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An Efficient Way to Screen Inhibitors of Energy-Coupling Factor (ECF) Transporters in a Bacterial Uptake Assay
Herein, we report a novel whole-cell screening assay using Lactobacillus casei as a model microorganism to identify inhibitors of energy-coupling factor (ECF) transporters. This promising and underexplored target may have important pharmacological potential through modulation of vitamin homeostasis...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8910649/ https://www.ncbi.nlm.nih.gov/pubmed/35269783 http://dx.doi.org/10.3390/ijms23052637 |
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author | Bousis, Spyridon Winkler, Steffen Haupenthal, Jörg Fulco, Francesco Diamanti, Eleonora Hirsch, Anna K. H. |
author_facet | Bousis, Spyridon Winkler, Steffen Haupenthal, Jörg Fulco, Francesco Diamanti, Eleonora Hirsch, Anna K. H. |
author_sort | Bousis, Spyridon |
collection | PubMed |
description | Herein, we report a novel whole-cell screening assay using Lactobacillus casei as a model microorganism to identify inhibitors of energy-coupling factor (ECF) transporters. This promising and underexplored target may have important pharmacological potential through modulation of vitamin homeostasis in bacteria and, importantly, it is absent in humans. The assay represents an alternative, cost-effective and fast solution to demonstrate the direct involvement of these membrane transporters in a native biological environment rather than using a low-throughput in vitro assay employing reconstituted proteins in a membrane bilayer system. Based on this new whole-cell screening approach, we demonstrated the optimization of a weak hit compound (2) into a small molecule (3) with improved in vitro and whole-cell activities. This study opens the possibility to quickly identify novel inhibitors of ECF transporters and optimize them based on structure–activity relationships. |
format | Online Article Text |
id | pubmed-8910649 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-89106492022-03-11 An Efficient Way to Screen Inhibitors of Energy-Coupling Factor (ECF) Transporters in a Bacterial Uptake Assay Bousis, Spyridon Winkler, Steffen Haupenthal, Jörg Fulco, Francesco Diamanti, Eleonora Hirsch, Anna K. H. Int J Mol Sci Article Herein, we report a novel whole-cell screening assay using Lactobacillus casei as a model microorganism to identify inhibitors of energy-coupling factor (ECF) transporters. This promising and underexplored target may have important pharmacological potential through modulation of vitamin homeostasis in bacteria and, importantly, it is absent in humans. The assay represents an alternative, cost-effective and fast solution to demonstrate the direct involvement of these membrane transporters in a native biological environment rather than using a low-throughput in vitro assay employing reconstituted proteins in a membrane bilayer system. Based on this new whole-cell screening approach, we demonstrated the optimization of a weak hit compound (2) into a small molecule (3) with improved in vitro and whole-cell activities. This study opens the possibility to quickly identify novel inhibitors of ECF transporters and optimize them based on structure–activity relationships. MDPI 2022-02-27 /pmc/articles/PMC8910649/ /pubmed/35269783 http://dx.doi.org/10.3390/ijms23052637 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Bousis, Spyridon Winkler, Steffen Haupenthal, Jörg Fulco, Francesco Diamanti, Eleonora Hirsch, Anna K. H. An Efficient Way to Screen Inhibitors of Energy-Coupling Factor (ECF) Transporters in a Bacterial Uptake Assay |
title | An Efficient Way to Screen Inhibitors of Energy-Coupling Factor (ECF) Transporters in a Bacterial Uptake Assay |
title_full | An Efficient Way to Screen Inhibitors of Energy-Coupling Factor (ECF) Transporters in a Bacterial Uptake Assay |
title_fullStr | An Efficient Way to Screen Inhibitors of Energy-Coupling Factor (ECF) Transporters in a Bacterial Uptake Assay |
title_full_unstemmed | An Efficient Way to Screen Inhibitors of Energy-Coupling Factor (ECF) Transporters in a Bacterial Uptake Assay |
title_short | An Efficient Way to Screen Inhibitors of Energy-Coupling Factor (ECF) Transporters in a Bacterial Uptake Assay |
title_sort | efficient way to screen inhibitors of energy-coupling factor (ecf) transporters in a bacterial uptake assay |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8910649/ https://www.ncbi.nlm.nih.gov/pubmed/35269783 http://dx.doi.org/10.3390/ijms23052637 |
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