Interactions of Galleria mellonella Proline-Rich Antimicrobial Peptides with Gram-Negative and Gram-Positive Bacteria

Two proline-rich antimicrobial peptides (PrAMPs), named P1 and P2, purified from hemolymph of the greater wax moth Galleria mellonella, were studied for their effects on Gram-negative (Escherichia coli) and Gram-positive (Micrococcus luteus) bacteria. Both peptides decreased the M. luteus bacterial survival rate and caused E. coli bacterial membrane permeabilization. However, in both cases, the P2 peptide was approximately three times more effective than the P1 peptide. Fluorescence microscopy imaging demonstrated binding of both FITC-labeled peptides to E. coli and M. luteus cells. Atomic force microscopy (AFM) and scanning electron microscopy (SEM) imaging of peptide-treated bacteria revealed considerable changes in cell morphology, cell surface topography, and nanomechanical properties. The interactions of the PrAMPs with bacterial cells were also analyzed by FTIR spectroscopy. The P1 peptide action toward E. coli led to partial aggregation of proteins, whereas treatment with P2 resulted in reduced protein aggregation, reflecting differences between both G. mellonella PrAMPs antibacterial action. Moreover, both PrAMPs caused a decrease and an increase in the protein content in relation to lipids on the E. coli and M. luteus cell surface, respectively. The obtained results reflect not only differences between the G. mellonella P1 and P2 peptides but also differences in the cell surface between Gram-negative and Gram-positive bacteria. Both characterized G. mellonella PrAMPs are further representatives of proline-rich peptides with a membrane-permeabilizing antimicrobial mode of action.