A Meccano set approach of joining trpzip a water soluble β-hairpin peptide with a didehydrophenylalanine containing hydrophobic helical peptide

Abstract

A 16 residues long, water soluble, monomeric β -hairpin peptide 'trpzip' (Cochran et al. Proc. Natl. Acad. Sci. U.S.A., 98 (2001), 5578), stabilized by tryptophan zipper has been linked via a tetraglycyl linker to a hydrophobic didehydrophenylalnine (ΔF) containing helical octapeptide. Circular dichroism studies of this 28 residues long peptide, 'trpzipalpha' (Ac-GEWTWDDATKTWTWTE-GGGG-ΔFALΔFALΔFA-NH₂) in water have revealed the presence of both the β-hairpin and the helical conformations. This is the first instance where a ΔF containing peptide has been found to display a helical fold in water. The fluorescence emission wavelengths of tryptophan in Ac-G-W-G-NH₂, trpzip and trpzipalpha were 341.5, 332.8 and 332.6 nm, respectively. The fluorescence quantum yield of trpzip was 2.6-fold higher than trpzipalpha suggesting that proximal interactions between the β-hairpin and the helix caused the quenching of tryptophan fluorescence in the former by the ΔFs in the latter. The molar ellipticity of the far UV couplet characteristic of trpzip was reduced in trpzipalpha and the CD based thermal melting temperatures at 228 nm were 62 °C (trpzip) and 57 °C (trpzipalpha). A concentration-dependent variable temperature CD study in water showed that in trpzipalpha, increasing temperature is detrimental to the β-hairpin, but it augments the helical motif, perhaps by intermolecular oligomerization. Our results show that in water, trpzipalpha exhibits long-range interactions between two different secondary structures. In contrast to trpzip, trpzipalpha has shown a greater tendency to oligomerize in water.