Interaction of fluorescently labeled pardaxin and its analogues with lipid bilayers.
Fluorescence measurements were used to monitor the interaction of the neurotoxin pardaxin and its analogues with membranes. Eight peptides were selectively labeled with the fluorophore 7-nitrobenz-2-oxa-1,3-diazole-4-yl, either at their N-terminal or at their C-terminal. No detectable changes in membrane permeability or hemolytic activity were observed upon modification. Upon the titration of solutions containing the different peptides with small unilamellar vesicles, the fluorescent emission spectra of 7-nitrobenz-2-oxa-1,3-diazole-4-yl-labeled pardaxin and its analogues, but not those of control peptides, displayed blue shifts in addition to enhanced intensities upon relocation of the probe to a more apolar environment. The results revealed that the N terminus of pardaxin is buried within the lipid bilayer while the C terminus is located at the bilayer's surface. Binding isotherms were obtained from the observed increases in the fluorescence emission yields, from which surface partition constants, in the range of 10(4) M-1, were in turn derived. The existence of an aggregation process was suggested by the shape of the binding isotherms. Furthermore, the results show good correlation between the incidence of aggregation and the ability of the different analogues to induce the release of relatively large molecules from vesicles. As such, our results suggest that the mechanism of pore formation employed by pardaxin and its analogues could be described by the "barrel stave" model.
Rapaport
D
D
Department of Membrane Research and Biophysics, Weizmann Institute of Science, Rehovot, Israel.
Shai
Y
Y
D009327:4-Chloro-7-nitrobenzofurazan; D000595:Amino Acid Sequence; D004058:Diffusion; D005397:Fish Venoms / Q000737:chemistry*; D007700:Kinetics; D008051:Lipid Bilayers; D008969:Molecular Sequence Data; D009498:Neurotoxins / Q000737:chemistry; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry*; D013050:Spectrometry, Fluorescence; D013329:Structure-Activity Relationship
D005397:Fish Venoms; D008051:Lipid Bilayers; D009498:Neurotoxins; D010455:Peptides; C017603:pardaxin; D009327:4-Chloro-7-nitrobenzofurazan