@article{e1ba71a9e4014b15878a1f2fc02d91a3,
title = "Activity-Based Protein Profiling Reveals That Cephalosporins Selectively Active on Non-replicating Mycobacterium tuberculosis Bind Multiple Protein Families and Spare Peptidoglycan Transpeptidases",
abstract = "As β-lactams are reconsidered for the treatment of tuberculosis (TB), their targets are assumed to be peptidoglycan transpeptidases, as verified by adduct formation and kinetic inhibition of Mycobacterium tuberculosis (Mtb) transpeptidases by carbapenems active against replicating Mtb. Here, we investigated the targets of recently described cephalosporins that are selectively active against non-replicating (NR) Mtb. NR-active cephalosporins failed to inhibit recombinant Mtb transpeptidases. Accordingly, we used alkyne analogs of NR-active cephalosporins to pull down potential targets through unbiased activity-based protein profiling and identified over 30 protein binders. None was a transpeptidase. Several of the target candidates are plausibly related to Mtb{\textquoteright}s survival in an NR state. However, biochemical tests and studies of loss of function mutants did not identify a unique target that accounts for the bactericidal activity of these beta-lactams against NR Mtb. Instead, NR-active cephalosporins appear to kill Mtb by collective action on multiple targets. These results highlight the ability of these β-lactams to target diverse classes of proteins.",
keywords = "ABPP, M. tuberculosis, cephalosporin, click chemistry, non-replicating, β-lactams",
author = "{Lopez Quezada}, Landys and Robert Smith and Lupoli, {Tania J.} and Zainab Edoo and Xiaojun Li and Ben Gold and Julia Roberts and Yan Ling and Park, {Sae Woong} and Quyen Nguyen and Schoenen, {Frank J.} and Kelin Li and Hugonnet, {Jean Emmanuel} and Michel Arthur and Sacchettini, {James C.} and Carl Nathan and Jeffrey Aub{\'e}",
note = "Funding Information: We thank Kristin Burns-Huang (Weill Cornell Medicine) for advice and editorial comments. We are indebted to Sabine Ehrt and Weizhen Xu (Weill Cornell Medicine) for the ΔfecB mutant and complemented strain, Milica Tesic Mark and Henrik Molina (Proteomics Resource Center, Rockefeller University) for assistance, Carolina Adura from the Rockefeller University High Throughput Screening Core for her guidance on DSF, Jamie Bean for his assistance in with sequencing the ΔhtpG Mtb strain, Scott Gradia (University of California, Berkeley) for his gift of pETHis6 Sumo TIV LIC cloning vector, and Deborah Hung (Broad Institute) for generously sharing a transposon mutant library of Mtb. Creation of the transposon library was supported by the Broad Institute Tuberculosis donor group and the Pershing Square Foundation. Funding. This work was supported by the NIH Tri-Institutional TB Research Unit grant U19AI111143 and the Milstein Program in Chemical Biology and Translational Medicine. The Department of Microbiology and Immunology is supported by the William Randolph Hearst Trust. Work from the Department of Biochemistry and Biophysics at Texas Agricultural and Mechanical University was supported by the generosity of the Welch Foundation (A-0015). Publisher Copyright: {\textcopyright} Copyright {\textcopyright} 2020 Lopez Quezada, Smith, Lupoli, Edoo, Li, Gold, Roberts, Ling, Park, Nguyen, Schoenen, Li, Hugonnet, Arthur, Sacchettini, Nathan and Aub{\'e}.",
year = "2020",
month = jun,
day = "23",
doi = "10.3389/fmicb.2020.01248",
language = "English (US)",
volume = "11",
journal = "Frontiers in Microbiology",
issn = "1664-302X",
publisher = "Frontiers Media S. A.",
}