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Comparative Proteomic Analysis of Adhesion/Invasion Related Proteins in Cronobacter sakazakii Based on
Ping Li1, Xuan Dong1, Xiao-Yi Wang1
1State Key Laboratory of Food Nutrition and Safety, Tianjin University of Science and Technology, Tianjin, China.
Insights
This study identified key proteins in Cronobacter sakazakii linked to its ability to adhere to and invade host cells. These findings offer insights into the virulence mechanisms of this foodborne pathogen.
Area of Science:
- Microbiology
- Food Safety
- Proteomics
Background:
- Cronobacter sakazakii is a foodborne pathogen causing severe infant illnesses.
- The virulence factors and pathogenicity mechanisms of C. sakazakii are not fully understood.
Purpose of the Study:
- To identify differentially expressed proteins associated with the adhesion and invasion capabilities of C. sakazakii.
- To investigate potential virulence determinants of C. sakazakii using proteomic analysis.
Main Methods:
- Multilocus sequence typing (MLST) was used to characterize C. sakazakii strains.
- Data-independent acquisition (DIA) proteomic analysis was employed to identify differentially expressed proteins.
- Real-time quantitative PCR (qPCR) was used to validate gene expression patterns.
Main Results:
- A total of 2,203 proteins were identified, with 210 showing differential expression between strongly and weakly adhesive/invasive strains.
- Highly expressed proteins in the strongly adhesive/invasive strain were associated with flagellar assembly, lipopolysaccharide synthesis, LuxS/AI-2 signaling, energy metabolism, and iron-sulfur clusters.
- Proteomic findings were consistent with transcriptional analysis.
Conclusions:
- This study provides the first use of DIA proteomic analysis to explore adhesion/invasion factors in C. sakazakii.
- Identified proteins offer a valuable reference for understanding C. sakazakii pathogenicity mechanisms.
Abstract:
Cronobacter sakazakii is foodborne pathogen that causes serious illnesses such as necrotizing enterocolitis, meningitis and septicemia in infants. However, the virulence determinants and mechanisms of pathogenicity of these species remain unclear. In this study, multilocus sequence typing (MLST) was performed on 34 C. sakazakii strains and two strains with the same sequence type (ST) but distinct adhesion/invasion capabilities were selected for identification of differentially expressed proteins using data-independent acquisition (DIA) proteomic analysis. A total of 2,203 proteins were identified and quantified. Among these proteins, 210 exhibited differential expression patterns with abundance ratios ≥3 or ≤0.33 and P values ≤0.05. Among these 210 proteins, 67 were expressed higher, and 143 were expressed lower in C. sakazakii SAKA80220 (strongly adhesive/invasive strain) compared with C. sakazakii SAKA80221 (weakly adhesive/invasive strain). Based on a detailed analysis of the differentially expressed proteins, the highly expressed genes involved in flagellar assembly, lipopolysaccharide synthesis, LuxS/AI-2, energy metabolic pathways and iron-sulfur cluster may be associated with the adhesion/invasion capability of C. sakazakii. To verify the accuracy of the proteomic results, real-time qPCR was used to analyze the expression patterns of some genes at the transcriptional level, and consistent results were observed. This study, for the first time, used DIA proteomic to investigate potential adhesion/invasion related factors as a useful reference for further studies on the pathogenic mechanism of C. sakazakii.
