Analysis of the influence of cyclo (L-phenylalanine-L-proline) on the proteome of Staphylococcus aureus using iTRAQ
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Cyclo (L-phenylalanine-L-proline) (cFP) is an extracellular quorum sensing (QS) signal molecule that coordinates communication between Gram-negative bacteria. Some studies have also reported QS in Gram-positive bacteria. However, the effect of cFP on Gram-positive bacteria remains unknown. Therefore, an isobaric tags for relative and absolute quantitation (iTRAQ) proteomic experiment were designed to elucidate whether cFP influences protein expression in Staphylococcus aureus (S. aureus).
The iTRAQ proteomics method was used to analyze untreated (control) and S. aureus treated with cFP for 12 h. Samples were then processed by liquid-phase tandem mass spectrometry (LC-MS/MS) and analyzed using bioinformatics tools.
The results identified 1296 proteins from the S. aureus CGMCC 1.1861 proteome. Twenty-two proteins, including some two-component regulatory systems (TCRS), were associated with signal transduction. Differential expression analysis revealed that only 43 proteins were up-regulated and 41 proteins were down-regulated by cFP. The most significantly different pathways were amino acid metabolism, fatty acid degradation, and metabolism of cofactors and vitamins. Results showed that cFP down-regulated virulence factors, up-regulated lipid and amino acid metabolism, promoted acetylation and phosphorylation, and decreased alcohol dehydrogenase expression. A total of 12 significantly differentially expressed proteins (DEPs) were related to signal transduction. Among them, Rot (Q9RFJ6) and SarR (Q9F0R1), which can inhibit transcription of the Agr system, were up-regulated, whereas virulence factors such as ESAT-6 protein A (Q2G189), phenol soluble modulin (Psm, Q2FZA4), and a peptide ABC transporter permease (Q2G168) were down-regulated. AgrA (Q2FWM4) was down-regulated by cFP in S. aureus.
cFP reduced AgrA and the expression of some exotoxins but increased Rot and SarR expression.
KeywordsCyclic dipeptide Quorum sensing Two-component regulatory systems Signal pathway Pathogenicity
We thank Guangzhou Sagene Biotech Co., Ltd. for help implementing the iTRAQ proteomic experiment.
Research reported in this publication was supported by the National Natural Science Foundation of China (NSFC grant numbers 31460425, 31560442, 31760466, and 31360405).
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
Research involving human participants and/or animals
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