First Step Toward the Quantitative Identification of Peptide 310-Helix Conformation with NMR Spectroscopy: NMR and X-ray Diffraction Structural Analysis of a Fully-Developed 310-Helical Peptide Standard

Abstract

We have synthesized by solution methods and fully characterized the Nα-blocked heptapeptide methylamide mBrBz-[L-Iva-L-(αMe)Val]2-L-(αMe)Phe-L-(αMe)Val-L-Iva-NHMe, fully based on conformationally constrained Cα-methylated α-amino acids. An X-ray diffraction investigation of the Nα-benzyloxycarbonylated analogue showed that in the crystal state both independent molecules (A and B) in the asymmetric unit of the peptide adopt a fully developed, regular, right-handed 310-helical structure, although molecule A would be slightly distorted at the C-terminal residue. Solution conformational analysis on the mBrBz-blocked peptide was carried out in CDCl3 by means of NMR spectroscopy. For structure determination we performed restrained molecular dynamics simulations in CDCl3 based on a search of the conformational space derived from a simulated annealing strategy. For this peptide the NMR observables can be described by a single backbone conformation, more specifically a rigid 310-helix spanning the amino acid seque...