Abstract
This study sought to reveal the proteomic profiling of methicillin-resistant Staphylococcus aureus (MRSA)-derived extracellular vesicles (EVs) after exposure to imipenem. The advanced isobaric tags for relative and absolute quantitation (iTRAQ®) proteomic approach were used to analyze the alterations in MRSA-derived EV protein patterns upon exposure to imipenem. A total of 1260 EV proteins were identified and quantified. Among these, 861 differentially expressed exosome proteins (P < 0.05) were found. Multivariate analysis, Gene Ontology (GO) annotation, and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were used to analyze the identified proteins. Enrichment analysis of GO annotations indicated that imipenem primarily regulated the metabolic processes in MRSA. The metabolism of differentially expressed proteins was found to be the most significant in the combined analysis of the KEGG pathway analysis. Based on the results from the STRING analysis, 50S ribosomal protein L16 (RplP) and 30S ribosomal protein S8 (RpsH) were involved in the imipenem-induced MRSA-derived EVs. These results provide vital information on MRSA-derived EVs, increasing our knowledge of the proteome level changes in EVs upon exposure to imipenem. Moreover, these results pave the way for developing novel MRSA treatments.
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The study was funded by the National Natural Science Foundation of China (grant number 81660352).
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In vitro proliferation assay: WJC; data analysis and manuscript writing: WJC and LEM; iTRAQ and MRM data analysis: WJR and RL; protein extraction: WYY and ZCX; bioinformatics analysis: SP; transmission electron microscopy (TEM) and scanning electron microscopy (SEM) analysis: ZXH.
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Supplementary information
Online Resource 1
Global proteomic data in MRSA-derived EVs (XLSX 125 kb)
Online Resource 2
Differentially EVs expressed proteins, volcano plot (A) and histograms (B), between imipenem treated S23 and S23 only (XLSX 163 kb)
Online Resource 3
Gene ontology analysis, including cellular component (A), biological process (B), and molecular function (C), in EVs between imipenem treated S23 and S23 only (XLSX 1442 kb)
Online Resource 4
The KEGG pathway analysis in EVs between imipenem treated S23 and S23 only (XLSX 22 kb)
Online Resource 5
MRM analysis in EVs between imipenem treated S23 and S23 only (XLSX 15 kb)
Online Resource 6
Correlation between MRM and iTRAQ® analysis in EVs between imipenem treated S23 and S23 only (XLSX 17 kb)
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Wang, J., Wang, J., Wang, Y. et al. iTRAQ®-based quantitative proteomics reveals the proteomic profiling of methicillin-resistant Staphylococcus aureus-derived extracellular vesicles after exposure to imipenem. Folia Microbiol 66, 221–230 (2021). https://doi.org/10.1007/s12223-020-00836-y
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DOI: https://doi.org/10.1007/s12223-020-00836-y