Tryptophan fluorescence studies of melanotropins in the amphiphile-water interface of reversed micelles.
We report studies on the interaction of alpha-melanocyte stimulating hormone (alpha-MSH) and a synthetic analogue (MSH-I) with reverse micelles prepared from the amphiphilic sodium bis(2-ethylhexyl)sulfosuccinate in isooctane. The tripeptide lysyl-tryptophyl-lysine and the isolated amino acid tryptophan were also investigated as simpler compounds interacting with the micelles. Tryptophan fluorescence parameters (spectral position of emission band, anisotropy, and lifetime decay) demonstrated that in the presence of reverse micelles the environment around the fluorophore is less polar and more rigid than bulk water. Those parameters are sensitive to the changes induced in the micelles by the presence of water. In large micelles having a water/ amphiphile molar ratio above 10, the modifications detected by fluorescence are small and the location of the fluorophore is not affected by a further increase in the concentration of the bulk water. The results, with additional support from quenching experiments, indicated that the different compounds occupy different positions in the large reverse micelles, but in any case they are in the interface region, without dispersing into the bulk water. From decay associated spectra, conformations were identified showing different degrees of tryptophan exposition to polar and nonpolar local environments. The conformation related to the long lifetime has its tryptophan more exposed to water while that associated to the intermediate lifetime has that residue stabilized in nonpolar media. The native hormone alpha-MSH and the analogue MSH-I present similar conformations in dry micelles. However, in buffer and in the large hydrated micelles, differences in conformations are evident, and could be related to the different physiological activity of the peptides.
Souto
A L
AL
Instituto de Física, Universidade de São Paulo, SP, Brazil.
Ito
A S
AS
D005454:Fluorescence Polarization; D008823:Micelles; D009842:Oligopeptides / Q000737:chemistry; D011487:Protein Conformation; D011789:Quantum Theory; D013050:Spectrometry, Fluorescence; D014364:Tryptophan / Q000737:chemistry*; D000521:alpha-MSH / Q000031:analogs & derivatives / Q000737:chemistry*
D008823:Micelles; D009842:Oligopeptides; C029535:lysyl-tryptophyl-alpha-lysine; D000521:alpha-MSH; D014364:Tryptophan