Energetics, structures, vibrational frequencies, vibrational absorption, vibrational circular dichroism and Raman intensities of Leu-enkephalin

Here we present several low energy conformers of Leu-enkephalin (LeuE) calculated with the density functional theory using the Becke 3LYP hybrid functional and the 6-31G* basis set. The structures, conformational energies, vibrational frequencies, vibrational absorption (VA) intensities, vibrational circular dichroism (VCD) intensities and Raman scattering intensities are reported for the conformers of LeuE which are expected to be populated at room temperature. The species of LeuE-present in non-polar solvents is the neutral non-ionic species with the NH2 and CO2H groups, in contrast to the zwitterionic neutral species with the NH3+ and CO2- groups which predominates in aqueous solution and in the crystal. All of our attempts to find the zwitterionic species in the isolated state failed, with the result that a hydrogen atom from the positively charged N-terminus ammonium group transferred either to one of the oxygens of the carboxylate group of the C-terminus or to the oxygen of the amide group of one of the other residues. Hence we conclude that the zwitterionic species of LeuE is not stable in the isolated state. Spectral simulations of the species expected to be found in the isolated state can be compared to the measured VA, VCD and Raman spectra of LeuE in non-polar solvents to identify which conformer or conformers of LeuE are present in these media. Characteristic features in the VCD spectra are more sensitive to conformational changes than those in either the VA or Raman spectra, similar to the characteristic features in electronic circular dichroism spectra with respect to those in the UV-vis electronic absorption spectra. Finally, we have also attempted to stabilize the zwitterionic species by treating the aqueous environment by using a continuum solvent approach, the Onsager model. Here we found that the zwitterionic species is now stable. The neutral species in an aqueous environment was also modelled by the continuum solvent approaches to determine the relative stability of these two species in this new aqueous environment. Here the relative energy of the zwitterionic species is higher than neutral species, in contradiction to experiment. Hence the use of explicit water molecules plus either this or another continuum model to treat the bulk water environment is necessary to make the zwitterionic species more stable than the neutral species. We are pursuing explicit water molecules to treat LeuE in this environment.