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Peptide-Based Covalent Inhibitors Bearing Mild Electrophiles to Target a Conserved His Residue of the Bacterial Sliding Clamp

ABSTRACT: Bacterial infections caused by drug-resistant pathogens represent a major threat against which effective antibiotics acting via new mechanisms and engaging new biological targets are highly needed. The sliding clamp (SC), an essential component of the bacterial DNA replication machinery, has been identified as a new target. This processivity factor operates through multiple protein-protein interactions with conserved peptide motifs in multiple proteins involved in DNA replication and repair. Herein, we report the structure-guided design of targeted covalent inhibitors (TCIs) able to bind covalently and selectively to the E. coli SC (EcSC). The inhibitors target His175, a well-conserved SC residue located on the edge of the peptide-binding pocket. Thermodynamic and kinetic analyses confirmed the higher efficiency of the chloroacetamide warhead compared to Michael acceptors. High-resolution X-ray structures of two different covalent inhibitor–SC adducts revealed the canonical ligand orientation and confirmed the formation of the covalent bond with His175. Proteomic studies were consistent with a selective EcSC engagement by the chloroacetamide-based TCI. Finally, the TCI of SC was substantially more active than the parent non-covalent inhibitor in an in vitro SC-dependent replicative assay, suggesting the potential of the approach to target the SC of priority pathogens.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Escherichia Coli

SUBMITTER: Johana Chicher

LAB HEAD: Jérôme Wagner

PROVIDER: PXD042384 | Pride | 2024-01-29

REPOSITORIES: Pride

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			Action	DRS
	2021_S51_JWagner_32-1.raw	Raw
	2021_S51_JWagner_32-2.raw	Raw
	2021_S51_JWagner_32-3.raw	Raw
	2021_S51_JWagner_34-1.raw	Raw
	2021_S51_JWagner_34-2.raw	Raw

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