Comparative Proteomic Analysis of Hydrogen Peroxide-Induced Protein Expression in Streptococcus pneumoniae D39

Title:Comparative Proteomic Analysis of Hydrogen Peroxide-Induced Protein Expression in Streptococcus pneumoniae D39

Volume: 18 Issue: 5

Author(s): Sungkyoung Lee, Myoung-Ro Lee, Songmee Bae*Min-Kyu Kwak*

Affiliation:

• Division of Antimicrobial Resistance, Research Center for Infectious Disease Research, Korea National Institute of Health, Korea Disease Control and Prevention Agency, Cheongju-si, Chungcheongbuk-do, 28160,Korea

• Laboratory of Microbial Physiology and Biotechnology, Department of Food and Nutrition, Institute of Food and Nutrition Science, Eulji University, Gyeonggi-do, 13135,Korea

Keywords: High-throughput image analysis, hydrogen peroxide, LC-coupled ESI-MS/MS, MALDI-TOF, oxidative stress, Streptococcus pneumoniae, two-dimensional gel electrophoresis.

Abstract:

Background: Streptococcus pneumoniaeis a leading cause of human respiratory tract infection. Despite the lack of activities of antioxidative enzymes, including cytochromes, hemoproteins, and peroxidases/catalases, traits conferring the aerotolerant-anaerobic growth of this bacterium are conserved, with the high efficacy of antioxidative actions, in an oxygen-rich environment.

Objective: Through proteome analysis, this study's intention was to evaluate differentially expressed proteins and/or gene products modeled in a highly virulent strain, S. pneumoniae D39, exogenously- treated with millimolar concentrations of H₂O₂.

Methods: For two-dimensional gel electrophoresis (2-DE) analysis, following one dimensional isoelectric focusing with an immobilized pH gradient of pH 4-7, the most significantly mobilized proteins expressed were separated by SDS-PAGE in the second dimension. With a total of 431 protein spots detected, certain proteins were excised, in-gel trypsin digested, and analyzed by combination with MALDI-TOF and LC-ESI-MS/MS for mass spectrometric peptide mapping and protein identification. Utilizing mass spectrometry analysis of spots excised from 2-DE, the selected protein spots were identified with a variety of databases and MASCOT.

Results: With the aid of comparisons to proteome reference maps, the most differentially expressed 38 proteins, those with approximately 1.4-fold or more increase and/or decrease or with multiple isoforms exhibiting variable pI values, were induced by treatment of exogenous 2 mM H₂O₂.The identified proteins were seen to be involved in pneumococcal pathogenesis and primary metabolism, amongst others.

Conclusion: This is the first study to convincingly document proteomic information associated with pathophysiological adaptation under the given oxidative conditions, and corresponding potential antioxidative mechanisms, in S. pneumoniae.

Cite this article as:

Lee Sungkyoung , Lee Myoung-Ro , Bae Songmee *, Kwak Min-Kyu *, Comparative Proteomic Analysis of Hydrogen Peroxide-Induced Protein Expression in Streptococcus pneumoniae D39, Current Proteomics 2021; 18 (5) . https://dx.doi.org/10.2174/1570164618999210104223526