9C0C

E.coli GroEL apoenzyme

Summary for 9C0C

• Entry DOI: 10.2210/pdb9c0c/pdb
• EMDB information: 45079
• Descriptor: 60 kDa chaperonin (1 entity in total)
• Functional Keywords: groel, chaperone
• Biological source: Escherichia coli
• Total number of polymer chains: 14
• Total formula weight: 803483.95

Authors

Watson, E.R.,Lander, G.C. (deposition date: 2024-05-25, release date: 2024-08-07, Last modification date: 2024-08-14)

Primary citation

Godek, J.,Sivinski, J.,Watson, E.R.,Lebario, F.,Xu, W.,Stevens, M.,Zerio, C.J.,Ambrose, A.J.,Zhu, X.,Trindl, C.A.,Zhang, D.D.,Johnson, S.M.,Lander, G.C.,Chapman, E.
Bis-sulfonamido-2-phenylbenzoxazoles Validate the GroES/EL Chaperone System as a Viable Antibiotic Target.
J.Am.Chem.Soc., 146:20845-20856, 2024

PubMed Abstract: We recently reported on small-molecule inhibitors of the GroES/GroEL chaperone system as potential antibiotics against and the ESKAPE pathogens but were unable to establish GroES/GroEL as the cellular target, leading to cell death. In this study, using two of our most potent -sulfonamido-2-phenylbenzoxazoles (PBZs), we established the binding site of the PBZ molecules using cryo-EM and found that GroEL was the cellular target responsible for the mode of action. Cryo-EM revealed that PBZ1587 binds at the GroEL ring-ring interface (RRI). A cellular reporter assay confirmed that PBZ1587 engaged GroEL in cells, but cellular rescue experiments showed potential off-target effects. This prompted us to explore a closely related analogue, PBZ1038, which is also bound to the RRI. Biochemical characterization showed potent inhibition of Gram-negative chaperonins but much lower potency of chaperonin from a Gram-positive organism, . A cellular reporter assay showed that PBZ1038 also engaged GroEL in cells and that the cytotoxic phenotype could be rescued by a chromosomal copy of GroEL/GroES or by expressing a recalcitrant RRI mutant. These data argue that PBZ1038's antimicrobial action is exerted through inhibition of GroES/GroEL, validating this chaperone system as an antibiotic target.

PubMed: 39041457
DOI: 10.1021/jacs.4c05057

Experimental method

ELECTRON MICROSCOPY (3.41 Å)