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Published on: April 25, 2015
Proteomic analysis of the copper resistance of Streptococcus pneumoniae
Zhong Guo1, Junlong Han, Xiao-Yan Yang
1Key Laboratory of Functional Protein Research of Guangdong Higher Education Institutes, Institute of Life and Health Engineering, College of Life Science and Technology, Jinan University, Guangzhou 510632, China. tqyhe@jnu.edu.cn tsunxs@jnu.edu.cn.
Abstract:
Streptococcus pneumoniae is a Gram-positive bacterial pathogen causing a variety of diseases, including otitis media, bacteraemia and meningitis. Although copper is an essential trace metal for bacterial growth, high intracellular levels of free-copper are toxic. Copper resistance has emerged as an important virulence determinant of microbial pathogens. In this study, we determined the minimum inhibition concentration of copper for the growth inhibition of S. pneumoniae. Two-dimensional-electrophoresis coupled with mass spectrometry was applied to identify proteins involved in copper resistance of S. pneumoniae. In total, forty-four proteins with more than 1.5-fold alteration in expression (p < 0.05) were identified. Quantitative reverse transcription PCR was used to confirm the proteomic results. Bioinformatics analysis showed that the differentially expressed proteins were mainly involved in the cell wall biosynthesis, protein biosynthesis, purine biosynthesis, pyrimidine biosynthesis, primary metabolic process, and the nitrogen compound metabolic process. Many up-regulated proteins in response to the copper treatment directly or indirectly participated in the cell wall biosynthesis, indicating that the cell wall is a critical determinant in copper resistance of S. pneumoniae.
Insights
This study investigated copper resistance in Streptococcus pneumoniae, a common bacterial pathogen. Results show copper impacts cell wall biosynthesis, crucial for bacterial survival against this essential but toxic metal.
Area of Science:
- Microbiology
- Molecular Biology
- Biochemistry
Background:
- * Streptococcus pneumoniae is a significant Gram-positive bacterial pathogen responsible for diseases like meningitis.
- * Copper is essential for bacterial growth but toxic at high concentrations.
- * Copper resistance mechanisms are critical virulence factors in microbial pathogens.
Purpose of the Study:
- * To determine the minimum inhibitory concentration (MIC) of copper for Streptococcus pneumoniae growth.
- * To identify proteins involved in copper resistance using proteomic and transcriptomic approaches.
- * To elucidate the role of cellular processes, particularly cell wall biosynthesis, in copper resistance.
Main Methods:
- * Determination of the minimum inhibitory concentration (MIC) of copper.
- * Two-dimensional electrophoresis coupled with mass spectrometry (2D-PAGE/MS) for protein identification.
- * Quantitative reverse transcription PCR (qRT-PCR) for validating proteomic findings.
- * Bioinformatics analysis to categorize differentially expressed proteins.
Main Results:
- * Forty-four proteins showed significant expression changes (p < 0.05, >1.5-fold) in response to copper exposure.
- * Upregulated proteins were primarily involved in cell wall biosynthesis, protein biosynthesis, purine and pyrimidine biosynthesis, and metabolic processes.
- * Quantitative reverse transcription PCR confirmed proteomic data.
Conclusions:
- * The cell wall biosynthesis pathway is a critical determinant of copper resistance in Streptococcus pneumoniae.
- * Copper resistance involves complex regulatory networks affecting multiple cellular processes.
- * Understanding these mechanisms can inform strategies against S. pneumoniae infections.
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