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Probing Interkingdom Signaling Molecules via Liquid Extraction Surface Analysis-Mass Spectrometry

Robertson, Shaun N.; Soukarieh, Fadi; White, Thomas M.; Camara, Miguel; Romero, Manuel; Griffiths, Rian L.


Thomas M. White

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Professor of Molecular Microbiology

Manuel Romero


Previously, metabolites diffused or secreted from microbial samples have been analyzed via liquid chromatography–mass spectrometry (LC–MS) approaches following lengthy extraction protocols. Here, we present a model system for growing biofilms on discs before utilizing rapid and direct surface sampling MS, namely, liquid extraction surface analysis, to study the microbial exometabolome. One of the benefits of this approach is its surface-specific nature, enabling mimicking biofilm formation in a way that the study of planktonic liquid cultures cannot imitate. Even though Pseudomonas aeruginosa (P. aeruginosa), Staphylococcus aureus (S. aureus), and Candida albicans (C. albicans) have been studied previously in isolation, very few studies consider the complexity of the interplay between these pathogens, which are commonly combined causative agents of infection. Our model system provides a route to investigate changes in the exometabolome, such as metabolites that become circulatory in the presence of multiple pathogens. Our results agree with previous reports showing that 2-alkyl-4(1H)-quinolone signal molecules produced by P. aeruginosa are important markers of infection and suggest that methods for monitoring levels of 2-heptyl-4-hydroxyquinoline and 2,4-dihydroxyquinoline, as well as pyocyanin, could be beneficial in the determination of causative agents in interkingdom infection including P. aeruginosa. Furthermore, studying changes in exometabolome metabolites between pqs quorum sensing antagonists in treated and nontreated samples suggests suppression of phenazine production by P. aeruginosa. Hence, our model provides a rapid analytical approach to gaining a mechanistic understanding of bacterial signaling.


Robertson, S. N., Soukarieh, F., White, T. M., Camara, M., Romero, M., & Griffiths, R. L. (2023). Probing Interkingdom Signaling Molecules via Liquid Extraction Surface Analysis-Mass Spectrometry. Analytical Chemistry, 95(11), 5079-5086.

Journal Article Type Article
Acceptance Date Feb 21, 2023
Online Publication Date Mar 7, 2023
Publication Date Mar 21, 2023
Deposit Date Mar 10, 2023
Publicly Available Date Mar 8, 2024
Journal Analytical Chemistry
Print ISSN 0003-2700
Electronic ISSN 1520-6882
Publisher American Chemical Society
Peer Reviewed Peer Reviewed
Volume 95
Issue 11
Pages 5079-5086
Keywords Biofilms, Computer simulations, Infectious diseases, Inhibitors, Molecules
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