Inhibition of Diverse DsbA Enzymes in Multi-DsbA Encoding Pathogens

Makrina Totsika, Dimitrios Vagenas, Jason J. Paxman, Geqing Wang, Rabeb Dhouib, Pooja Sharma, Jennifer L. Martin, Martin J. Scanlon, Begoña Heras

Research output: Contribution to journalArticleResearchpeer-review

Abstract

Aims: DsbA catalyzes disulfide bond formation in secreted and outer membrane proteins in bacteria. In pathogens, DsbA is a major facilitator of virulence constituting a target for antivirulence antimicrobial development. However, many pathogens encode multiple and diverse DsbA enzymes for virulence factor folding during infection. The aim of this study was to determine whether our recently identified inhibitors of Escherichia coli K-12 DsbA can inhibit the diverse DsbA enzymes found in two important human pathogens and attenuate their virulence. 

Results: DsbA inhibitors from two chemical classes (phenylthiophene and phenoxyphenyl derivatives) inhibited the virulence of uropathogenic E. coli and Salmonella enterica serovar Typhimurium, encoding two and three diverse DsbA homologues, respectively. Inhibitors blocked the virulence of dsbA null mutants complemented with structurally diverse DsbL and SrgA, suggesting that they were not selective for prototypical DsbA. Structural characterization of DsbA-inhibitor complexes showed that compounds from each class bind in a similar region of the hydrophobic groove adjacent to the Cys30-Pro31-His32-Cys33 (CPHC) active site. Modeling of DsbL- and SrgA-inhibitor interactions showed that these accessory enzymes could accommodate the inhibitors in their different hydrophobic grooves, supporting our in vivo findings. Further, we identified highly conserved residues surrounding the active site for 20 diverse bacterial DsbA enzymes, which could be exploited in developing inhibitors with a broad spectrum of activity. 

Innovation and Conclusion: We have developed tools to analyze the specificity of DsbA inhibitors in bacterial pathogens encoding multiple DsbA enzymes. This work demonstrates that DsbA inhibitors can be developed to target diverse homologues found in bacteria. Antioxid. Redox Signal. 29, 653-666.

Original languageEnglish
Pages (from-to)653-666
Number of pages14
JournalAntioxidants and Redox Signaling
Volume29
Issue number7
DOIs
Publication statusPublished - 1 Sep 2018

Keywords

  • Infection
  • protein folding
  • therapeutics
  • thiol

Cite this

Totsika, M., Vagenas, D., Paxman, J. J., Wang, G., Dhouib, R., Sharma, P., ... Heras, B. (2018). Inhibition of Diverse DsbA Enzymes in Multi-DsbA Encoding Pathogens. Antioxidants and Redox Signaling, 29(7), 653-666. https://doi.org/10.1089/ars.2017.7104
Totsika, Makrina ; Vagenas, Dimitrios ; Paxman, Jason J. ; Wang, Geqing ; Dhouib, Rabeb ; Sharma, Pooja ; Martin, Jennifer L. ; Scanlon, Martin J. ; Heras, Begoña. / Inhibition of Diverse DsbA Enzymes in Multi-DsbA Encoding Pathogens. In: Antioxidants and Redox Signaling. 2018 ; Vol. 29, No. 7. pp. 653-666.
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Totsika, M, Vagenas, D, Paxman, JJ, Wang, G, Dhouib, R, Sharma, P, Martin, JL, Scanlon, MJ & Heras, B 2018, 'Inhibition of Diverse DsbA Enzymes in Multi-DsbA Encoding Pathogens' Antioxidants and Redox Signaling, vol. 29, no. 7, pp. 653-666. https://doi.org/10.1089/ars.2017.7104

Inhibition of Diverse DsbA Enzymes in Multi-DsbA Encoding Pathogens. / Totsika, Makrina; Vagenas, Dimitrios; Paxman, Jason J.; Wang, Geqing; Dhouib, Rabeb; Sharma, Pooja; Martin, Jennifer L.; Scanlon, Martin J.; Heras, Begoña.

In: Antioxidants and Redox Signaling, Vol. 29, No. 7, 01.09.2018, p. 653-666.

Research output: Contribution to journalArticleResearchpeer-review

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AB - Aims: DsbA catalyzes disulfide bond formation in secreted and outer membrane proteins in bacteria. In pathogens, DsbA is a major facilitator of virulence constituting a target for antivirulence antimicrobial development. However, many pathogens encode multiple and diverse DsbA enzymes for virulence factor folding during infection. The aim of this study was to determine whether our recently identified inhibitors of Escherichia coli K-12 DsbA can inhibit the diverse DsbA enzymes found in two important human pathogens and attenuate their virulence. Results: DsbA inhibitors from two chemical classes (phenylthiophene and phenoxyphenyl derivatives) inhibited the virulence of uropathogenic E. coli and Salmonella enterica serovar Typhimurium, encoding two and three diverse DsbA homologues, respectively. Inhibitors blocked the virulence of dsbA null mutants complemented with structurally diverse DsbL and SrgA, suggesting that they were not selective for prototypical DsbA. Structural characterization of DsbA-inhibitor complexes showed that compounds from each class bind in a similar region of the hydrophobic groove adjacent to the Cys30-Pro31-His32-Cys33 (CPHC) active site. Modeling of DsbL- and SrgA-inhibitor interactions showed that these accessory enzymes could accommodate the inhibitors in their different hydrophobic grooves, supporting our in vivo findings. Further, we identified highly conserved residues surrounding the active site for 20 diverse bacterial DsbA enzymes, which could be exploited in developing inhibitors with a broad spectrum of activity. Innovation and Conclusion: We have developed tools to analyze the specificity of DsbA inhibitors in bacterial pathogens encoding multiple DsbA enzymes. This work demonstrates that DsbA inhibitors can be developed to target diverse homologues found in bacteria. Antioxid. Redox Signal. 29, 653-666.

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Totsika M, Vagenas D, Paxman JJ, Wang G, Dhouib R, Sharma P et al. Inhibition of Diverse DsbA Enzymes in Multi-DsbA Encoding Pathogens. Antioxidants and Redox Signaling. 2018 Sep 1;29(7):653-666. https://doi.org/10.1089/ars.2017.7104