Combining two known antimicrobial peptides (AMPs) into a hybrid peptide is one promising avenue in the design of agents with increased antibacterial activity. However， very few previous studies have considered the effect of creating a hybrid from one AMP that permeabilizes membranes and another AMP that acts intracellularly after translocating across the membrane. Moreover， very few studies have systematically evaluated the order of parent peptides or the presence of linkers in the design of hybrid AMPs. Here， we use a combination of antibacterial measurements， cellular assays and semi-quantitative confocal microscopy to characterize the activity and mechanism for a library of sixteen hybrid peptides. T... More
Combining two known antimicrobial peptides (AMPs) into a hybrid peptide is one promising avenue in the design of agents with increased antibacterial activity. However， very few previous studies have considered the effect of creating a hybrid from one AMP that permeabilizes membranes and another AMP that acts intracellularly after translocating across the membrane. Moreover， very few studies have systematically evaluated the order of parent peptides or the presence of linkers in the design of hybrid AMPs. Here， we use a combination of antibacterial measurements， cellular assays and semi-quantitative confocal microscopy to characterize the activity and mechanism for a library of sixteen hybrid peptides. These hybrids consist of permutations of two primarily membrane translocating peptides， buforin II and DesHDAP1， and two primarily membrane permeabilizing peptides， magainin 2 and parasin. For all hybrids， the permeabilizing peptide appeared to dominate the mechanism， with hybrids primarily killing bacteria through membrane permeabilization. We also observed increased hybrid activity when the permeabilizing parent peptide was placed at the N-terminus. Activity data also highlighted the potential value of considering AMP cocktails in addition to hybrid peptides. Together， these observations will guide future design efforts aiming to design more active hybrid AMPs.
