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Burkholderia pseudomallei d ‐alanine‐ d ‐alanine ligase; detailed characterisation and assessment of a potential antibiotic drug target

DOI: 10.1111/febs.14976 DOI Help

Authors: Laura Diaz-Saez (University of Dundee) , Leah S. Torrie (University of Dundee) , Stuart P. Mcelroy (University of Dundee) , David Gray (University of Dundee) , William N. Hunter (University of Dundee)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: The Febs Journal , VOL 55

State: Published (Approved)
Published: July 2019
Diamond Proposal Number(s): 10071

Open Access Open Access

Abstract: Burkholderia pseudomallei is a serious, difficult to treat Gram‐negative pathogen and an increase in the occurrence of drug‐resistant strains has been detected. We have directed efforts to identify and to evaluate potential drug targets relevant to treatment of infection by B. pseudomallei. We have selected and characterised the essential enzyme d‐alanine‐d‐alanine ligase (BpDdl), required for the ATP‐assisted biosynthesis of a peptidoglycan precursor. A recombinant supply of protein supported high‐resolution crystallographic and biophysical studies with ligands (AMP and AMP+d‐Ala‐d‐Ala), and comparisons with orthologues enzymes suggest a ligand‐induced conformational change occurring that might be relevant to the catalytic cycle. The detailed biochemical characterisation of the enzyme, development and optimisation of ligand binding assays supported the search for novel inhibitors by screening of selected compound libraries. In a similar manner to that observed previously in other studies, we note a paucity of hits that are worth follow‐up and then in combination with a computational analysis of the active site, we conclude that this ligase represents a difficult target for drug discovery. Nevertheless, our reagents, protocols and data can underpin future efforts exploiting more diverse chemical libraries and structure‐based approaches.

Journal Keywords: antibacterial; enzyme assay; enzyme inhibition; peptidoglycan; target validation

Diamond Keywords: Bacteria; Enzymes

Subject Areas: Biology and Bio-materials, Medicine, Chemistry

Instruments: I04-1-Macromolecular Crystallography (fixed wavelength)

Other Facilities: ESRF

Added On: 24/07/2019 11:49


Discipline Tags:

Pathogens Antibiotic Resistance Infectious Diseases Health & Wellbeing Biochemistry Chemistry Structural biology Organic Chemistry Drug Discovery Life Sciences & Biotech

Technical Tags:

Diffraction Macromolecular Crystallography (MX)