Solvent dependence of the rotational diffusion of TOAC-spin-labeled alamethicin

Three derivatives of the hydrophobic, channel-forming peptaibiotic alamethicin (F50/5) have been synthesized, the original Aib residue at position 1, 8, or 16 being replaced with the spin-labeled amino acid TOAC (=2,2,6,6-tetramethylpiperidin-1-oxyl-4-amino-4-carboxylic acid). Electron-paramagnetic-resonance (EPR) spectroscopy was used to characterize the rotational diffusion of these compounds in five isotropic solvents of differing viscosity and polarity, including MeOH, EtOH, PrOH, i-PrOH, and hexanol (HxOH). In MeOH, the labeled alamethicins were found to rotate anisotropically as a monomer (axial ratio a/b=3). In aliphatic alcohols of increasing viscosity (and hydrophobicity), the rotational correlation times progressively increased. Even in HxOH, the (fivefold) increase in correlation time was no greater than the increase in viscosity. We conclude that TOAC-labeled alamethicins remain monomeric in these solvents of relatively high polarity.