Neighbor effect on conformational spaces of alanine residue in azapeptides

The conformational properties of Alanine (Ala) residue have been investigated to understand protein folding and develop force fields. In this work, we examined the neighbor effect on the conformational spaces of Ala residue using model azapeptides, Ac-Ala-azaGly-NHMe (<b>3</b>, <b>AaG</b>), and Ac-azaGly-Ala-NHMe (<b>4, aGA1</b>). Ramachandran energy maps were generated by scanning (φ, ψ) dihedral angles of the Ala residues in models with the fixed dihedral angles (φ = ±90°, ψ = ±0° or ±180°) of azaGly residue using LCgau-BOP and LCgau-BOP + LRD functionals in the gas and water phases. The integral-equation-formalism polarizable continuum model (IEF-PCM) and a solvation model density (SMD) were employed to mimic the solvation effect. The most favorable conformation of Ala residue in azapeptide models is found as the polyproline II (β_P), inverse γ-turn (γ'), β-sheet (β_S), right-handed helix (α_R), or left-handed helix (α_L) depending on the conformation of neighbor azaGly residue in isolated form. Solvation methods exhibit that the Ala residue favors the β_P, δ_R, and α_R conformations regardless of its position in azapeptides <b>3</b> and <b>4</b> in water. Azapeptide <b>5</b>, Ac-azaGly-Ala-NH₂ (<b>aGA2</b>), was synthesized to evaluate the theoretical results. The X-ray structure showed that azaGly residue adopts the polyproline II (β_P) and Ala residue adopts the right-handed helical (α_R) structure in <b>aGA2</b>. The conformational preferences of <b>aGA2</b> and the dimer structure of <b>aGA2</b> based on the X-ray structure were examined to assess the performance of DFT functionals. In addition, the local minima of azapeptide <b>6</b>, Ac-Phe-azaGly-NH₂ (<b>FaG</b>), were compared with the previous experimental results. SMD/LCgau-BOP + LRD methods agreed well with the reported experimental results. The results suggest the importance of weak dispersion interactions, neighbor effect, and solvent influence in the conformational preferences of Ala residue in model azapeptides.