However, there was again no information about the I/L differentiation. Edman degradation suggested a 13 amino acid sequence as F-D-I-M-G-L-I-K-K-V-A-G-A, and so,

the C-terminal I/L was still not determined. Finally, it was determined by the solid-phase synthesis of both the 14I and 14L peptides and their HPLC behavior was compared to the natural peptide. As a consequence, the 14L peptide was found to be identical to the natural one, and GSK126 clinical trial therefore, the sequence was unambiguously determined as F-D-I-M-G-L-I-K-K-V-A-G-A-L-NH2. Similarly, eumenitin-F and eumenine mastoparan-EF (EMP-EF) were purified from the extracts of E. fraterculus ( Fig. 1B), and in the same manner, the sequences were determined to be L-N-L-K-G-L-F-K-K-V-A-S-L-L-T and F-D-V-M-G-I-I-K-K-I-A-S-A-L-NH2, respectively.

The chemical features of these new peptides, rich in hydrophobic and basic amino acids with no disulfide bond, are characteristic of linear cationic cytolytic peptides ( Kuhn-Nentwig, 2003), in particular, eumenitin-R and eumenitin-F, are highly homologous to eumenitin, whereas the other two, EMP-ER and EMP-EF, are similar to EMP-AF, thus can be classified as mastoparans ( Fig. 2, Murata et al., 2009). This class of peptides has been known to adopt an amphipathic α-helical conformation, showing an amphiphilic character under appropriate TGF-beta tumor conditions ( Wakamatsu et al., 1992, Hori et al., 2001, Sforça et al.,

2004 and Todokoro et al., 2006). The amphipaticity of peptides has been considered essential for their biological activities (Wimley, 2010). In fact, if the helical wheel projections of these peptide sequences were drawn, they show that amphipathic α-helical conformations could be possible as depicted in Fig. 3. Based on this view, all the hydrophilic amino acid residues, S, T, N and K, are located on one side, whereas the hydrophobic amino acid residues, I, L and V are on the other side of the helix. The Eumenine wasp venom peptides as Liothyronine Sodium well as mastoparan peptides are known to undergo a conformational change from a random coil to helical upon binding to lipid bilayers or in membrane mimetic environments (Park et al., 1995; Santos Cabrera et al., 2004 and Konno et al., 2006). The α-helix content of these short chain peptides is directly related to favorable electrostatic interactions and the burial of the backbone into a more hydrophobic region. Fig. 4 shows the CD spectra of eumenitin-R, eumenitin-F, EMP-ER and EMP-EF obtained in different environments, to evaluate the relative importance of the electrostatic and hydrophobic contributions to the observed ellipticity.