Effect of salt and membrane fluidity on fluorophore motions of a gramicidin C derivative.
We have used fluorescence spectroscopy to investigate the effect of salt and membrane fluidity on the rotational motion of a 5-(dimethylamino)naphthalene-1-sulfonyl (dansyl) derivative of gramicidin C (dansyl-gC) in dimyristoylphosphatidylcholine bilayers, under conditions where the peptide is a formyl-NH to formyl-NH terminal dimer, and in octyl glucoside micelles, where the peptide is an intertwined helical dimer. Energy-transfer experiments showed no changes in either conformation or dimer aggregation under the conditions explored here (15-40 degrees C, 1-350 bar, 0-0.33 M Mg2+, and 0-1 M Na+). The addition of permeable (Na+) or nonpermeable (Mg2+) ions did not affect the temperature or pressure behavior of dansyl rotation. However, fluorescence lifetime measurements indicated an increase in solvent accessibility in the presence of sodium. In bilayers, the temperature dependence of the fluorescence polarization and lifetime shows strong interactions between the dansyl residue and the peptide, and at no time did the dansyl motions become solvent controlled as has been observed for aqueous solvent peptides [Scarlata, S. F., Rholam, M., & Weber, G. (1984) Biochemistry 23, 6789]. In micelles, the change in rotational motion with temperature followed solvent expansion, showing that in this case the dansyl residue does not associate extensively with the peptide. Our results indicate that because of the extensive coupling between the dansyl residue and the rest of the peptide, membrane fluidity does not play a major role in controlling side-chain motions.
Teng
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Department of Medicine and Physiology and Biophysics, Cornell University Medical College, New York, New York 10021.
Koeppe
R E
RE
Scarlata
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SF
D000595:Amino Acid Sequence; D003619:Dansyl Compounds; D004735:Energy Transfer; D006096:Gramicidin; D007700:Kinetics; D008274:Magnesium / Q000494:pharmacology*; D008560:Membrane Fluidity; D008954:Models, Biological; D008969:Molecular Sequence Data; D011312:Pressure; D011487:Protein Conformation; D012964:Sodium / Q000494:pharmacology*; D013050:Spectrometry, Fluorescence; D013816:Thermodynamics; D014783:Viscosity
D003619:Dansyl Compounds; D006096:Gramicidin; D012964:Sodium; D008274:Magnesium