Escherichia coli cell surface perturbation and disruption induced by antimicrobial peptides BP100 and pepR

This research was originally published in Journal of Biological Chemistry. Carla S. Alves… (et al.). Escherichia coli cell surface perturbation and disruption induced by antimicrobial peptides BP100 and pepR. Journal of Biological Chemistry. 2010. VOL. 285, NO. 36, pp. 27536–27544. © the American Society for Biochemistry and Molecular BiologyThe potential of antimicrobial peptides (AMPs) as an alternative to conventional therapies is well recognized. Insights into the biological and biophysical properties of AMPs are thus key to understanding their mode of action. In this study, the mechanisms adopted by two AMPs in disrupting the Gram-negative Escherichia coli bacterial envelope were explored. BP100 is a short cecropin A-melittin hybrid peptide known to inhibit the growth of phytopathogenic Gram-negative bacteria. pepR, on the other hand, is a novel AMP derived from the dengue virus capsid protein. Both BP100 and pepR were found to inhibit the growth of E. coli at micromolar concentrations. Zeta potential measurements of E. coli incubated with increasing peptide concentrations allowed for the establishment of a correlation between the minimal inhibitory concentration (MIC) of each AMP and membrane surface charge neutralization. While a neutralization-mediated killing mechanism adopted by either AMPis not necessarily implied, the hypothesis that surface neutralization occurs close to MIC values was confirmed. Atomic force microscopy (AFM) was then employed to visualize the structural effect of the interaction of each AMP with the E. coli cell envelope. At their MICs, BP100 and pepR progressively destroyed the bacterial envelope, with extensive damage already occurring 2 h after peptide addition to the bacteria. A similar effect was observed for each AMP in the concentration-dependent studies. At peptide concentrations below MIC values, only minor disruptions of the bacterial surface occurred.This work was supported by the Fundação, para Ciência e Tecnologia, Portugal (Fellowships SFRH/BD/24547/2005, SFRH/BD/24778/2005, and SFRH/BD/39039/2007, to C. S. A., M. N. M., and H. G. F., respectively, and Projects PTDC/QUI/69937/2006 and REEQ/140/BIO/2005)