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• W2897465175 endingPage "3996" @default.
• W2897465175 startingPage "3984" @default.
• W2897465175 abstract "Bacterial persisters, a dormant and multidrug tolerant subpopulation that are able to resuscitate after antibiotic treatment, have recently received considerable attention as a major cause of relapse of various infectious diseases in the clinic. However, because of their low abundance and inherent transience, it is extremely difficult to study them by proteomics. Here we developed a magnetic-beads-based separation approach to enrich Escherichia coli persisters and then subjected them to shotgun proteomics. Rifampin pretreatment was employed to increase persister formation, and the resulting cells were exposed to a high concentration of ampicillin (10× MIC) to remove nonpersisters. The survivors were analyzed by spectral counting-based quantitative proteomics. On average, 710 proteins were identified at a false discovery rate of 0.01 for enriched E. coli persisters. By spectral counting-based quantification, 105 proteins (70 down-regulated, 35 up-regulated) were shown to be differentially expressed compared with normal cells. A comparison of the differentially expressed proteins between the magnetic beads-enriched persisters and nonenriched persisters (a mixture of persisters and intact dead cells) shows only around half (∼58%) overlap and different protein–protein interaction networks. This suggest that persister enrichment is important to eliminate the cumulative effect of dead cells that will obscure the proteome of persisters. As expected, proteins involved in carbohydrate metabolism, fatty acid and amino acid biosynthesis, and bacterial chemotaxis were found to be down-regulated in the persisters. Interestingly, membrane proteins including some transport proteins were up-regulated, indicating that they might be important for the drug tolerance of persisters. Knockout of the pal gene expressing peptidoglycan-associated lipoprotein, one of the most up-regulated proteins detected in persisters, led to 10-fold reduced persister formation under ampicillin treatment." @default.
• W2897465175 created "2018-10-26" @default.
• W2897465175 creator A5045310678 @default.
• W2897465175 creator A5068077119 @default.
• W2897465175 creator A5085021194 @default.
• W2897465175 date "2018-10-11" @default.
• W2897465175 modified "2023-09-30" @default.
• W2897465175 title "Specific Enrichment and Proteomics Analysis of <i>Escherichia coli</i> Persisters from Rifampin Pretreatment" @default.
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• W2897465175 doi "https://doi.org/10.1021/acs.jproteome.8b00625" @default.
• W2897465175 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/30336045" @default.
• W2897465175 hasPublicationYear "2018" @default.
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