Isolation and identification of putative protein substrates of the AAA+ molecular chaperone ClpB from the pathogenic spirochaete Leptospira interrogans

Joanna Krajewska , Zbigniew Arent , Michal Zolkiewski , Sabina Kędzierska-Mieszkowska


Bacterial ClpB is an ATP-dependent Hsp100 chaperone that reactivates aggregated proteins in cooperation with the DnaK chaperone system and promotes survival of bacteria under stress conditions. A large number of publications also indicate that ClpB supports the virulence of bacteria, including a pathogenic spirochaete Leptospira interrogans responsible for leptospirosis in both animals and humans. However, the exact role of ClpB in bacterial pathogenicity remains poorly characterized. It can be assumed that ClpB, due to its role as the molecular chaperone, mediates refolding of essential bacterial proteins, including the known virulence factors, which may become prone to aggregation under infection-induced stresses. In this study, we identified putative substrates of ClpB from L. interrogans (ClpBLi). For this purpose, we used a proteomic approach combining the ClpB-Trap affinity pull-down assays, Liquid chromatography-tandem mass spectrometry (LC-MS-MS/MS), and bioinformatics analyses. Most of the identified proteins were enzymes predominantly associated with major metabolic pathways like the tricarboxylic acid (TCA) cycle, glycolysis–gluconeogenesis and amino acid and fatty acid metabolism. Based on our proteomic study, we suggest that ClpB can support the virulence of L. interrogans by protecting the conformational integrity and catalytic activity of multiple metabolic enzymes, thus maintaining energy homeostasis in pathogen cells.
Author Joanna Krajewska (FB / DB)
Joanna Krajewska,,
- Department of Biochemistry
, Zbigniew Arent
Zbigniew Arent,,
, Michal Zolkiewski
Michal Zolkiewski,,
, Sabina Kędzierska-Mieszkowska (FB / DB)
Sabina Kędzierska-Mieszkowska,,
- Department of Biochemistry
Journal seriesInternational Journal of Molecular Sciences, ISSN 1661-6596, e-ISSN 1422-0067, (A 30 pkt)
Issue year2018
Publication size in sheets0.6
Keywords in Englishbacterial pathogens, ClpB, Leptospira interrogans, leptospirosis, molecular chaperones
Languageen angielski
LicenseJournal (articles only); published final; Uznanie Autorstwa (CC-BY); with publication
Score (nominal)30
ScoreMinisterial score = 30.0, ArticleFromJournal
Ministerial score (2013-2016) = 30.0, ArticleFromJournal
Publication indicators WoS Impact Factor: 2017 = 3.687 (2) - 2017=3.878 (5)
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* presented citation count is obtained through Internet information analysis and it is close to the number calculated by the Publish or Perish system.