Sent packing: Protein engineering generates a new crystal form of Pseudomonas aeruginosa DsbA1 with increased catalytic surface accessibility

McMahon, Roisin M., Coincon, Mathieu, Tay, Stephanie, Heras, Begona, Morton, Craig J., Scanlon, Martin J. and Martin, Jennifer L. (2015) Sent packing: Protein engineering generates a new crystal form of Pseudomonas aeruginosa DsbA1 with increased catalytic surface accessibility. Acta Crystallographica Section D: Biological Crystallography, 71 2386-2395. doi:10.1107/S1399004715018519


Author McMahon, Roisin M.
Coincon, Mathieu
Tay, Stephanie
Heras, Begona
Morton, Craig J.
Scanlon, Martin J.
Martin, Jennifer L.
Title Sent packing: Protein engineering generates a new crystal form of Pseudomonas aeruginosa DsbA1 with increased catalytic surface accessibility
Journal name Acta Crystallographica Section D: Biological Crystallography   Check publisher's open access policy
ISSN 1399-0047
Publication date 2015
Year available 2015
Sub-type Article (original research)
DOI 10.1107/S1399004715018519
Open Access Status DOI
Volume 71
Start page 2386
End page 2395
Total pages 10
Place of publication Malden, MA, United States
Publisher Wiley-Blackwell Publishing
Collection year 2016
Language eng
Formatted abstract
Pseudomonas aeruginosa is an opportunistic human pathogen for which new antimicrobial drug options are urgently sought. P. aeruginosa disulfide-bond protein A1 (PaDsbA1) plays a pivotal role in catalyzing the oxidative folding of multiple virulence proteins and as such holds great promise as a drug target. As part of a fragment-based lead discovery approach to PaDsbA1 inhibitor development, the identification of a crystal form of PaDsbA1 that was more suitable for fragment-soaking experiments was sought. A previously identified crystallization condition for this protein was unsuitable, as in this crystal form of PaDsbA1 the active-site surface loops are engaged in the crystal packing, occluding access to the target site. A single residue involved in crystal-packing interactions was substituted with an amino acid commonly found at this position in closely related enzymes, and this variant was successfully used to generate a new crystal form of PaDsbA1 in which the active-site surface is more accessible for soaking experiments. The PaDsbA1 variant displays identical redox character and in vitro activity to wild-type PaDsbA1 and is structurally highly similar. Two crystal structures of the PaDsbA1 variant were determined in complex with small molecules bound to the protein active site. These small molecules (MES, glycerol and ethylene glycol) were derived from the crystallization or cryoprotectant solutions and provide a proof of principle that the reported crystal form will be amenable to co-crystallization and soaking with small molecules designed to target the protein active-site surface.
Keyword Bacterial virulence
Drug design
Fragment-based lead discovery
DsbA
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: Official 2016 Collection
Institute for Molecular Bioscience - Publications
 
Versions
Version Filter Type
Citation counts: TR Web of Science Citation Count  Cited 0 times in Thomson Reuters Web of Science Article
Scopus Citation Count Cited 0 times in Scopus Article
Google Scholar Search Google Scholar
Created: Sun, 27 Dec 2015, 00:26:45 EST by System User on behalf of Scholarly Communication and Digitisation Service