Systematic Analysis of Hybrid Antimicrobial Peptides

Abstract

Antimicrobial peptides (AMPs) are part of the immune response of all classes of life and have gained attention as promising alternative treatments for infectious bacteria resistant to conventional antibiotics. AMPs kill bacteria through two known mechanisms of action. Some AMPs, such as parasin and magainin II, kill bacteria by inducing membrane permeabilization. Other AMPs, such as buforin II (BF2) and DesHDAP1, readily translocate across the membrane and interact with intracellular components including nucleic acids. In recent years, there has been increased interest in developing hybrid AMPs that combine two distinct AMPs into a single peptide. These hybrid AMPs have been shown to be more potent than their individual AMP components. To date, few studied hybrid have combined AMPs that follow different mechanisms. Here, we focus on using a variety of cellular assays and confocal imaging to characterize the activity and mechanisms of action of hybrid AMPs that combine one permeabilizing AMP (parasin or magainin II) with one translocating AMP (BF2 or DesHDAP1) in different orientations and with different linkers. We show that these hybrid AMPs are generally more potent than their individual AMP components and that the permeabilizing peptide (parasin or magainin II) dominates the mechanism of action when combined with the translocating peptide (BF2 or DesHDAP1). These observations of 16 hybrid peptides have elucidated trends that will promote the rational design of AMPs with enhanced activity.

Rational Designed Hybrid Peptides Show up to a 6-fold Increase in Antimicrobial Activity and Demonstrate Different Ultrastructural Changes as the Parental Peptides measured by BioSAXS

“…Based on the success of the hybridization strategy reported by others, for example (Fox et al, 2012;Shang et al, 2020;Wade et al, 2018Wade et al, , 2019Xu et al, 2014), we hypothesized that hybrid molecules from two different classes of AMPs, which we have identified in the past as potential antimicrobial drug candidates, may lead to a further improvement of antimicrobial activity. Thus we combined the excellent activity of proline-rich and arginine-isoleucine-rich AMPs.…”

“…Antimicrobial peptides are a very broad and diverse class of compounds that can kill multi-drug resistant bacteria and therefore have the potential to become the next generation of antibiotics. Some hybrid AMPs have been shown to possess improved activity (Shang et al, 2020;Wade et al, 2018Wade et al, , 2019) and here we explored the combination of two different classes of peptides, proline-rich peptides, and arginine-isoleucine-rich peptides. While the mode of action of the prolinerich peptides is known, the mode of action of the arginine-isoleucine-rich peptides has yet to be discovered.…”

“…The peptide optP7 was used as a lead and selected for studying lipidation, glycosylation, cyclisation, and grafting into a cyclotide (Grimsey et al, 2020;Koehbach et al, 2021). Hybridization of different AMPs has been investigated to further improve their performance (Shang et al, 2020;Wade et al, 2018Wade et al, , 2019. Here we report the creation of different hybrid molecules using peptides optP1 (KIILRIRWR) and optP7 (KRRVWIIW) in combination with the Bac5(1-17) variant 291 (RWRRPIRRRPIRPPFWR-CONH2) and a consensus sequence from proline-rich antimicrobial peptides, called consP1 (VRKPPYLPRPRPRPL-CONH2).…”

Rational Designed Hybrid Peptides Show up to a 6-Fold Increase in Antimicrobial Activity and Demonstrate Different Ultrastructural Changes as the Parental Peptides Measured by BioSAXS

“…Antimicrobial peptides are a very broad and diverse class of compounds that can kill multi-drug resistant bacteria and therefore have the potential to become the next generation of antibiotics. Based on the success of the hybridization strategy reported by others, for example (Fox et al, 2012;Xu et al, 2014;Wade et al, 2018Wade et al, , 2019Shang et al, 2020), we hypothesized that hybrid molecules from two different classes of AMPs, which we have identified in the past as potential antimicrobial drug candidates, may lead to a further improvement of antimicrobial activity. Thus we combined the excellent activity of proline-rich and arginine-isoleucine-rich AMPs.…”

Mini Review on Antimicrobial Peptides, Sources, Mechanism and Recent Applications

“…magainin is being used in treating viral and bacterial diseases [ 151 , 152 ]. Several studies carried out globally by researchers have provided in depth understanding about antimicrobial peptides mechanism, efficacy, safety, and other related concern and has helped in creating online database service as well as the future potential of AMPs [ 22 , 153 , 154 ]. Incontestable need for new ways to manage infections and the proven importance of peptides in innate immunity should render the investment worthwhile for human medicine.…”