Interaction of the antimicrobial peptide melimine with bacterial membranes.
Melimine is a novel cationic peptide possessing broad-spectrum antimicrobial activity that is retained when attached to a surface, suggesting that interactions with bacterial membranes may be of primary importance to its activity. The effects of alterations in the environment on the conformation of melimine were investigated using circular dichroism and fluorescence spectra in membrane-mimetic solvents. Furthermore, the interactions of melimine with bacterial membranes of Pseudomonas aeruginosa and Staphylococcus aureus were examined using scanning electron and fluorescence microscopy, and perturbation of membrane integrity was tested by measurement of melimine-mediated diSC(3)-5 dye release from bacterial cells. Melimine has a predominantly random coil conformation that adopts a helical fold when exposed to organic solvents. However, when it is solubilised in micelles of sodium dodecyl sulphate, which are bacterial membrane-mimetic, the alpha-helical content increases to ca. 35-40%. A major effect of melimine was on the integrity of the cytoplasmic membrane both for P. aeruginosa and S. aureus. However, for P. aeruginosa the rapid loss of cytoplasmic membrane integrity correlated directly with loss of cell viability, whilst for S. aureus maximal dye release was obtained at concentrations where there was no significant loss of viability. There have been few studies to date investigating differences in the action of cationic peptides towards Gram-positive and Gram-negative bacteria. Consequently, further investigation of these mechanistic differences may allow more refined targeting of increasingly difficult-to-treat bacterial infections and/or further inform design of novel peptides with improved broad-spectrum activity.
Institute for Eye Research, The University of New South Wales (UNSW), Sydney, NSW 2052, Australia.
R
R
Rasul
N
N
Cole
D
D
Balasubramanian
R
R
Chen
N
N
Kumar
M D P
MD
Willcox
D023181:Antimicrobial Cationic Peptides / Q000378:metabolism* / Q000494:pharmacology*; D002462:Cell Membrane / Q000187:drug effects* / Q000648:ultrastructure; D002942:Circular Dichroism; D005456:Fluorescent Dyes / Q000378:metabolism; D050296:Microbial Viability / Q000187:drug effects; D008855:Microscopy, Electron, Scanning; D008856:Microscopy, Fluorescence; D011550:Pseudomonas aeruginosa / Q000187:drug effects* / Q000648:ultrastructure; D013050:Spectrometry, Fluorescence; D013211:Staphylococcus aureus / Q000187:drug effects* / Q000648:ultrastructure
D023181:Antimicrobial Cationic Peptides; D005456:Fluorescent Dyes; C541912:melimine