Membrane interactions of the sodium channel S4 segment and its fluorescently-labeled analogues.
A 24-amino acid peptide corresponding to the S4 segment of the sodium channel was synthesized. In order to perform fluorescence energy transfer measurements and to monitor the interaction of the peptide with lipid vesicles, the peptide was selectively labeled with fluorescence probes at either its N- or C-terminal amino acids. The fluorescent emission spectra of 7-nitrobenz-2-oxa-1,3-diazol-4- yl-(NBD-)labeled analogues displayed blue shifts upon binding to small unilamellar vesicles (SUV), reflecting the relocation of the fluorescent probe to an environment of increased apolarity. The results revealed that both the N- and C-terminus of the S4 segment are located within the lipid bilayer. Titration of solutions containing NBD-labeled peptides with SUV was used to generate binding isotherms, from which surface partition constants, in the range of 10(4) M-1, were derived. The shape of the binding isotherms as well as fluorescence energy transfer measurements suggest that aggregation of peptide monomers within the membrane readily occurs in acidic but not in zwitterionic vesicles. Furthermore, the results provide good correlation between the incidence of aggregation in PC/PS vesicles and the ability of the peptides to permeate the vesicle's membrane. However, a transmembrane diffusion potential had no detectable effect on the location of the peptide within the lipid bilayer or on its aggregation state.(ABSTRACT TRUNCATED AT 250 WORDS)
Rapaport
D
D
Department of Membrane Research and Biophysics, Weizmann Institute of Science, Rehovot, Israel.
Danin
M
M
Gazit
E
E
Shai
Y
Y
D000595:Amino Acid Sequence; D004058:Diffusion; D004735:Energy Transfer; D005452:Fluoresceins / Q000737:chemistry; D008051:Lipid Bilayers / Q000737:chemistry; D008081:Liposomes; D008560:Membrane Fluidity; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry; D010539:Permeability; D015222:Sodium Channels / Q000737:chemistry*; D012995:Solubility; D013050:Spectrometry, Fluorescence; D014634:Valinomycin / Q000737:chemistry
D005452:Fluoresceins; D008051:Lipid Bilayers; D008081:Liposomes; D010446:Peptide Fragments; D015222:Sodium Channels; D014634:Valinomycin; C007740:fluorexon