iTRAQ®-based quantitative proteomics reveals the proteomic profiling of methicillin-resistant Staphylococcus aureus-derived extracellular vesicles after exposure to imipenem

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.

Exosomes: Potential Disease Biomarkers and New Therapeutic Targets

“…Complete identification of exosomal protein cargo can be achieved with a "global discovery" approach using multidimensional fractionation strategies followed by high-resolution mass spectrometric (MS) [326], which could include a quantitative cargo analysis if desired. This quantification can be performed with the Stable Isotope Labeling by Amino acids in Cell culture (SILAC) or Isobaric Tag for Relative and Absolute Quantitation (iTRAQ) methods, involving the incorporation of a stable heavy isotope-labeled amino acid into the peptides of interests [327,328]. However, these are expensive protocols, and the peptide of interest is not always known.…”

Impact of Environmental Conditions on the Protein Content of Staphylococcus aureus and Its Derived Extracellular Vesicles

“…Concerning EV composition, a recent study by Briaud et al demonstrated that temperature variations affect the EV protein and RNA cargo quantitatively and qualitatively, which is also reflected in different EV cytolytic activities towards host cells [40]. Culture media, growth phase, and antibiotic treatment were also shown to alter S. aureus EV content [39,[43][44][45]. However, studies comparing the impact of different environmental conditions on both S. aureus producing cells and its derived EVs are still lacking.…”

Carrier-Free Binary Self-Assembled Nanomedicines Originated from Traditional Herb Medicine with Multifunction to Accelerate MRSA-Infected Wound Healing by Antibacterial, Anti-Inflammation and Promoting Angiogenesis

“…These DEGs were all related to the synthesis of ribosomal proteins, which in turn affected the translation machinery and led to bacterial death. [48][49][50][51] Both of them mRNA expression of DEGs were down regulated, and the gene regulation effect of NPs was more prominent than that of BBR, resulting in the antibacterial effect of NPs was far superior to BBR. Besides, most of these DEGs could also be found in GA group.…”