Cargando…
Microspectrometric insights on the uptake of antibiotics at the single bacterial cell level
Bacterial multidrug resistance is a significant health issue. A key challenge, particularly in Gram-negative antibacterial research, is to better understand membrane permeation of antibiotics in clinically relevant bacterial pathogens. Passing through the membrane barrier to reach the required conce...
Autores principales: | , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group
2015
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4675965/ https://www.ncbi.nlm.nih.gov/pubmed/26656111 http://dx.doi.org/10.1038/srep17968 |
_version_ | 1782405082815397888 |
---|---|
author | Cinquin, Bertrand Maigre, Laure Pinet, Elizabeth Chevalier, Jacqueline Stavenger, Robert A. Mills, Scott Réfrégiers, Matthieu Pagès, Jean-Marie |
author_facet | Cinquin, Bertrand Maigre, Laure Pinet, Elizabeth Chevalier, Jacqueline Stavenger, Robert A. Mills, Scott Réfrégiers, Matthieu Pagès, Jean-Marie |
author_sort | Cinquin, Bertrand |
collection | PubMed |
description | Bacterial multidrug resistance is a significant health issue. A key challenge, particularly in Gram-negative antibacterial research, is to better understand membrane permeation of antibiotics in clinically relevant bacterial pathogens. Passing through the membrane barrier to reach the required concentration inside the bacterium is a pivotal step for most antibacterials. Spectrometric methodology has been developed to detect drugs inside bacteria and recent studies have focused on bacterial cell imaging. Ultimately, we seek to use this method to identify pharmacophoric groups which improve penetration, and therefore accumulation, of small-molecule antibiotics inside bacteria. We developed a method to quantify the time scale of antibiotic accumulation in living bacterial cells. Tunable ultraviolet excitation provided by DISCO beamline (synchrotron Soleil) combined with microscopy allows spectroscopic analysis of the antibiotic signal in individual bacterial cells. Robust controls and measurement of the crosstalk between fluorescence channels can provide real time quantification of drug. This technique represents a new method to assay drug translocation inside the cell and therefore incorporate rational drug design to impact antibiotic uptake. |
format | Online Article Text |
id | pubmed-4675965 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | Nature Publishing Group |
record_format | MEDLINE/PubMed |
spelling | pubmed-46759652015-12-16 Microspectrometric insights on the uptake of antibiotics at the single bacterial cell level Cinquin, Bertrand Maigre, Laure Pinet, Elizabeth Chevalier, Jacqueline Stavenger, Robert A. Mills, Scott Réfrégiers, Matthieu Pagès, Jean-Marie Sci Rep Article Bacterial multidrug resistance is a significant health issue. A key challenge, particularly in Gram-negative antibacterial research, is to better understand membrane permeation of antibiotics in clinically relevant bacterial pathogens. Passing through the membrane barrier to reach the required concentration inside the bacterium is a pivotal step for most antibacterials. Spectrometric methodology has been developed to detect drugs inside bacteria and recent studies have focused on bacterial cell imaging. Ultimately, we seek to use this method to identify pharmacophoric groups which improve penetration, and therefore accumulation, of small-molecule antibiotics inside bacteria. We developed a method to quantify the time scale of antibiotic accumulation in living bacterial cells. Tunable ultraviolet excitation provided by DISCO beamline (synchrotron Soleil) combined with microscopy allows spectroscopic analysis of the antibiotic signal in individual bacterial cells. Robust controls and measurement of the crosstalk between fluorescence channels can provide real time quantification of drug. This technique represents a new method to assay drug translocation inside the cell and therefore incorporate rational drug design to impact antibiotic uptake. Nature Publishing Group 2015-12-11 /pmc/articles/PMC4675965/ /pubmed/26656111 http://dx.doi.org/10.1038/srep17968 Text en Copyright © 2015, Macmillan Publishers Limited http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ |
spellingShingle | Article Cinquin, Bertrand Maigre, Laure Pinet, Elizabeth Chevalier, Jacqueline Stavenger, Robert A. Mills, Scott Réfrégiers, Matthieu Pagès, Jean-Marie Microspectrometric insights on the uptake of antibiotics at the single bacterial cell level |
title | Microspectrometric insights on the uptake of antibiotics at the single bacterial cell level |
title_full | Microspectrometric insights on the uptake of antibiotics at the single bacterial cell level |
title_fullStr | Microspectrometric insights on the uptake of antibiotics at the single bacterial cell level |
title_full_unstemmed | Microspectrometric insights on the uptake of antibiotics at the single bacterial cell level |
title_short | Microspectrometric insights on the uptake of antibiotics at the single bacterial cell level |
title_sort | microspectrometric insights on the uptake of antibiotics at the single bacterial cell level |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4675965/ https://www.ncbi.nlm.nih.gov/pubmed/26656111 http://dx.doi.org/10.1038/srep17968 |
work_keys_str_mv | AT cinquinbertrand microspectrometricinsightsontheuptakeofantibioticsatthesinglebacterialcelllevel AT maigrelaure microspectrometricinsightsontheuptakeofantibioticsatthesinglebacterialcelllevel AT pinetelizabeth microspectrometricinsightsontheuptakeofantibioticsatthesinglebacterialcelllevel AT chevalierjacqueline microspectrometricinsightsontheuptakeofantibioticsatthesinglebacterialcelllevel AT stavengerroberta microspectrometricinsightsontheuptakeofantibioticsatthesinglebacterialcelllevel AT millsscott microspectrometricinsightsontheuptakeofantibioticsatthesinglebacterialcelllevel AT refregiersmatthieu microspectrometricinsightsontheuptakeofantibioticsatthesinglebacterialcelllevel AT pagesjeanmarie microspectrometricinsightsontheuptakeofantibioticsatthesinglebacterialcelllevel |