Ib-AMP1

Continual occurrence of foodborne outbreaks, along with the increase in antibiotic resistance which burdens clinical treatments, has urged scientists to search for other potential promising antimicrobial agents. Antimicrobial peptides are emerging as one of the potential alternatives. The mode of action of a given AMP is critical and essential for future application; however, it is still not completely known for many of these compounds. Ib-AMP1 is a plant-derived AMP, purified from seeds of Impatiens balsamina and has been shown to exert antibacterial and antifungal activity at the micromolar level. A study had shown that the therapeutic index of Ib-AMP1 against eight human pathogens is 23.5. The objective of the present study was to determine the in vivo mode of action of Ib-AMP1 against Escherichia coli O157:H7. A concentration-dependent effect of Ib-AMP1 on the E. coli O157:H7 cell membrane occurred. Ib-AMP1 treatments resulted in efflux of K(+) and ATP, suggesting pores of sufficient size to allow efflux of large molecules. Ib-AMP1 at sublethal concentrations exerts a greater effect at the intracellular level. In contrast, Ib-AMP1 at a lethal concentration permeabilizes cell membranes and may directly or indirectly inhibit intracellular macromolecule synthesis. Collectively, results of this study suggest Ib-AMP1 is bactericidal interfering within outer and inner membrane integrity permitting efflux of ATP and interfering with intracellular biosynthesis of DNA, RNA and protein.

Due to an increase in the occurrence of multi drug resistant microorganisms a need for the development of alternative drugs comes in light. This alternative drug should be such that the microorganisms should not be able to develop resistance against them easily. Antimicrobial peptides are the most potential candidates to be developed as alternative drug. In the present study the three toxins ETA, ETB and PVL of Staphylococcus aureus were docked with four antimicrobial peptides, Ib-AMP1, JCPep7, Snakin2, Sesquin, derived from plants. The docking studies predict that Ib-AMP1 shows significant interactions with all these three toxins. Hence, further studies can be carried out for developing Ib-AMP1 as an alternative drug against the toxins of Staphylococcus aureus.

Ib-AMP1 is a 20-residue disulfide-linked beta-sheet antimicrobial peptide found in the seeds of Impatiens balsamina. In order to investigate the effects of the 2 disulfide bonds on the antimicrobial specificity, to determine the mechanism of antimicrobial action of Ib-AMP1 and to develop novel cell-selective antimicrobial peptides with improved antimicrobial specificity as compared to wild-type Ib-AMP1, we synthesized a disulfide-removed linear analog of Ib-AMP1 with L-Pro, D-Pro or peptoid residues (Nala and Nlys) at the central position of the molecule. All linear analogs displayed a 3.7-4.8-fold higher antimicrobial specificity than wild-type Ib-AMP1, indicating that the disulfide bonds of Ib-AMP1 analogs are not essential for its antimicrobial specificity. Circular dichroism spectra revealed that the peptoid residues, as well as the proline at the central position of disulfide bond-removed Ib-AMP1 analogs, induce a beta-turn structure in a negatively charged bacterial membrane-mimicking environment. Ib-AMP1 was not effective in depolarizing the cytoplasmic membranes of Staphylococcus aureus and showed almost no leakage of calcein from negatively charged bacterial membranes mimicking lipid vesicles. In contrast, all linear analogs caused very weak dye leakage from negatively charged vesicles, but they almost completely depolarized the membrane potential of S. aureus cells. Collectively, our results suggest that the target of Ib-AMP1 may not be the cytoplasmic membranes of bacteria but their intracellular components. All linear analogs exhibit lethality due to their ability to form small channels that permit the transit of ions or protons and not molecules as large as calcein, and not by disrupting membranes.