A Cell-based Screen in Actinomyces oris to Identify Sortase Inhibitors

Sortase enzymes are attractive antivirulence drug targets that attach virulence factors to the surface of Staphylococcus aureus and other medically significant bacterial pathogens. Prior efforts to discover a useful sortase inhibitor have relied upon an in vitro activity assay in which the enzyme is...

Full description

Saved in:
Bibliographic Details
Published in:Scientific reports 2020-05, Vol.10 (1), p.8520-8520, Article 8520
Main Authors: Gosschalk, Jason E., Chang, Chungyu, Sue, Christopher K., Siegel, Sara D., Wu, Chenggang, Kattke, Michele D., Yi, Sung Wook, Damoiseaux, Robert, Jung, Michael E., Ton-That, Hung, Clubb, Robert T.
Format: Article
Language:English
Subjects:
631/154/1435/2163
631/326
631/326/22
Actinomyces
Actinomyces - drug effects
Actinomyces - enzymology
Actinomyces - growth & development
Aminoacyltransferases - antagonists & inhibitors
Aminoacyltransferases - metabolism
Bacterial Proteins - antagonists & inhibitors
Bacterial Proteins - metabolism
Biofilms
Biofilms - drug effects
Biofilms - growth & development
Cell culture
Cells, Cultured
Enzymatic activity
Enzyme Inhibitors - pharmacology
Enzymes
High-throughput screening
High-Throughput Screening Assays - methods
Humanities and Social Sciences
Inhibitors
multidisciplinary
Phenotypes
Science
Science (multidisciplinary)
Sortase
Therapeutic targets
Virulence factors
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Sortase enzymes are attractive antivirulence drug targets that attach virulence factors to the surface of Staphylococcus aureus and other medically significant bacterial pathogens. Prior efforts to discover a useful sortase inhibitor have relied upon an in vitro activity assay in which the enzyme is removed from its native site on the bacterial surface and truncated to improve solubility. To discover inhibitors that are effective in inactivating sortases in vivo , we developed and implemented a novel cell-based screen using Actinomyces oris , a key colonizer in the development of oral biofilms. A . oris is unique because it exhibits sortase-dependent growth in cell culture, providing a robust phenotype for high throughput screening (HTS). Three molecules representing two unique scaffolds were discovered by HTS and disrupt surface protein display in intact cells and inhibit enzyme activity in vitro . This represents the first HTS for sortase inhibitors that relies on the simple metric of cellular growth and suggests that A . oris may be a useful platform for discovery efforts targeting sortase.
ISSN:2045-2322
2045-2322
DOI:10.1038/s41598-020-65256-x