Abstract
Conformational studies of nociceptin (NC‐NH~2~), its fully active fragment, NC(1–13)‐NH~2~, and two significantly less potent fragments, NC(1–13)‐OH and NC(1–11)‐OH, were conducted in water and TFE solutions by the employment of circular dichroism, and in DMSO‐d~6~ by 2D NMR spectroscopy in conjunction with theoretical conformational analysis. The conformations of all the peptides studied were calculated taking two approaches. The first assumes multiconformational equilibrium of the peptide studied, which is characterized by a set of conformations (and their statistical weight values) obtained from a global conformational analysis using three methods: the electrostatically driven Monte‐Carlo (EDMC) with the ECEPP/3 force field, the simulated annealing (SA) protocols in the AMBER and CHARMM force fields. The second approach incorporates the interproton distance and dihedral angle constraints into the starting conformation. Calculations were performed using the distance geometry and SA protocol in the CHARMM force field implemented in the X‐PLOR program. The CD experiments indicated that for the active peptides, hydrophobic solvents induced a significantly higher (compared with those remaining) content order, probably a helical structure. Unfortunately, as a result of the conformational flexibility of the peptides, the analysis of conformations obtained with both approaches and different force fields did not allow the selection of any structural elements of the NC peptides that might be connected with their bioactivity. The only common element found in most conformations of the active peptides was a helical character of fragment 8–13, which allowed the side chains of basic amino acid residues to be exposed to the outside of the molecule and probably to interact with the ORL1 receptor. Copyright © 2004 European Peptide Society and John Wiley & Sons, Ltd.