Interaction of model class A1, class A2, and class Y amphipathic helical peptides with membranes.

To test the hypothesis that differences in the lipid affinity of exchangeable apolipoproteins are due to the presence of different classes of amphipathic alpha-helical motifs which differ primarily in the distribution of charged amino acid residues, we designed and synthesized model peptides mimicking class A1, class A2, and class Y amphipathic helices present in these apolipoproteins. Both class A1 and class A2 helices have positive residues at the polar-nonpolar interface and negative residues at the center of the polar face. However, clustering of positive and negative residues is less exact in class A1 compared to class A2 helices. The class Y helices have two negative residue clusters on the polar face separating the two arms and the base of the Y motif formed by three positive residue clusters. The lipid affinities of three 18 residue model peptides representing these classes, Ac-18A1-NH2 (Ac-ELLEKWAEKLAALKEALK-NH2), Ac-18A2-NH2 (Ac-ELLEKWKEALAALAEKLK-NH2), and Ac-18Y-NH2 (Ac-ELLKAWKEALEALKEKLA-NH2), were determined by right-angle light scattering, circular dichroism spectroscopy, differential scanning calorimetry, and fluorescence spectroscopy. The observed rank order of lipid affinity of these three peptides is: Ac-18A2-NH2 > Ac-18Y-NH2 > Ac-18A1-NH2. This order is consistent with the known lipid affinity of exchangeable apolipoproteins containing class A1, class A2, and class Y helices (class A2 > class Y > class A1). Results of this study illustrate the important role of interfacial lysine residues in modulating the lipid affinity of amphipathic helices and suggest that the effect of interfacial lysine residues in increasing lipid affinity is additive. We propose that interfacial lysine residues, in addition to widening the hydrophobic face because of snorkeling, also help anchor the amphipathic helix in the lipid bilayer.