters and show synergistic transport with the proton transporter gramicidin. Molecular dynamics simulations revealed markedly stronger transporter–lipid interactions for fluorinated compounds compared with non-fluorinated analogues and hence, higher energy barriers for fluorinated compounds to cross the membrane as free transporters. With use of appropriate proton transporters to ensure measurement of the correct rate-limiting steps, the transport rates determined in synthetic vesicle assays show excellent agreement with the anion transport rates determined in cell-based assays. We conclude that integration of computational and experimental methods provides a strategy to optimise transmembrane anion transporter design for biomedical applications.
1478-6524
Spooner, Michael
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Li, Hongyu
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Marques, Igor
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Costa, Pedro M.R.
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Wu, Xin
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Howe, Ethan NW
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Busschaert, Nathalie
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Moore, Stephen
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Light, Mark
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Sheppard, David N.
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Félix, Vítor
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Gale, Philip
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Spooner, Michael
b2fc20ec-22e1-42f9-82b8-5afd2666c25d
Li, Hongyu
ecf34b99-1d3d-49c1-be7b-5edee54a6a2e
Marques, Igor
97cbc26e-6d94-43e6-a934-1d69dad8de06
Costa, Pedro M.R.
a956ddb9-002e-4c69-aa85-c0dafe913cdb
Wu, Xin
7c9d888f-abe1-4332-a26a-c850d220164c
Howe, Ethan NW
be05ae9a-02e0-443b-97b2-d65cc387af40
Busschaert, Nathalie
9abe79c0-7540-4677-bf27-84e8e0e048f6
Moore, Stephen
e7a4f98a-ac24-4d29-b1b7-f9ef32d7a63e
Light, Mark
cf57314e-6856-491b-a8d2-2dffc452e161
Sheppard, David N.
d68744b7-e8e1-4388-9a92-39f49f9c817e
Félix, Vítor
b4098ac2-b86b-48f3-af11-c767e24fac6e
Gale, Philip
c840b7e9-6847-4843-91af-fa0f8563d943

Spooner, Michael, Li, Hongyu, Marques, Igor, Costa, Pedro M.R., Wu, Xin, Howe, Ethan NW, Busschaert, Nathalie, Moore, Stephen, Light, Mark, Sheppard, David N., Félix, Vítor and Gale, Philip (2018) Fluorinated synthetic anion carriers: experimental and computational insights into transmembrane chloride transport. Chemical Science. (doi:10.1039/C8SC05155K).

Record type: Article

Abstract

A series of fluorinated tripodal tris-thioureas function as highly active anion transporters across lipid bilayers and cell membranes. Here, we investigate their mechanism of action using anion transport assays in cells and synthetic vesicles and molecular modelling of transporter–lipid interactions. When compared with non-fluorinated analogues, fluorinated compounds demonstrate a different mechanism of membrane transport because the free transporter cannot effectively diffuse through the membrane. As a result, in H+/Cl− cotransport assays, fluorinated transporters require the presence of oleic acid to form anionic oleate complexes for recycling of the transporter, whereas non-fluorinated analogues readily diffuse through the membrane as free transporters and show synergistic transport with the proton transporter gramicidin. Molecular dynamics simulations revealed markedly stronger transporter–lipid interactions for fluorinated compounds compared with non-fluorinated analogues and hence, higher energy barriers for fluorinated compounds to cross the membrane as free transporters. With use of appropriate proton transporters to ensure measurement of the correct rate-limiting steps, the transport rates determined in synthetic vesicle assays show excellent agreement with the anion transport rates determined in cell-based assays. We conclude that integration of computational and experimental methods provides a strategy to optimise transmembrane anion transporter design for biomedical applications.

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Accepted/In Press date: 5 December 2018
e-pub ahead of print date: 14 December 2018

Identifiers

Local EPrints ID: 427210
URI: http://eprints.soton.ac.uk/id/eprint/427210
ISSN: 1478-6524
PURE UUID: b208e1e2-2cab-48d7-9097-f4f2c53e7ce3
ORCID for Mark Light: ORCID iD orcid.org/0000-0002-0585-0843
ORCID for Philip Gale: ORCID iD orcid.org/0000-0001-9751-4910

Catalogue record

Date deposited: 08 Jan 2019 17:30
Last modified: 26 Nov 2021 02:43

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Contributors

Author: Michael Spooner
Author: Hongyu Li
Author: Igor Marques
Author: Pedro M.R. Costa
Author: Xin Wu
Author: Ethan NW Howe
Author: Nathalie Busschaert
Author: Stephen Moore
Author: Mark Light ORCID iD
Author: David N. Sheppard
Author: Vítor Félix
Author: Philip Gale ORCID iD

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