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Category: Data

Description: Here, we set up high-throughput multiprotein assays to screen a large collection (>25K) of structurally diverse small molecules against the Mtb DnaK chaperone system including a J-protein and GrpE. We measure the level of ATP hydrolysis by this system as a proxy to detect small molecule-mediated modulation. Like others, we aimed to identify allosteric inhibitors that might affect DnaK function through distal site or cofactor binding, instead of ATP competitive inhibitors that might exhibit promiscuity against the plethora of cellular nucleotide binding proteins. In doing so, we identify several small molecules that affect the activity of the Mtb DnaK chaperone complex, and focus on two unique compounds that have been used as clinical drugs. Both of these molecules represent mimics of substrates that bind to the NBD or SBD of DnaK, but they lead to proposed conformational changes that inactive the protein. We show that one of these molecules has activity against mycobacterial cells that have compromised folding function. Our work contributes to a small but growing list of chemical scaffolds that have activity against protein chaperones, and strengthens our understanding of how to achieve specificity in bacterial DnaK targeting.