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Coupling MALDI-TOF mass spectrometry protein and specialized metabolite analyses to rapidly discriminate bacterial function

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dc.contributor Háskóli Íslands (HÍ)
dc.contributor University of Iceland (UI)
dc.contributor.author Clark, Chase M.
dc.contributor.author Costa, Maria Sofia
dc.contributor.author Sanchez, Laura M.
dc.contributor.author Murphy, Brian T.
dc.date.accessioned 2020-01-30T17:16:32Z
dc.date.available 2020-01-30T17:16:32Z
dc.date.issued 2018-04-23
dc.identifier.citation Clark, C., Costa, M., Sanchez, L., & Murphy, B. (2018). Coupling MALDI-TOF mass spectrometry protein and specialized metabolite analyses to rapidly discriminate bacterial function. Proceedings of the National Academy of Sciences of the United States of America, 115(19), 4981-4986.
dc.identifier.issn 1091-6490
dc.identifier.issn 0027-8424 (eISSN)
dc.identifier.uri https://hdl.handle.net/20.500.11815/1491
dc.description Publisher's version (útgefin grein).
dc.description.abstract For decades, researchers have lacked the ability to rapidly correlate microbial identity with bacterial metabolism. Since specialized metabolites are critical to bacterial function and survival in the environment, we designed a data acquisition and bioinformatics technique (IDBac) that utilizes in situ matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) to analyze protein and specialized metabolite spectra recorded from single bacterial colonies picked from agar plates. We demonstrated the power of our approach by discriminating between two Bacillus subtilis strains in <30 min solely on the basis of their differential ability to produce cyclic peptide antibiotics surfactin and plipastatin, caused by a single frameshift mutation. Next, we used IDBac to detect subtle intraspecies differences in the production of metal scavenging acyl-desferrioxamines in a group of eight freshwater Micromonospora isolates that share >99% sequence similarity in the 16S rRNA gene. Finally, we used IDBac to simultaneously extract protein and specialized metabolite MS profiles from unidentified Lake Michigan sponge-associated bacteria isolated from an agar plate. In just 3 h, we created hierarchical protein MS groupings of 11 environmental isolates (10 MS replicates each, for a total of 110 spectra) that accurately mirrored phylogenetic groupings. We further distinguished isolates within these groupings, which share nearly identical 16S rRNA gene sequence identity, based on interspecies and intraspecies differences in specialized metabolite production. IDBac is an attempt to couple in situ MS analyses of protein content and specialized metabolite production to allow for facile discrimination of closely related bacterial colonies.
dc.description.sponsorship We thank the following contributors: Russel Cuhel and crew of R. V. Neeskay (University of Milwaukee Wisconsin) for assistance with sediment collection; Keith Jung for collection of the freshwater sponge; Dr. Amanda Bulman (Bruker) for assistance with MALDI-TOF MS protein acquisition parameters; Dr. Terry Moore and Dr. Atul Jain for recrystallizing alpha-cyano-4-hydroxycinnamic acid matrix (CHCA); and Dr. Darrell McCaslin (University of Wisconsin Madison) for testing our method on legacy instruments. This work was supported by National Institute of Child Health and Human Development Grant K12HD055892; the National Institutes of Health Office of Research on Women’s Health; National Geographic Grant CP-044R17; Icelandic Research Fund Grant 152336-051; and University of Illinois at Chicago startup funds.
dc.format.extent 4981-4986
dc.language.iso en
dc.publisher Proceedings of the National Academy of Sciences
dc.relation.ispartofseries Proceedings of the National Academy of Sciences;115(19)
dc.rights info:eu-repo/semantics/openAccess
dc.subject Bioinformatics
dc.subject Mass spectrometry
dc.subject Metabolomics
dc.subject Natural products
dc.subject Specialized metabolites
dc.subject Náttúruefni (lyfjafræði)
dc.subject Litrófsgreining
dc.subject Efnahvörf
dc.title Coupling MALDI-TOF mass spectrometry protein and specialized metabolite analyses to rapidly discriminate bacterial function
dc.type info:eu-repo/semantics/article
dcterms.license This open access article is distributed under Creative Commons Attribution-NonCommercialNoDerivatives License 4.0 (CC BY-NC-ND).
dc.description.version Peer Reviewed
dc.identifier.journal Proceedings of the National Academy of Sciences
dc.identifier.doi 10.1073/pnas.1801247115
dc.contributor.department Faculty of Pharmaceutical Sciences (UI)
dc.contributor.department Lyfjafræðideild (HÍ)
dc.contributor.school Heilbrigðisvísindasvið (HÍ)
dc.contributor.school School of Health Sciences (UI)

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