Quantification of the effects of n-π* interactions on the H-bonding properties of amide groups

Abstract

The n-π* interaction is postulated to play a key role in the folding of proteins, especially in proline-rich structures such as collagen, and cooperativity between H-bonding and the n-π* interaction has been proposed. In order to obtain experimental evidence for these cooperative effects, the H-bond acceptor properties of secondary amides with and without the capacity to form an intramolecular n-π* interaction were measured. UV-vis absorption and ¹³C NMR titrations were used to investigate the intermolecular H-bonded complexes formed with 2-methyl-4-nitro-phenol and perfluoro-tert-butanol, and hence quantify the H-bond acceptor properties of the amide carbonyl oxygens. For an N-acylproline derivative, the presence of an intramolecular n-π* interaction between two amide groups was confirmed by X-ray crystallography, but the solution titrations show that associated changes in the H-bond acceptor strength of the amide carbonyl oxygen group are negligible. The free energy contribution due to cooperativity between the intramolecular n-π* interaction and the intermolecular H-bond was found to be within the error of the experiment (<1 kJ mol⁻¹). The results suggest that any contributions to the thermodynamic stability of folded proteins due to such cooperativity are small.