Back to Search
Start Over
Targeting the Bacterial Epitranscriptome for Antibiotic Development: Discovery of Novel tRNA-(N 1 G37) Methyltransferase (TrmD) Inhibitors.
- Source :
-
ACS infectious diseases [ACS Infect Dis] 2019 Mar 08; Vol. 5 (3), pp. 326-335. Date of Electronic Publication: 2019 Feb 04. - Publication Year :
- 2019
-
Abstract
- Bacterial tRNA modification synthesis pathways are critical to cell survival under stress and thus represent ideal mechanism-based targets for antibiotic development. One such target is the tRNA-(N <superscript>1</superscript> G37) methyltransferase (TrmD), which is conserved and essential in many bacterial pathogens. Here we developed and applied a widely applicable, radioactivity-free, bioluminescence-based high-throughput screen (HTS) against 116350 compounds from structurally diverse small-molecule libraries to identify inhibitors of Pseudomonas aeruginosa TrmD ( PaTrmD). Of 285 compounds passing primary and secondary screens, a total of 61 TrmD inhibitors comprised of more than 12 different chemical scaffolds were identified, all showing submicromolar to low micromolar enzyme inhibitor constants, with binding affinity confirmed by thermal stability and surface plasmon resonance. S-Adenosyl-l-methionine (SAM) competition assays suggested that compounds in the pyridine-pyrazole-piperidine scaffold were substrate SAM-competitive inhibitors. This was confirmed in structural studies, with nuclear magnetic resonance analysis and crystal structures of PaTrmD showing pyridine-pyrazole-piperidine compounds bound in the SAM-binding pocket. Five hits showed cellular activities against Gram-positive bacteria, including mycobacteria, while one compound, a SAM-noncompetitive inhibitor, exhibited broad-spectrum antibacterial activity. The results of this HTS expand the repertoire of TrmD-inhibiting molecular scaffolds that show promise for antibiotic development.
- Subjects :
- Anti-Bacterial Agents chemistry
Bacterial Proteins chemistry
Bacterial Proteins genetics
Bacterial Proteins metabolism
Drug Discovery
Enzyme Inhibitors chemistry
Kinetics
Methyltransferases chemistry
Methyltransferases genetics
Methyltransferases metabolism
Pseudomonas aeruginosa genetics
Substrate Specificity
Anti-Bacterial Agents pharmacology
Bacterial Proteins antagonists & inhibitors
Drug Evaluation, Preclinical methods
Enzyme Inhibitors pharmacology
Methyltransferases antagonists & inhibitors
Pseudomonas aeruginosa drug effects
Pseudomonas aeruginosa enzymology
RNA, Transfer metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 2373-8227
- Volume :
- 5
- Issue :
- 3
- Database :
- MEDLINE
- Journal :
- ACS infectious diseases
- Publication Type :
- Academic Journal
- Accession number :
- 30682246
- Full Text :
- https://doi.org/10.1021/acsinfecdis.8b00275