Targeting the Bacterial Epitranscriptome for Antibiotic Development: Discovery of Novel tRNA-(N1G37) Methyltransferase (TrmD) Inhibitors
| العنوان: | Targeting the Bacterial Epitranscriptome for Antibiotic Development: Discovery of Novel tRNA-(N1G37) Methyltransferase (TrmD) Inhibitors |
|---|---|
| المؤلفون: | Yee Hwa Wong, CongBao Kang, Xiaoying Koh-Stenta, Alex Matter, Wenhe Zhong, Hui Qi Ng, Klement Foo, Megan E. McBee, Ann Koay, Anders Poulsen, Julien Lescar, Jeffrey Hill, Abbas El Sahili, Qianhui Nah, Yan Li, Yok Hian Chionh, Anna Ngo, Meng Ling Choong, Peter C. Dedon |
| المساهمون: | School of Biological Sciences, NTU Institute of Structural Biology, Singapore-MIT Alliance for Research and Technology |
| المصدر: | ACS Infectious Diseases. 5:326-335 |
| بيانات النشر: | American Chemical Society (ACS), 2019. |
| سنة النشر: | 2019 |
| مصطلحات موضوعية: | 0301 basic medicine, TRNA modification, Methyltransferase, biology, Chemistry, High-throughput screening, 030106 microbiology, Biological sciences [Science], tRNA Modification, biology.organism_classification, 03 medical and health sciences, 030104 developmental biology, Infectious Diseases, Biochemistry, Enzyme inhibitor, High-throughput Screening, Transfer RNA, biology.protein, Transferase, Antibacterial activity, Bacteria |
| الوصف: | 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-(N1G37) 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. Agency for Science, Technology and Research (A*STAR) Ministry of Education (MOE) National Research Foundation (NRF) Singapore-MIT Alliance for Research and Technology (SMART) Published version This research was supported by the National Research Foundation of Singapore through the Singapore-MIT Alliance for Research and Technology (SMART) Infectious Diseases and Antimicrobial Resistance Interdisciplinary Research Groups; SMART Innovation Centre Grant ING137070-BIO to P.D. and J.L.; the Biomedical Sciences Institutes (BMSI), Agency for Science, Technology, and Research (A*STAR), Singapore; and AcRF Grants Tier1 RG154/14 and MOE2015- T2-2-075 to J.L. |
| وصف الملف: | application/pdf |
| تدمد: | 2373-8227 |
| DOI: | 10.1021/acsinfecdis.8b00275 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::1ba136b2d4dbc1e07c0350f39a81a8c5 https://doi.org/10.1021/acsinfecdis.8b00275 |
| Rights: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....1ba136b2d4dbc1e07c0350f39a81a8c5 |
| قاعدة البيانات: | OpenAIRE |
كن أول من يترك تعليقا!