title: Identification by High-Throughput Screening of Pseudomonas Acyl-Coenzyme A Synthetase Inhibitors.
abstract: Pseudomonas infections are common among hospitalized, immunocompromised, and chronic lung disease patients. These infections are recalcitrant to common antibacterial therapies due to inherent antibiotic resistance. To meet the need of new anti- Pseudomonas drugs, a sensitive, homogenous, and robust assay was developed with the aim of identifying inhibitors of acyl-coenzyme A synthetases (ACSs) from Pseudomonas. Given the importance of fatty acids for in vivo nutrition of Pseudomonas, such inhibitors might have the potential to reduce the bacterial fitness during infection. The assay, based on a coupled reaction between the Pseudomonas spp. ACS and the firefly luciferase, allowed the identification of three classes of inhibitors by screening of a diverse compound collection. These compounds were confirmed to reversibly bind ACS with potencies in the micromolar range. Two classes were found to compete with acyl-coenzyme A, while the third one was competitive with fatty acid binding. Although these compounds inhibit the bacterial ACS in cell-free assays, they show modest or no effect on Pseudomonas growth in vitro.
authors.affiliation: 1 IRBM Science Park S.p.A, Pomezia, Italy.
authors.forename: Lorenzo
authors.initials: L
authors.lastname: Turcano
authors.affiliation: 2 Department of Science, RomaTre University, Rome, Italy.
authors.forename: Daniela
authors.initials: D
authors.lastname: Visaggio
authors.affiliation: 2 Department of Science, RomaTre University, Rome, Italy.
authors.forename: Emanuela
authors.initials: E
authors.lastname: Frangipani
authors.affiliation: 1 IRBM Science Park S.p.A, Pomezia, Italy.
authors.forename: Antonino
authors.initials: A
authors.lastname: Missineo
authors.affiliation: 1 IRBM Science Park S.p.A, Pomezia, Italy.
authors.forename: Matteo
authors.initials: M
authors.lastname: Andreini
authors.affiliation: 1 IRBM Science Park S.p.A, Pomezia, Italy.
authors.forename: Sergio
authors.initials: S
authors.lastname: Altamura
authors.affiliation: 2 Department of Science, RomaTre University, Rome, Italy.
authors.forename: Paolo
authors.initials: P
authors.lastname: Visca
authors.affiliation: 1 IRBM Science Park S.p.A, Pomezia, Italy.
authors.forename: Alberto
authors.initials: A
authors.lastname: Bresciani
mesh_terms: D000214:Acyl Coenzyme A / Q000037:antagonists & inhibitors* / Q000378:metabolism*; D003066:Coenzyme A Ligases / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D005227:Fatty Acids / Q000378:metabolism; D057166:High-Throughput Screening Assays / Q000379:methods; D049409:Luciferases, Firefly / Q000378:metabolism; D011549:Pseudomonas / Q000378:metabolism*; D011552:Pseudomonas Infections / Q000382:microbiology
chemical_list: D000214:Acyl Coenzyme A; D004791:Enzyme Inhibitors; D005227:Fatty Acids; D049409:Luciferases, Firefly; D003066:Coenzyme A Ligases
keywords: Pseudomonas; acyl-CoA synthetase; antibacterial; high-throughput screening; hit identification