Conformation induction in melanotropic peptides by trifluoroethanol: fluorescence and circular dichroism study.

Conformation induction in the two related peptides, alpha-melanocyte stimulating hormone (alpha-MSH) and delta-melanocyte stimulating hormone (delta-MSH), have been studied in solvent media containing varying percentages of the membrane-mimetic solvent 2,2,2-trifluoroethanol (TFE) using fluorescence and circular dichroism (CD) spectroscopy. Singular value decomposition (SVD) analysis of the CD spectra at different TFE concentrations showed that these spectra can be described as linear combinations of only two distinct basis spectra, corresponding to the peptides in the random-coil and 'folded' conformations. For alpha-MSH the spectrum of the folded state is very similar to the standard spectrum of the alpha-helix, while that for delta-MSH has partial resemblance to the helical spectrum. Fitting the data on ellipticity (at 222 nm) as a function of TFE volume fraction to an equation based on a two-state model describing TFE-induced conformation induction in the peptides gave values of (1.1 +/- 0.4) and (4.2 +/- 0.5) kcal mol-1 for alpha-MSH and delta-MSH, respectively, for the free energy of equilibrium between the helix and coil forms in water. Measurement of fluorescence emission parameters (emission maximum, quantum yield, steady-state anisotropy and mean excited-state lifetime) indicated that the microenvironment around the single tryptophan residues of both peptides changes in like manner with increasing concentration of TFE in the solvent. The similarity of fluorescence behaviour of the peptides suggests that their Trp fluorophores do not participate in secondary structure formation in TFE.