Open AccessKinetics of membrane lysis by custom lytic peptides and peptide orientations in membrane
Author(s) -
Chen H. M.,
Clayton A. H. A.,
Wang W.,
Sawyer W. H.
Publication year - 2001
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1046/j.1432-1327.2001.02039.x
Subject(s) - liposome , lysis , phospholipid , chemistry , membrane , peptide , bilayer , lipid bilayer , circular dichroism , biophysics , phosphatidic acid , melittin , kinetics , biochemistry , biology , physics , quantum mechanics
To aid the development of custom peptide antibiotics, a kinetic study of membrane lysis by cecropin B (CB) and its analogs, cecropin B1 (CB1) and cecropin B3 (CB3) was carried out to determine the mechanism by which these peptides disrupt the bilayer structure of liposomes of defined composition. Disruption of the phospholipid bilayer was determined by a fluorescence assay involving the use of dithionite to quench the fluorescence of lipids labeled with N ‐7‐nitro‐2,1,3‐benzoxadiazol‐4‐yl. Lytic peptides caused the disruption of liposomes to occur in two kinetic steps. For liposomes composed of mixtures of phosphatidylcholine and phosphatidic acid, the time constants for each kinetic step were shorter for CB and CB1 than for CB3. Oriented circular dichroism experiments showed that the peptides could exist in at least two different membrane‐associated states that differed primarily in the orientation of the helical segments with respect to the bilayer surface. The results are discussed in terms of kinetic mechanisms of membrane lysis. The mode of actions of these peptides used for the interpretation of their kinetic mechanisms were supported by surface plasmon resonance experiments including or excluding the pore‐forming activities.