Positional Effects on Helical Ala-Based Peptides

Helix-coilequilibriumstudiesareimportantforunderstandinghelixformationinproteinfolding, and for helical foldamer design. The quantitative description of a helix using statistical mechanical models is based on experimentally derived helix propensities and the assumption that helix propensity is position- independent. To investigate this assumption, we studied a series of 19-residue Ala-based peptides, to measure the helix propensity for Leu, Phe, and Pff at positions 6, 11, and 16. Circular dichroism spectroscopy revealed that substituting Ala with a given amino acid (Leu, Phe, or Pff) resulted in the following fraction helix trend: KXaa16>KXaa6>KXaa11.HelixpropensitiesforLeu,Phe,andPffatthedifferentpositionswerederived from the CD data. For the same amino acid, helix propensities were similar at positions 6 and 11, but much higher at position 16 (close to the C-terminus). A survey of protein helices revealed that Leu/Phe-Lys (i, i þ 3) sequence patterns frequently occur in two structural patterns involving the helix C-terminus; however, these casesincludealeft-handedconformationresidue.Furthermore,noLeu/Phe-Lysinteractionwasfoundexcept for the Lys-Phe cation-πinteraction in two cases of Phe-Ala-Ala-Lys. The apparent high helix propensity at position 16 may be due to helix capping, adoption of a 3 10-helix near the C-terminus perhaps with Xaa-Lys (i, i þ 3) interactions, or proximity to the peptide chain terminus. Accordingly, helix propensity is generally position-independent except in the presence of alternative structures or in the proximity of either chain terminus. These results should facilitate the design of helical peptides, proteins, and foldamers.