title: The role of electrostatic interactions in the membrane binding of melittin.
abstract: The binding of melittin and the C-terminally truncated analogue of melittin (21Q) to a range of phospholipid bilayers was studied using surface plasmon resonance (SPR). The phospholipid model membranes included zwitterionic dimyristylphosphatidylcholine (DMPC) and dimyristylphosphatidylethanolamine (DMPE), together with mixtures DMPC/dimyristylphosphatidylglycerol (DMPG), DMPC/DMPG/cholesterol and DMPE/DMPG. Melittin bound rapidly to all membrane mixtures, whereas 21Q, which has a reduced charge, bound much more slowly on the DMPC and DMPC/DMPG mixtures reflecting the role of the initial electrostatic interaction. The loss of the cationic residues also significantly decreased the binding of 21Q with DMPC/DMPG/Cholesterol, DMPE and DMPE/DMPG. The role of electrostatics was also highlighted with NaCl in the buffer, which affected the way melittin bound to the different membranes, causing a more uniform, concentration dependant increase in response. The biosensor results were correlated with the conformation of the peptides determined by circular dichroism analysis, which indicated that high α-helicity was associated with high binding affinity. Overall, the results demonstrate that the positively charged residues at the C-terminus of melittin play an essential role in membrane binding, that modulation of peptide charge influences selectivity of binding to different phospholipids and that manipulation of the cationic regions of antimicrobial peptides can be used to modulate membrane selectivity.
authors.affiliation: Department of Biochemistry & Molecular Biology, Monash University, Clayton, Victoria 3800, Australia.
authors.forename: Kristopher
authors.initials: K
authors.lastname: Hall
authors.forename: Tzong-Hsien
authors.initials: TH
authors.lastname: Lee
authors.forename: Marie-Isabel
authors.initials: MI
authors.lastname: Aguilar
mesh_terms: D000595:Amino Acid Sequence; D002942:Circular Dichroism / Q000379:methods; D008051:Lipid Bilayers / Q000737:chemistry* / Q000378:metabolism*; D008555:Melitten / Q000737:chemistry* / Q000378:metabolism*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011485:Protein Binding; D017433:Protein Structure, Secondary; D012965:Sodium Chloride / Q000737:chemistry; D055672:Static Electricity; D020349:Surface Plasmon Resonance
chemical_list: D008051:Lipid Bilayers; D008555:Melitten; D012965:Sodium Chloride