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Analysis of the influence of cyclo (L-phenylalanine-L-proline) on the proteome of Staphylococcus aureus using iTRAQ

  • Duiyuan AiEmail author
  • Weibing Zhang
  • Jianmin Yun
  • Yingying Cao
Original Article
  • 4 Downloads

Abstract

Purpose

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).

Methods

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.

Results

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.

Conclusion

cFP reduced AgrA and the expression of some exotoxins but increased Rot and SarR expression.

Keywords

Cyclic dipeptide Quorum sensing Two-component regulatory systems Signal pathway Pathogenicity 

Notes

Acknowledgments

We thank Guangzhou Sagene Biotech Co., Ltd. for help implementing the iTRAQ proteomic experiment.

Funding

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

None.

Informed consent

None.

Supplementary material

13213_2019_1508_MOESM1_ESM.xls (886 kb)
Table S1 Protein identification information. A total of 1296 Staphylococcus aureus CGMCC 1.1861 proteins were identified by Mascot searches and matched peptide information. (XLS) (XLS 885 kb)
13213_2019_1508_MOESM2_ESM.xls (208 kb)
Table S2 Protein quantification information. A total of 1153 Staphylococcus aureus CGMCC 1.1861 proteins were quantified and annotated by KEGG and GO. (XLS) (XLS 207 kb)
13213_2019_1508_MOESM3_ESM.xls (20 kb)
Table S3 Significantly differentially expressed proteins. A total of 84 DEPs of Staphylococcus aureus CGMCC 1.1861 were quantified. Proteins with a fold change >1.2 or < 0.83 and an unadjusted significance level of P < 0.05 were considered differentially expressed between control and treatment groups. (XLS) (XLS 20 kb)
13213_2019_1508_MOESM4_ESM.xls (25 kb)
Table S4 Pathway annotation. A total of 522 Staphylococcus aureus CGMCC 1.1861 proteins were classified into 98 categories based on KEGG metabolic pathways. (XLS) (XLS 24 kb)
13213_2019_1508_MOESM5_ESM.xls (121 kb)
Table S5 COG classification. A total of 1038 Staphylococcus aureus CGMCC 1.1861 proteins were classified into 20 groups based on COG categories. (XLS) (XLS 120 kb)
13213_2019_1508_MOESM6_ESM.xls (6 kb)
Table S6 Enriched pathways ofS. aureusproteins influenced by cFP. A total of 48 Staphylococcus aureus CGMCC 1.1861 pathways were analyzed using the pathway enrichment method. Proteins with P < 0.05 were considered differentially expressed between control and treatment groups. (XLS) (XLS 5 kb)
13213_2019_1508_Fig5_ESM.png (6.2 mb)
Figure S7

Proteins identified as signal transduction TCRS by KEGG analysis. The red rectangular box represents identified Staphylococcus aureus CGMCC 1.1861 proteins that were classified as two-component regulatory systems (TCRS). (TIF) (PNG 6330 kb)

13213_2019_1508_MOESM7_ESM.tif (559 kb)
High resolution image (TIF 559 kb)
13213_2019_1508_Fig6_ESM.png (409 kb)
Figure S8

All pathways enriched forS. aureusproteins influenced by cFP. The first line “class” indicates the type of pathway and the second line “CK-VS-cFP” indicates their significance level. (TIF) (PNG 408 kb)

13213_2019_1508_MOESM8_ESM.tif (1.1 mb)
High resolution image (TIF 1102 kb)

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Copyright information

© Università degli studi di Milano 2019

Authors and Affiliations

  1. 1.College of Food Science and EngineeringGansu Agricultural UniversityLanzhouPeople’s Republic of China
  2. 2.Food Science and EngineeringGansu Agricultural UniversityLanzhou cityChina

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