%0 Journal Article %A Justin R. Randall %A Cory D. DuPai %A T. Jeffrey Cole %A Gillian Davidson %A Kyra E. Groover %A Claus O. Wilke %A Bryan W. Davies %T Designing β-hairpin peptide macrocycles for antibiotic potential %D 2022 %R 10.1101/2022.06.21.497034 %J bioRxiv %P 2022.06.21.497034 %X Peptide macrocycles are a rapidly emerging new class of therapeutic, yet the design of their structure and activity remains challenging. This is especially true for those with β-hairpin structure due to weak folding properties and a propensity for aggregation. Here we use proteomic analysis and common antimicrobial features to design a large peptide library with macrocyclic β-hairpin structure. Using an activity-driven high-throughput screen we identify dozens of peptides killing bacteria through selective membrane disruption and analyze their biochemical features via machine learning. Active peptides contain a unique constrained structure and are highly enriched for cationic charge with arginine in their turn region. Our results provide a synthetic strategy for structured macrocyclic peptide design and discovery, while also elucidating characteristics important for β-hairpin antimicrobial peptide activity.Brief Summary We design, screen, and computationally analyze a synthetic macrocyclic β-hairpin peptide library for antibiotic potential.Competing Interest StatementThe authors have declared no competing interest. %U https://www.biorxiv.org/content/biorxiv/early/2022/06/22/2022.06.21.497034.full.pdf