title: Binding and permeabilization of model membranes by amphipathic peptides.
abstract: Measurement of binding and activity of antimicrobial and cytolytic amphipathic peptides on membranes is essential to understanding their function and cell specificity. The use of model systems has provided a wealth of information on the interactions of amphipathic peptides with membranes. Binding of peptides to membranes can be monitored by measuring Förster resonance energy transfer from a Trp residue on the peptide to a lipid fluorophore incorporated in the membrane. Especially for peptides that perturb or disrupt the membrane, it is advantageous to perform these measurements as a function of time, rather than in steady state. The activity of these amphipathic peptides toward model membranes is usually measured by dye efflux kinetics. One of those methods, based on self-quenching of carboxyfluorescein, is described here, together with a discussion of caveats and pitfalls of the corresponding analysis and interpretation.
authors.affiliation: Department of Chemistry and Biochemistry, University of North Carolina Wilmington, Wilmington, NC, USA.
authors.forename: Paulo F
authors.initials: PF
authors.lastname: Almeida
authors.forename: Antje
authors.initials: A
authors.lastname: Pokorny
mesh_terms: D023181:Antimicrobial Cationic Peptides / Q000378:metabolism*; D002463:Cell Membrane Permeability; D005452:Fluoresceins / Q000378:metabolism; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D007700:Kinetics; D011485:Protein Binding; D053835:Unilamellar Liposomes / Q000378:metabolism*
chemical_list: D023181:Antimicrobial Cationic Peptides; D005452:Fluoresceins; D053835:Unilamellar Liposomes; C024098:6-carboxyfluorescein