Fluoroquinolone resistant Salmonella proteomics analysis based on tandem mass tag and Parallel reaction monitoring techniques

Abstract:

[Background] As Salmonella resistance to fluoroquinolones has increased gradually, it is very urgent and important to study the mechanism of drug resistance. Proteomics analysis will provide new targets and directions for the study on resistance mechanism in Salmonella. [Objective] Proteomic analysis of Salmonella typhimurium was performed before and after induction, in order to lay the foundation for further studies on the mechanism in Salmonella resistance. [Methods] Ciprofloxacin was used to induce Salmonella typhimurium ATCC13311 to obtain resistance, screening and bioinformatics analysis of differential proteins were performed by using tandem mass tag (TMT). Besides, fifteen differential proteins were selected for (parallel reaction monitoring PRM) target verification. [Results] The results showed that a total of 318 differentially expressed proteins were screened, among which 159 proteins were up-regulated and 159 proteins were down-regulated. The KEGG pathways involved in these proteins mainly including bacterial chemotaxis, ABC transporters, two-component systems, etc; PRM was successfully quantitated to 13 validated proteins and the trend was consistent with TMT. [Conclusion] In this study, the proteomics analysis of Salmonella typhimurium parental strain and ciprofloxacin-resistant strain was conducted by TMT quantitatively combined with PRM target verification, screening out a variety of differential proteins and metabolic pathways, including efflux pump related proteins, outer membrane proteins, two-component related proteins and pathways, bacteria chemotaxis related proteins and pathways, etc. The study laid a certain foundation for further studies on the mechanism of fluoroquinolone resistance in Salmonella.

Key words:Salmonella, Drug resistance, Proteomics, Tandem mass tag, Parallel reaction monitoring