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Biomimetic Materials to Characterize Bacteria-host Interactions
Bacterial attachment to host cells is one of the earliest events during bacterial colonization of host tissues and thus a key step during infection. The biochemical and functional characterization of adhesins mediating these initial bacteria-host interactions is often compromised by the presence of...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MyJove Corporation
2015
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4692734/ https://www.ncbi.nlm.nih.gov/pubmed/26650964 http://dx.doi.org/10.3791/53400 |
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author | Stones, Daniel H. Al-Saedi, Fitua Vaz, Diana Perez-Soto, Nicolas Krachler, Anne M. |
author_facet | Stones, Daniel H. Al-Saedi, Fitua Vaz, Diana Perez-Soto, Nicolas Krachler, Anne M. |
author_sort | Stones, Daniel H. |
collection | PubMed |
description | Bacterial attachment to host cells is one of the earliest events during bacterial colonization of host tissues and thus a key step during infection. The biochemical and functional characterization of adhesins mediating these initial bacteria-host interactions is often compromised by the presence of other bacterial factors, such as cell wall components or secreted molecules, which interfere with the analysis. This protocol describes the production and use of biomimetic materials, consisting of pure recombinant adhesins chemically coupled to commercially available, functionalized polystyrene beads, which have been used successfully to dissect the biochemical and functional interactions between individual bacterial adhesins and host cell receptors. Protocols for different coupling chemistries, allowing directional immobilization of recombinant adhesins on polymer scaffolds, and for assessment of the coupling efficiency of the resulting “bacteriomimetic” materials are also discussed. We further describe how these materials can be used as a tool to inhibit pathogen mediated cytotoxicity and discuss scope, limitations and further applications of this approach in studying bacterial - host interactions. |
format | Online Article Text |
id | pubmed-4692734 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | MyJove Corporation |
record_format | MEDLINE/PubMed |
spelling | pubmed-46927342016-01-07 Biomimetic Materials to Characterize Bacteria-host Interactions Stones, Daniel H. Al-Saedi, Fitua Vaz, Diana Perez-Soto, Nicolas Krachler, Anne M. J Vis Exp Infection Bacterial attachment to host cells is one of the earliest events during bacterial colonization of host tissues and thus a key step during infection. The biochemical and functional characterization of adhesins mediating these initial bacteria-host interactions is often compromised by the presence of other bacterial factors, such as cell wall components or secreted molecules, which interfere with the analysis. This protocol describes the production and use of biomimetic materials, consisting of pure recombinant adhesins chemically coupled to commercially available, functionalized polystyrene beads, which have been used successfully to dissect the biochemical and functional interactions between individual bacterial adhesins and host cell receptors. Protocols for different coupling chemistries, allowing directional immobilization of recombinant adhesins on polymer scaffolds, and for assessment of the coupling efficiency of the resulting “bacteriomimetic” materials are also discussed. We further describe how these materials can be used as a tool to inhibit pathogen mediated cytotoxicity and discuss scope, limitations and further applications of this approach in studying bacterial - host interactions. MyJove Corporation 2015-11-16 /pmc/articles/PMC4692734/ /pubmed/26650964 http://dx.doi.org/10.3791/53400 Text en Copyright © 2015, Journal of Visualized Experiments http://creativecommons.org/licenses/by-nc-nd/3.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported License. To view a copy of this license, visithttp://creativecommons.org/licenses/by-nc-nd/3.0/ |
spellingShingle | Infection Stones, Daniel H. Al-Saedi, Fitua Vaz, Diana Perez-Soto, Nicolas Krachler, Anne M. Biomimetic Materials to Characterize Bacteria-host Interactions |
title | Biomimetic Materials to Characterize Bacteria-host Interactions |
title_full | Biomimetic Materials to Characterize Bacteria-host Interactions |
title_fullStr | Biomimetic Materials to Characterize Bacteria-host Interactions |
title_full_unstemmed | Biomimetic Materials to Characterize Bacteria-host Interactions |
title_short | Biomimetic Materials to Characterize Bacteria-host Interactions |
title_sort | biomimetic materials to characterize bacteria-host interactions |
topic | Infection |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4692734/ https://www.ncbi.nlm.nih.gov/pubmed/26650964 http://dx.doi.org/10.3791/53400 |
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