Abstract
A series of 14 residue amphipathic α‐helical peptides, in which the sidechains of glutamic acid and lysine have been covalently joined, was synthesized in order to determine the effect of spacing, position and orientation of these lactam bridges. It was found that although an (i, i+3) spacing would position the lactam bridge on the same face of the helix, these lactams with 18‐member rings were actually helix‐destabilizing regardless of position or location. On the other hand, (i, i+4) lactams with 21‐member rings were helix‐stabilizing but this was dependent on orientation. Glutamic acid‐lysine lactams increased the helical content of the peptide when compared with their linear homologue in benign conditions (50 mM KH~2~PO~4~, 100 mM KCl, pH 7). Two Glu‐Lys (i, i+4) lactams located at the N‐ and C‐termini gave rise to a peptide with greater than 99% helical content in benign conditions. Peptides with Lys‐Glu oriented lactams were random structures in benign conditions but in the presence of 50% TFE could be induced into a helical conformation. The stability of the single‐stranded α‐helices, as measured by thermal denaturations in 25% TFE indicated that Glu‐Lys oriented lactam bridges stabilized the helical conformation relative to the linear unbridged peptide. One Glu‐Lys lactam in the middle of the peptide was more effective at stabilizing helical structure than two Glu‐Lys lactams positioned one at each end of the molecule. The lactams with the Lys‐Glu orientation were destabilizing relative to the unbridged peptide. This study demonstrates that correct orientation and position of a lactam bridge is critical in order to design peptides with high helical content in aqueous media.