Helix stability in barstar peptides.

Abstract

The complex between the ribonuclease barnase and barstar, its intracellular inhibitor, is a very good model for studying protein folding and molecular recognition. We have studied the stability of different peptides that cover the barstar alpha-helix2 involved in the binding to barnase. A linear correlation between the helical amphipathy of these peptides and their inhibitory ability was obtained: the more helically amphipathic, the more the affinity for barnase. We estimated the amount of helix of these peptides in water and in trifluoroethanol by circular dichroism. There is a moderate correlation between the helical amphipathy and the helical content in water, in agreement with previous results that have shown the importance of the hydrophobicity periodicity in the design of peptides. The helical content in trifluoroethanol is related to helical propensity and helical amphipathy, suggesting that the local sequence determines these maximum helicities. The predicted helicity of these peptides, obtained using the algorithm AGADIR [Muñoz, V. & Serrano, L. (1994) Nat. Struct. Biol. 1, 399-409], appears to correlate with their ability to inhibit the activity of barnase in water. The correlation of inhibition constants, helical content in water, and maximum content of helix in trifluoroethanol with helical amphipathy supports the very important role of hydrophobicity pattern in peptide stability.