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A label-free approach for relative spatial quantitation of c-di-GMP in microbial biofilms

Microbial biofilms represent an important lifestyle for bacteria and are dynamic three dimensional structures. Cyclic dimeric guanosine monophosphate (c-di-GMP) is a ubiquitous signaling molecule that is known to be tightly regulated with biofilm processes. While measurements of global levels of c-d...

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Autores principales: McCaughey, Catherine S., Trebino, Michael A., McAtamney, Allyson, Isenberg, Ruth, Mandel, Mark J., Yildiz, Fitnat H., Sanchez, Laura M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Cold Spring Harbor Laboratory 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10592747/
https://www.ncbi.nlm.nih.gov/pubmed/37873360
http://dx.doi.org/10.1101/2023.10.10.561783
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author McCaughey, Catherine S.
Trebino, Michael A.
McAtamney, Allyson
Isenberg, Ruth
Mandel, Mark J.
Yildiz, Fitnat H.
Sanchez, Laura M.
author_facet McCaughey, Catherine S.
Trebino, Michael A.
McAtamney, Allyson
Isenberg, Ruth
Mandel, Mark J.
Yildiz, Fitnat H.
Sanchez, Laura M.
author_sort McCaughey, Catherine S.
collection PubMed
description Microbial biofilms represent an important lifestyle for bacteria and are dynamic three dimensional structures. Cyclic dimeric guanosine monophosphate (c-di-GMP) is a ubiquitous signaling molecule that is known to be tightly regulated with biofilm processes. While measurements of global levels of c-di-GMP have proven valuable towards understanding the genetic control of c-di-GMP production, there is a need for tools to observe the local changes of c-di-GMP production in biofilm processes. We have developed a label-free method for the direct detection of c-di-GMP in microbial colony biofilms using matrix-assisted laser desorption ionization mass spectrometry imaging (MALDI-MSI). We applied this method to the enteric pathogen Vibrio cholerae, the marine symbiont V. fischeri, and the opportunistic pathogen Pseudomonas aeruginosa PA14 and detected spatial and temporal changes in c-di-GMP signal that accompanied genetic alterations in factors that synthesize and degrade the compound. We further demonstrated how this method can be simultaneously applied to detect additional metabolites of interest in a single experiment.
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spelling pubmed-105927472023-10-24 A label-free approach for relative spatial quantitation of c-di-GMP in microbial biofilms McCaughey, Catherine S. Trebino, Michael A. McAtamney, Allyson Isenberg, Ruth Mandel, Mark J. Yildiz, Fitnat H. Sanchez, Laura M. bioRxiv Article Microbial biofilms represent an important lifestyle for bacteria and are dynamic three dimensional structures. Cyclic dimeric guanosine monophosphate (c-di-GMP) is a ubiquitous signaling molecule that is known to be tightly regulated with biofilm processes. While measurements of global levels of c-di-GMP have proven valuable towards understanding the genetic control of c-di-GMP production, there is a need for tools to observe the local changes of c-di-GMP production in biofilm processes. We have developed a label-free method for the direct detection of c-di-GMP in microbial colony biofilms using matrix-assisted laser desorption ionization mass spectrometry imaging (MALDI-MSI). We applied this method to the enteric pathogen Vibrio cholerae, the marine symbiont V. fischeri, and the opportunistic pathogen Pseudomonas aeruginosa PA14 and detected spatial and temporal changes in c-di-GMP signal that accompanied genetic alterations in factors that synthesize and degrade the compound. We further demonstrated how this method can be simultaneously applied to detect additional metabolites of interest in a single experiment. Cold Spring Harbor Laboratory 2023-10-10 /pmc/articles/PMC10592747/ /pubmed/37873360 http://dx.doi.org/10.1101/2023.10.10.561783 Text en https://creativecommons.org/licenses/by-nc/4.0/This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License (https://creativecommons.org/licenses/by-nc/4.0/) , which allows reusers to distribute, remix, adapt, and build upon the material in any medium or format for noncommercial purposes only, and only so long as attribution is given to the creator.
spellingShingle Article
McCaughey, Catherine S.
Trebino, Michael A.
McAtamney, Allyson
Isenberg, Ruth
Mandel, Mark J.
Yildiz, Fitnat H.
Sanchez, Laura M.
A label-free approach for relative spatial quantitation of c-di-GMP in microbial biofilms
title A label-free approach for relative spatial quantitation of c-di-GMP in microbial biofilms
title_full A label-free approach for relative spatial quantitation of c-di-GMP in microbial biofilms
title_fullStr A label-free approach for relative spatial quantitation of c-di-GMP in microbial biofilms
title_full_unstemmed A label-free approach for relative spatial quantitation of c-di-GMP in microbial biofilms
title_short A label-free approach for relative spatial quantitation of c-di-GMP in microbial biofilms
title_sort label-free approach for relative spatial quantitation of c-di-gmp in microbial biofilms
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10592747/
https://www.ncbi.nlm.nih.gov/pubmed/37873360
http://dx.doi.org/10.1101/2023.10.10.561783
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