Crystal Structure of the Dithiol Oxidase DsbA Enzyme from Proteus Mirabilis Bound Non-covalently to an Active Site Peptide Ligand.

Kurth, Fabian, Duprez, Wilko, Premkumar, Lakshmanane, Schembri, Mark A., Fairlie, David P. and Martin, Jennifer L. (2014) Crystal Structure of the Dithiol Oxidase DsbA Enzyme from Proteus Mirabilis Bound Non-covalently to an Active Site Peptide Ligand.. Journal of Biological Chemistry, 289 28: 19810-19822. doi:10.1074/jbc.M114.552380

Attached Files (Some files may be inaccessible until you login with your UQ eSpace credentials)
Name Description MIMEType Size Downloads
UQ330262_OA.pdf Full text (open access) application/pdf 2.25MB 0

Author Kurth, Fabian
Duprez, Wilko
Premkumar, Lakshmanane
Schembri, Mark A.
Fairlie, David P.
Martin, Jennifer L.
Title Crystal Structure of the Dithiol Oxidase DsbA Enzyme from Proteus Mirabilis Bound Non-covalently to an Active Site Peptide Ligand.
Formatted title
Crystal Structure of the Dithiol Oxidase DsbA Enzyme From Proteus Mirabilis Bound Non-Covalently to an Active Site Peptide Ligand 
Journal name Journal of Biological Chemistry   Check publisher's open access policy
ISSN 0021-9258
Publication date 2014-05-15
Year available 2014
Sub-type Article (original research)
DOI 10.1074/jbc.M114.552380
Open Access Status File (Publisher version)
Volume 289
Issue 28
Start page 19810
End page 19822
Total pages 13
Place of publication Bethesda, MD, United States
Publisher American Society for Biochemistry and Molecular Biology
Language eng
Abstract The disulfide bond forming DsbA enzymes and their DsbB interaction partners are attractive targets for development of antivirulence drugs because both are essential for virulence factor assembly in Gram-negative pathogens. Here we characterize PmDsbA from Proteus mirabilis, a bacterial pathogen increasingly associated with multidrug resistance. PmDsbA exhibits the characteristic properties of a DsbA, including an oxidizing potential, destabilizing disulfide, acidic active site cysteine, and dithiol oxidase catalytic activity. We evaluated a peptide, PWATCDS, derived from the partner protein DsbB and showed by thermal shift and isothermal titration calorimetry that it binds to PmDsbA. The crystal structures of PmDsbA, and the active site variant PmDsbAC30S were determined to high resolution. Analysis of these structures allows categorization of PmDsbA into the DsbA class exemplified by the archetypal Escherichia coli DsbA enzyme. We also present a crystal structure of PmDsbAC30S in complex with the peptide PWATCDS. The structure shows that the peptide binds non-covalently to the active site CXXC motif, the cis-Pro loop, and the hydrophobic groove adjacent to the active site of the enzyme. This high-resolution structural data provides a critical advance for future structure-based design of non-covalent peptidomimetic inhibitors. Such inhibitors would represent an entirely new antibacterial class that work by switching off the DSB virulence assembly machinery.
Formatted abstract
Background: DsbA enzymes assemble bacterial virulence factors and are targets for an entirely new drug class.

Results: Proteus mirabilis DsbA was characterized and its structure determined with a peptide bound non-covalently at the active site.

Conclusion: The structure provides the all-important basis for future inhibitor design.

Significance: New drugs to treat superbugs are urgently needed. DsbA inhibitors could have antivirulence activity against bacterial pathogens.
Keyword Oxidative folding
Thioredoxin fold
Dithiol oxidase
Virulence
Crystal structure
Q-Index Code C1
Q-Index Status Confirmed Code
Grant ID FL0992138
FT100100662
1027369
455829
Institutional Status UQ
Additional Notes PMID: 24831013

Document type: Journal Article
Sub-type: Article (original research)
Collections: Official 2015 Collection
School of Chemistry and Molecular Biosciences
Institute for Molecular Bioscience - Publications
 
Versions
Version Filter Type
Citation counts: TR Web of Science Citation Count  Cited 5 times in Thomson Reuters Web of Science Article | Citations
Scopus Citation Count Cited 6 times in Scopus Article | Citations
Google Scholar Search Google Scholar
Created: Sun, 18 May 2014, 19:15:40 EST by Prem Lakshmanane on behalf of Institute for Molecular Bioscience