An increased throughput workflow to identify ion transport and membrane lysis agents for antimicrobial discovery
An increased throughput workflow to identify ion transport and membrane lysis agents for antimicrobial discovery
Small molecule ion transporters have shown promise as potential therapeutics for microbial infections, cancer and channelopathies. However, there are still gaps in our understanding of how ion transport function in model vesicle membranes translates to cell membranes of interest. The lipid composition of the membranes of bacterial and cancer cells differs markedly from normal human cells, yet the influence of the lipid composition on membrane function is rarely investigated – in part because of the low throughput and high cost of ion transport experiments. Here, we report an increased throughput Workflow to identify biologically relevant, pH-driven ion transport and membrane lysis pathways in vesicle membranes. We developed a set of four assays designed to report on different transport and lysis processes. We validated our assays against a panel of known transporters and produced a stepwise Workflow for the evaluation of libraries of compounds. We applied our Workflow to screen a library (Library 1) of 31 supramolecular, self-associating amphiphiles (SSAs) for transport and lysis activity in a range of vesicles with different lipid compositions, designed to mimic different types of cells, and consequently identified seven promising transporters. Antimicrobial experiments found that six of these promising transporters showed good antimicrobial activity against clinically relevant Staphylococcus aureus strains, highlighting the promise of our Workflow in identifying potential antimicrobial agents. We then applied our Workflow to investigate the ion transport and lysis properties of a second library (Library 2) of SSAs with established antimicrobial and anticancer properties, aiming to provide insight into the biological modes of action.
Yang, Kylie
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Marsh, Caleb
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White, Lisa J.
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Molyneux, Fergus W.
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Allam, Thomas L.
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Popoola, Precious I.A.
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Keers, Olivia B.
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Rice, Matthew
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Hilton, Kira L.F.
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Kotak, Hiral A.
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Sutton, J. Mark
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Ortega-Roldan, Jose L.
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Hind, Charlotte K.
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Hiscock, Jennifer R.
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Haynes, Cally J.E.
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Yang, Kylie
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Marsh, Caleb
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White, Lisa J.
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Molyneux, Fergus W.
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Allam, Thomas L.
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Popoola, Precious I.A.
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Keers, Olivia B.
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Rice, Matthew
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Hilton, Kira L.F.
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Kotak, Hiral A.
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Sutton, J. Mark
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Ortega-Roldan, Jose L.
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Hind, Charlotte K.
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Hiscock, Jennifer R.
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Haynes, Cally J.E.
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Yang, Kylie, Marsh, Caleb, White, Lisa J., Molyneux, Fergus W., Allam, Thomas L., Popoola, Precious I.A., Keers, Olivia B., Rice, Matthew, Hilton, Kira L.F., Kotak, Hiral A., Sutton, J. Mark, Ortega-Roldan, Jose L., Hind, Charlotte K., Hiscock, Jennifer R. and Haynes, Cally J.E.
(2026)
An increased throughput workflow to identify ion transport and membrane lysis agents for antimicrobial discovery.
Chemical Science.
(doi:10.1039/d5sc09781a).
Abstract
Small molecule ion transporters have shown promise as potential therapeutics for microbial infections, cancer and channelopathies. However, there are still gaps in our understanding of how ion transport function in model vesicle membranes translates to cell membranes of interest. The lipid composition of the membranes of bacterial and cancer cells differs markedly from normal human cells, yet the influence of the lipid composition on membrane function is rarely investigated – in part because of the low throughput and high cost of ion transport experiments. Here, we report an increased throughput Workflow to identify biologically relevant, pH-driven ion transport and membrane lysis pathways in vesicle membranes. We developed a set of four assays designed to report on different transport and lysis processes. We validated our assays against a panel of known transporters and produced a stepwise Workflow for the evaluation of libraries of compounds. We applied our Workflow to screen a library (Library 1) of 31 supramolecular, self-associating amphiphiles (SSAs) for transport and lysis activity in a range of vesicles with different lipid compositions, designed to mimic different types of cells, and consequently identified seven promising transporters. Antimicrobial experiments found that six of these promising transporters showed good antimicrobial activity against clinically relevant Staphylococcus aureus strains, highlighting the promise of our Workflow in identifying potential antimicrobial agents. We then applied our Workflow to investigate the ion transport and lysis properties of a second library (Library 2) of SSAs with established antimicrobial and anticancer properties, aiming to provide insight into the biological modes of action.
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d5sc09781a
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Accepted/In Press date: 28 February 2026
e-pub ahead of print date: 3 March 2026
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This journal is © The Royal Society of Chemistry, 2026
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Local EPrints ID: 510652
URI: http://eprints.soton.ac.uk/id/eprint/510652
ISSN: 2041-6520
PURE UUID: cab36105-431f-4cb1-a5fc-4e8283200066
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Date deposited: 15 Apr 2026 16:43
Last modified: 15 Apr 2026 16:43
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Contributors
Author:
Kylie Yang
Author:
Caleb Marsh
Author:
Lisa J. White
Author:
Fergus W. Molyneux
Author:
Thomas L. Allam
Author:
Precious I.A. Popoola
Author:
Olivia B. Keers
Author:
Matthew Rice
Author:
Kira L.F. Hilton
Author:
Hiral A. Kotak
Author:
J. Mark Sutton
Author:
Jose L. Ortega-Roldan
Author:
Charlotte K. Hind
Author:
Jennifer R. Hiscock
Author:
Cally J.E. Haynes
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