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) |