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Article

Development of a High Throughput Screen for the Identification of Inhibitors of Peptidoglycan O-Acetyltransferases, New Potential Antibacterial Targets

Department of Molecular and Cellular Biology, University of Guelph, Guelph, ON N1G 2W1, Canada
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Author to whom correspondence should be addressed.
Antibiotics 2019, 8(2), 65; https://doi.org/10.3390/antibiotics8020065
Received: 1 May 2019 / Revised: 22 May 2019 / Accepted: 22 May 2019 / Published: 27 May 2019
(This article belongs to the Special Issue Novel Strategies against Pathogenic Bacteria)
The O-acetylation of peptidoglycan occurs in many Gram-negative and most Gram-positive pathogens and this modification to the essential wall polymer controls the lytic activity of the autolysins, particularly the lytic transglycosylases, and inhibits that of the lysozymes of innate immunity systems. As such, the peptidoglycan O-acetyltransferases PatA/B and OatA are recognized as virulence factors. In this study, we present the high throughput screening of small compound libraries to identify the first known inhibitors of these enzymes. The fluorometric screening assay developed involved monitoring the respective O-acetyltransferases as esterases using 4-methylumbelliferylacetate as substrate. Pilot screens of 3921 compounds validated the usefulness of the HTS protocol. A number of potential inhibitors were identified amongst a total of 145,000 low molecular-weight compounds, some of which were common to both enzymes, while others were unique to each. After eliminating a number of false positives in secondary screens, dose response curves confirmed the apparent specificity of a benzothiazolyl-pyrazolo-pyridine as an inhibitor of Neisseria gonorrhoeae PatB, and several coumarin-based compounds as inhibitors of both this PatB and OatA from Staphylococcus aureus. The benzothiazolyl-pyrazolo-pyridine was determined to be a non-competitive inhibitor of PatB with a Ki of 126 µM. At 177 µg/mL and close to its solubility limit, this compound caused a 90% reduction in growth of N. gonorrhoeae, while growth of Escherichia coli, a bacterium that lacks PatB and, hence, does not produce O-acetylated peptidoglycan, was unaffected. These data provide preliminary proof of concept that peptidoglycan O-acetyltransferases would serve as useful antibacterial targets. View Full-Text
Keywords: peptidoglycan; O-acetyltransferase; antibiotic target; high throughput screening; HTS peptidoglycan; O-acetyltransferase; antibiotic target; high throughput screening; HTS
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MDPI and ACS Style

Brott, A.S.; Jones, C.S.; Clarke, A.J. Development of a High Throughput Screen for the Identification of Inhibitors of Peptidoglycan O-Acetyltransferases, New Potential Antibacterial Targets. Antibiotics 2019, 8, 65. https://doi.org/10.3390/antibiotics8020065

AMA Style

Brott AS, Jones CS, Clarke AJ. Development of a High Throughput Screen for the Identification of Inhibitors of Peptidoglycan O-Acetyltransferases, New Potential Antibacterial Targets. Antibiotics. 2019; 8(2):65. https://doi.org/10.3390/antibiotics8020065

Chicago/Turabian Style

Brott, Ashley S., Carys S. Jones, and Anthony J. Clarke 2019. "Development of a High Throughput Screen for the Identification of Inhibitors of Peptidoglycan O-Acetyltransferases, New Potential Antibacterial Targets" Antibiotics 8, no. 2: 65. https://doi.org/10.3390/antibiotics8020065

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