RT Journal Article SR Electronic T1 A competitive activity-based protein profiling platform yields cell wall synthesis inhibitors active against replicating and non-replicating Mycobacterium tuberculosis JF bioRxiv FD Cold Spring Harbor Laboratory SP 2021.04.16.440156 DO 10.1101/2021.04.16.440156 A1 Michael Li A1 Hiren V. Patel A1 Armand B. Cognetta III A1 Trever C. Smith II A1 Ivy Mallick A1 Jean-François Cavalier A1 Stephane Canaan A1 Bree B. Aldridge A1 Benjamin F. Cravatt A1 Jessica C. Seeliger YR 2021 UL http://biorxiv.org/content/early/2021/04/16/2021.04.16.440156.abstract AB The identification and validation of a small molecule’s targets is a major bottleneck in the discovery process for tuberculosis antibiotics. Activity-based protein profiling (ABPP) is an efficient tool for determining a small molecule’s targets within complex proteomes. However, how target inhibition relates to biological activity is often left unexplored. Here we studied the effects of 1,2,3-triazole ureas on Mycobacterium tuberculosis (Mtb). After screening ~200 compounds, we focused on two inhibitors active against both exponentially replicating and hypoxia-induced drug-tolerant Mtb that form part of a four-compound structure-activity series. The compound with negligible activity revealed potential false positive targets not addressed in other ABPP studies. Biochemistry, computational docking, and morphological analysis confirmed that active compounds preferentially inhibit serine hydrolases with cell wall and lipid metabolism functions and that disruption of the cell wall underlies biological activity. Our findings showed that ABPP identifies the targets most likely relevant to a compound’s antibacterial activity.Competing Interest StatementThe authors have declared no competing interest.