High-Throughput Identification and Characterization of LptDE-Binding Bicycle Peptides Using Phage Display and Cryo-EM
Year: 2025
Type: article
Abstract: The lipopolysaccharide (LPS) transport (Lpt) system in Gram-negative bacteria maintains the integrity of the asymmetric bacterial outer membrane (OM). LPS biogenesis systems are essential in most Gram-negative bacteria, with LptDE responsible for the delivery of LPS to the outer leaflet of the OM. As an externally accessible, essential protein, LptDE offers a promising target for inhibitor development without the need for cellular penetration. However, there are no direct inhibitors of E. coli LptDE, and drug discovery is made challenging since it is a membrane target without a conventional active site. Here, the bicycle phage display platform was used in combination with cryogenic-electron microscopy (cryo-EM) and surface plasmon resonance to identify and map bicyclic peptide binders to Shigella flexneri LptDE (SfLptDE). Four distinct epitopes with unique bicycle molecule binding motifs were identified across the SfLptD β-barrel. This method represents a streamlined workflow for the identification and prioritization of hit molecules against LptDE.
Source: Journal of Medicinal Chemistry
Authors: Shenaz Allyjaun, Emily Dunbar, Steven W. Hardwick, Sarah Newell, Finn Holding, Catherine E Rowland, Megan A. St. Denis, Simone Pellegrino, Gustavo Arruda Bezerra, Nikolaos Bournakas, Dimitri Y. Chirgadze, Lee Cooper, Giulia Paris, Nick Lewis, Peter Brown, Michael J. Skynner, Michael J Dawson, Paul Beswick, Julia Hubbard, Bert van den Berg, Hector Newman
Institutions: Newcastle University, University of Cambridge
Language: English
Subfield: Genetics
Domain: Life Sciences
Open Access status: hybrid
Estimated APC paid: $4,500
OpenAlex ID: W4414865294