title: Slow insertion kinetics during interaction of a model antimicrobial peptide with unilamellar phospholipid vesicles.
abstract: The mechanism of interaction between a model antimicrobial peptide and phospholipid unilamellar vesicle membranes was studied using fluorescence spectroscopy, fluorescence lifetime measurements, and light scattering. The peptide, a mellitin mutant, was labeled at position K14 with the polarity-sensitive probe AlexaFluor 430. The kinetics of the interaction of this derivative with various concentrations of 1,2-dipalmitoyl-sn-glycero-3-phosphatidylcholine (DPPC) vesicles was examined. Our work unveiled two novel aspects of peptide-lipid interactions. First, the AB plot or phasor analysis of the fluorescence lifetime studies revealed at least three different peptide states, the population of which depended on the lipid to peptide (L:P) concentration ratio. Second, complex fluorescence kinetics were observed over extended time-scales from 30 s to 2 h. The extended kinetics was only observed at particular lipid concentrations (L:P ratios 20:1 and 10:1) and not at others (30, 40, 50 and 100:1 L:P ratio). Analysis of the complex kinetics revealed several intermediates. We assign these to distinct states of the peptide formed during helix insertion into the vesicle membrane that are intermediate to lytic pore formation.
authors.affiliation: School of Chemistry, University of Melbourne, Parkville, Victoria 3010, Australia.
authors.forename: Zubaidah
authors.initials: Z
authors.lastname: Ningsih
authors.forename: Mohammed Akhter
authors.initials: MA
authors.lastname: Hossain
authors.forename: John D
authors.initials: JD
authors.lastname: Wade
authors.forename: Andrew H A
authors.initials: AH
authors.lastname: Clayton
authors.forename: Michelle L
authors.initials: ML
authors.lastname: Gee
mesh_terms: D015060:1,2-Dipalmitoylphosphatidylcholine / Q000378:metabolism*; D000595:Amino Acid Sequence; D005456:Fluorescent Dyes / Q000378:metabolism; D007700:Kinetics; D008555:Melitten / Q000737:chemistry / Q000378:metabolism*; D008969:Molecular Sequence Data; D011485:Protein Binding; D053835:Unilamellar Liposomes / Q000737:chemistry / Q000378:metabolism*
chemical_list: D005456:Fluorescent Dyes; D053835:Unilamellar Liposomes; D008555:Melitten; D015060:1,2-Dipalmitoylphosphatidylcholine