Nature of the charged-group effect on the stability of the C-peptide helix.

Abstract
The residues responsible for the pH-dependent stability of the helix formed by the isolated C-peptide (residues 1-13 of ribonuclease A) were identified by chemical synthesis of analogs and measurement of their helix-forming properties. Each of the residues ionizing between pH 2 and pH 8 was replaced separately by an uncharged residue. Protonation of Glu-2- is responsible for the sharp decrease in helix stability between pH 5 and pH 2, and deprotonation of His-12+ causes a similar decrease between pH 5 and pH 8. Glu-9- is not needed for helix stability. The results cannot be explained by the Zimm-Bragg model and host-guest data for a.sbd.helix formation, which predict that the stability of the C-peptide helix should increase when Glu-2- is protonated or when His-12+ is deprotonated. Histidine+ is a strong helix-breaker in host-guest studies. In proteins, acidic and basic residues tend to occur preferentially near the NH2-terminal end and basic residues near the COOH-terminal end. A possible explanation, based on a helix dipole model was given [Blagdon et Goodman, (1975)]. The results are consistent with the helix dipole model. The distribution of charged residues in protein helices reflects the helix-stabilizing propensity of those residues. Because Glu-9 is not needed for helix stability, a possible Glu-9-.cntdot..cntdot..cntdot.His-12+ salt bridge does not contribute significantly to helix stability. The role of a possible Glu-2-.cntdot..cntdot..cntdot.Arg-10+ salt bridges has not yet been evaluated. A charged-group effect on .alpha.-helix stability in water was also observed in a different peptide system [Ihara, et al, (1982)]; block copolymers containing (Ala)20 and (Glu)20 show partial helix formation at low temperatures, pH 7.5, where the glutamic acid residues are ionized. (Glu)20(Ala)20Phe forms a helix that is markedly more stable than (Ala)20(Glu)20Phe. The results are consistent with a helix dipole model.