Title Resurrecting Inactive Antimicrobial Peptides from the Lipopolysaccharide Trap Resurrecting Inactive Antimicrobial Peptides from Lipopolysaccharide (lps) Trap Singapore 637551, Singapore

26 Host defense antimicrobial peptides (AMPs) are a promising source of antibiotics for the 27 treatment of multiple drug-resistant pathogens. Lipopolysaccharide (LPS), the major 28 component of the outer leaflet of the outer membrane of Gram-negative bacteria, functions as 29 a permeability barrier against a variety of molecules including antimicrobial peptides (AMPs). 30 Further, LPS or endotoxin is the causative agent of sepsis killing 100,000 people per year in 31 the USA alone. LPS can restrict activity of AMPs inducing aggregations at the outer 32 membrane as observed for frog AMPs temporins and also in model AMPs. Aggregated 33 AMPs, as ‘trapped’ by the outer membrane, are unable to traverse through the cell wall 34 causing their inactivation. In this work, we show that these inactive AMPs can overcome LPS 35 induced aggregations while conjugated with a short LPS binding β-boomerang peptide motif 36 and become highly bactericidal. The generated hybrid peptides exhibit activity against Gram37 negative and Gram-positive bacteria in high-salt and detoxify endotoxin. Structural and 38 biophysical studies establish mechanism of action of these peptides in LPS outer membranes. 39 Most importantly, this study provides a new concept for the development of potent broad 40 spectrum antibiotic with efficient outer membrane disruption as the mode of action. 41