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

Autor:	McCaughey CS; Department of Chemistry and Biochemistry, University of California, Santa Cruz, Santa Cruz, CA 95064., Trebino MA; Department of Microbiology and Environmental Toxicology, University of California, Santa Cruz, Santa Cruz, CA 95064., McAtamney A; Department of Chemistry and Biochemistry, University of California, Santa Cruz, Santa Cruz, CA 95064., Isenberg R; Department of Medical Microbiology and Immunology, University of Wisconsin-Madison, Madison, WI 53706.; Microbiology Doctoral Training Program, University of Wisconsin-Madison, Madison, WI 53706.; Current Address: Department of Microbiology and Immunology, University of Minnesota Medical School, Minneapolis, MN 55455., Mandel MJ; Department of Medical Microbiology and Immunology, University of Wisconsin-Madison, Madison, WI 53706.; Microbiology Doctoral Training Program, University of Wisconsin-Madison, Madison, WI 53706., Yildiz FH; Department of Microbiology and Environmental Toxicology, University of California, Santa Cruz, Santa Cruz, CA 95064., Sanchez LM; Department of Chemistry and Biochemistry, University of California, Santa Cruz, Santa Cruz, CA 95064.
Jazyk:	angličtina
Zdroj:	BioRxiv : the preprint server for biology [bioRxiv] 2023 Oct 10. Date of Electronic Publication: 2023 Oct 10.
DOI:	10.1101/2023.10.10.561783
Abstrakt:	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.
Databáze:	MEDLINE
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